Antioxidant Potential of Propolis, Bee Pollen, and Royal Jelly: Possible Medical Application
Joanna Kocot 乔安娜 · 科科特,1 Małgorzata Kiełczykowska 马格扎塔 · 基奇科沃斯卡,1Dorota Luchowska-Kocot,1 Jacek Kurzepa 亚切克 · 库尔泽帕,1 and 及Irena Musik 伊雷纳音乐1Show more展示更多Academic Editor: 学术编辑:Kota V. Ramana 哥打 v. 拉马纳Received 收到26 Jan 2018 二零一八年一月二十六日Revised 修订本25 Mar 2018 二零一八年三月二十五日Accepted 接受02 Apr 2018 二零一八年四月二日Published 出版02 May 2018 二零一八年五月二日
Honeybees products comprise of numerous substances, including propolis, bee pollen, and royal jelly, which have long been known for their medicinal and health-promoting properties. Their wide biological effects have been known and used since antiquity. Bee products are considered to be a potential source of natural antioxidants such as flavonoids, phenolic acids, or terpenoids. Nowadays, the still growing concern in natural substances capable of counteracting the effects of oxidative stress underlying the pathogenesis of numerous diseases, such as neurodegenerative disorders, cancer, diabetes, and atherosclerosis, as well as negative effects of different harmful factors and drugs, is being observed. Having regarded the importance of acquiring drugs from natural sources, this review is aimed at updating the current state of knowledge of antioxidant capacity of selected bee products, namely, propolis, bee pollen, and royal jelly, and of their potential antioxidant-related therapeutic applications. Moreover, the particular attention has been attributed to the understanding of the mechanisms underlying antioxidant properties of bee products. The influence of bee species, plant origin, geographic location, and seasonality as well as type of extraction solutions on the composition of bee products extracts were also discussed.
Bee products like propolis, bee wax, pollen, royal jelly, as well as honey had been known and used even in antiquity and the Middle Ages. For example, in ancient China, bee pollen was applied as a cosmetic agent contributing to skin whitening. At present, these substances are applied in a branch of complementary and alternative medicine—apitherapy. Moreover, the interest in their use as agents in the cure of cancers, neurodegenerative, cardiovascular, and gastrointestinal tract diseases as well as the treatment of wounds and burns has still been growing [1–10].
Bee products are considered to be a potential source of natural antioxidants capable of counteracting the effects of oxidative stress underlying the pathogenesis of numerous diseases.
In general, the compounds possessing phenolic character, which belong to substances expressing ability to scavenge free radicals, are mainly responsible for bee products’ antioxidant capacity [11–14]. They comprise of two main groups of compounds—flavonoids and phenolic acids .
Flavonoids are plant derivatives of polyphenolic structure comprising several subgroups like flavones, flavonols, flavanones flavanonols, flavanols (catechins), anthocyanins, and chalcones, as well as isoflavones and neoflavonoids. The best known subgroups are the compounds containing benzo-γ-pyrone skeleton. Flavonoids often occur in the form of glycosides, in which they play a role of aglycones connected by a glycosidic bond with a carbohydrate group [15–17]. The presence of phenol groups in the molecules of flavonoids imparts them the antiradical activity all the more because the radicals formed during scavenging are resonance stabilized . The examples of flavonoids and their glycosides found in bee products are presented in Figure 1.
黄酮类化合物是多酚结构的植物衍生物，包括黄酮、黄酮醇、黄酮醇、黄酮醇(儿茶素)、花青素、查尔酮、异黄酮和新黄酮等几个亚类。最著名的亚类是含有苯并 -γ- 吡咯酮骨架的化合物。黄酮类化合物通常以糖苷的形式出现，它们扮演苷元的角色，通过糖苷键与碳水化合物基团相连[15-17]。类黄酮分子中酚基的存在使它们具有更强的抗自由基活性，因为在清除过程中形成的自由基是共振稳定的。在蜂产品中发现的黄酮类化合物及其糖苷的例子见图1。
Figure 1 图1The examples of flavonoids and their glycosides detected in bee products. Quercetin, naringenin, isorhamnetin, and kaempferol: detected in propolis, bee pollen, royal jelly; galangin and pinocembrin: detected in propolis and bee pollen; pinobanksin: detected in propolis and royal jelly; luteolin, apigenin, and rutin: detected in propolis, bee pollen, and royal jelly; catechin and delphinidin: detected in bee pollen; daidzein: detected in propolis [ 蜂产品中黄酮类化合物及其苷类成分的检测。槲皮素、柚皮素、异鼠李素和山奈素: 在蜂胶、蜂花粉、蜂王浆、高良姜素和松节苷中检测出: 在蜂胶和蜂花粉中检测出: 在蜂胶和蜂王浆中检测出: 在蜂胶、蜂花粉和蜂王浆中检测出: 在蜂胶、蜂花粉和蜂王浆中检测出: 在儿茶素和飞燕草素中检测出: 在蜂花粉中检测出: 在蜂胶中检测出: 在蜂胶中检测出:29, 32, 74, 78, 83, 103].
The phenolic acids are compounds possessing carboxylic and phenol groups. Recently, a growing concern in their possible application for human health protection has been observed, considering their antioxidant activities including the prevention of oxidation processes and generation of oxygen species as well as chelating prooxidative metals . The examples of phenolic acids and their derivatives found in bee products are presented in Figure 2.
(b)Figure 2 图2The examples of phenolic acids and their derivatives found in bee products: (a) benzoic acid derivatives and (b) cinnamic acid derivatives. Protocatechuic acid, syringic acid, gallic acid, 在蜜蜂产品中发现的酚酸及其衍生物的例子: (a)苯甲酸衍生物和(b)肉桂酸衍生物。原儿茶酸，丁香酸，没食子酸,p-coumaric acid: detected in propolis and bee pollen; caffeic acid and ferulic acid: detected in propolis, bee pollen, and royal jelly; artepillin C, chlorogenic acid, and 3,5-dicaffeoylquinic acid: detected in propolis [ – 香豆酸: 在蜂胶和蜂花粉中检测到; 咖啡酸和阿魏酸: 在蜂胶、蜂花粉和蜂王浆中检测到; 在蜂胶中检测到青蒿素 c、绿原酸和3,5- 二咖啡酰奎宁酸28, 41, 71, 74, 78, 102, 139].
To compounds without phenolic character being responsible for the antioxidant capacity of propolis (especially Brazilian one) belong amyrins [19, 20]. α– and β-amyrins belong to triterpenoids of plant origin. They have been reported to exhibit numerous beneficial properties including antiapoptotic, antioxidant, anti-inflammatory, and antifibrotic as well as gastro- and hepatoprotective effects. Studies have revealed the possible application of β-amyrin in Parkinson’s disease therapy [21, 22]. The examples of amyrins found in bee propolis are presented in Figure 3.
对于没有酚性质的化合物负责蜂胶(特别是巴西的)的抗氧化能力属于阿米林[19,20]。α-和 β- 淀粉蛋白属于植物起源的三萜类化合物。据报道，它们具有许多有益的特性，包括抗凋亡、抗氧化、抗炎症、抗纤维化以及胃和肝保护作用。研究揭示了 β- 淀粉蛋白在帕金森病治疗中的可能应用[21,22]。蜂胶中发现的淀粉样蛋白的例子如图3所示。
Considering the antioxidant activity of royal jelly hydroxy dicarboxylic fatty acids with 8–12 carbon atoms in the chain and their derivatives is worth mentioning. The major fatty acid is 10-hydroxydecanoic acid (10-HDA) whose presence has not been reported in any other natural raw material or even in any other product of apiculture . Other carboxylic acids included in RJ are 10-hydroxy-2-decenoic acid (10H2DA) and sebacic acid (SA)  (Figure 4).
考虑到含8-12个碳原子的蜂王浆羟基二羧酸及其衍生物的抗氧化活性，值得一提。主要的脂肪酸是10- 羟基癸酸(10-HDA) ，其存在尚未报告在任何其他天然原料，甚至在任何其他养蜂产品中。其他包括在 RJ 的羧酸是10- 羟基 -2- 癸烯酸(10H2DA)和癸二酸(SA)(图4)。
In scientific research, propolis and bee pollen extracts are used instead of the raw substance due to the fact that they contain higher amounts of bioactive components . However, the application of solvents of different polarities affects the composition of the obtained extracts as the components of bee products possess diverse structures, and while hydrophilic ones are better soluble in polar solvents like alcohols, those with hydrophobic properties exhibit greater affinity to nonpolar solvents like hydrocarbons. The properties of the extract depend strongly not only on the solvent used but also on extraction conditions, that is, time and temperature as well [13, 25].
Having regarded the importance of acquiring drugs from natural sources, this review is aimed at updating the current state of the knowledge of antioxidant capacity of the selected bee products, namely, propolis, bee pollen, and royal jelly, and of their possible medical applications as natural substances capable of counteracting the effects of oxidative stress underlying pathogenesis of numerous diseases, such as neurodegenerative disorders, cardiovascular diseases, diabetes, and cancer, as well as negative effects of different harmful factors and drugs. Moreover, particular attention has been attributed to the understanding of the mechanisms underlying possible antioxidant properties of bee products. The influence of bee species, plant origin, geographic location, and seasonality, as well as type of extraction solutions on the composition of bee products extracts were all discussed.
2. Propolis: “Bee Glue”
2. 蜂胶: “蜂胶”
Propolis, generally known as the “bee glue,” is a resinous mixture that honey bees produce by mixing their saliva containing certain enzymes and beeswax with exudate gathered mainly from leaf and flower buds, stems, and bark cracks of numerous species of trees. The word propolis is derived from two Greek words pro and polis, which mean “defense” and “city” or “community,” respectively. Bees use it mainly as a sealant and a disinfecting material. Propolis is used for sealing holes and cracks, smoothing the inner surface, and retaining internal temperature of the beehive as well as for preventing weathering (e.g., it is used for decreasing the size of the outlet opening during periods of cold weather) and invasion of predators [3, 26]. Due to its antimicrobial activity, it also contributes to an aseptic internal environment and is used to cover (“mummify”— to prevent decay) the body of dead pests that have invaded the hives (e.g., shrews and mice), which are too big to be removed outside .
蜂胶，通常被称为“蜂胶” ，是一种树脂混合物，蜂蜜产生的唾液含有某些酶和蜂蜡与渗出物收集主要从叶子和花蕾，茎，和树皮裂缝的许多种树木。蜂胶这个词来源于两个希腊词 pro 和 polis，分别意为“防御”和“城市”或“社区”。蜜蜂主要用它作为密封剂和消毒材料。蜂胶用于密封孔洞和裂缝，磨光内表面，保持蜂巢的内部温度，以及防止风化(例如，在寒冷天气期间用于缩小出口的大小)和天敌入侵[3,26]。由于其抗菌活性，它也有助于无菌的内部环境，并用于覆盖(“木乃伊化”ーー防止腐烂)侵入蜂巢的死害虫尸体(例如，鼩鼱和老鼠) ，这些害虫体型太大，无法移出蜂巢。
2.1. Composition of Propolis
Raw propolis is typically composed of 50–60% of resins and balms (including phenolic compounds), 30–40% of waxes and fatty acids, 5–10% of essential oils, 5% of pollen, and about 5% of other substances including amino acids, micronutrients, and vitamins (thiamin, riboflavin, pyridoxine, C, and E) [20, 27]. According to the literature data, more than 300 compounds belonging to polyphenols, terpenoids, steroids, sugars, amino acids, and others have been identified in propolis [3, 28].
原始蜂胶通常由50-60% 的树脂和香料(包括酚类化合物) ，30-40% 的蜡和脂肪酸，5-10% 的精油，5% 的花粉，以及大约5% 的其他物质，包括氨基酸，微量营养素，和维生素(硫胺素，核黄素，吡哆醇，c，和 e)[20,27]组成。根据文献资料，在蜂胶中已经鉴定出300多种化合物，包括多酚、萜类、甾体、糖类、氨基酸等。
Propolis from the temperate zone all over the world (Europe, nontropic regions of Asia, North America, and continental Australia) is classified as the poplar type propolis since it originates mainly from the bud exudates of Populus spp., most often P. nigra L. The main biologically active components of this type of propolis are flavonoids (flavones and flavanones), phenolic acids (cinnamic acid), and their esters [27–29]. Birch propolis, found in Russia, originates from Betula verrucosa Ehrh. and similarly contains flavones and flavonols (but not the same as in poplar propolis) . Mediterranean propolis is characteristic of subtropic regions like Greece, Greek islands, Sicily, Malta, Cyprus, Croatia, and Algeria. It originates mainly from the resin of Cupressus sempervirens(commonly known as the Mediterranean or Italian cypress) and is characterized by relatively high amounts of diterpenes [28, 29]. In tropical zones, in turn, several types of propolis derived from many different sources have been identified. For example, in Brazil, there are 13 different types of propolis including green, red, and brown ones, whose main sources are Baccharis dracunculifolia, Dalbergia ecastaphyllum , and Hyptis divaricata, respectively. The most popular is the green one, which owes its color to the chlorophyll occurring in young tissues and nonexpanded leaves of B. dracunculifolia and collected by the bees . This type of propolis is rich in derivatives of phenylpropanoids (e.g., artepillin C) and diterpenes, whereas flavonoids occur in small amounts . The red propolis is characterized by the presence of numerous flavonoids (formononetin, liquiritigenin, pinobanksin-3-acetate, pinobanksin, luteolin, rutin, quercetin, pinocembrin, daidzein, and isoliquiritigenin), which are found in the resinous exudates from the surface of D. ecastaphyllum [30, 32]. This type of propolis is also characteristic of Cuba and Mexico . The brown propolis is mainly produced in northeastern Brazil from H. divaricata . Other examples of tropical propolis include the one originating from resin exuded by the flowers of Clusia sp. found in Cuba and Venezuela—with its main constituents being derivatives of benzophenones—and “Pacific” propolis originating from the tropical tree Macaranga tanarius found on Pacific Ocean tropical islands (Taiwan, Okinawa, and Indonesia), which chemical makers are C-prenylflavanones [29, 33].
产自世界各地温带(欧洲、亚洲、北美和澳大利亚大陆等非热带地区)的蜂胶主要来源于黑杨属植物的芽分泌物，因此被归为杨树型蜂胶。这类蜂胶的主要生物活性成分是黄酮类(黄酮类和黄酮类)、酚酸类(肉桂酸)及其酯类(27-29)。桦木蜂胶产于俄罗斯，原产于 Betula。类似地，它还含有黄酮和黄酮醇(但不同于杨树蜂胶)。地中海蜂胶的特点是亚热带地区，如希腊，希腊群岛，西西里岛，马耳他，塞浦路斯，克罗地亚和阿尔及利亚。它主要来源于地中海柏木的树脂(通常称为地中海或意大利柏树) ，拥有属性中二萜的含量相对较高[28,29]。在热带地区，从许多不同来源的蜂胶已经被鉴定出几种类型。例如，在巴西，有13种不同类型的蜂胶，包括绿色、红色和棕色的蜂胶，它们的主要来源分别是龙纹芽孢杆菌(Baccharis dracunculifolia)、黄檀(Dalbergia ecastaphylum)和神木(Hyptis divaricata)。最受欢迎的是绿色的，它的颜色来自于存在于龙血竭幼嫩组织和未展开叶片中的叶绿素，这些叶片是由蜜蜂采集的。这种蜂胶含有丰富的苯丙素类衍生物(如青蒿素 c)和二萜类化合物，而黄酮类化合物含量很少。红色蜂胶中含有大量的黄酮类化合物(拥有属性，甘草素，3- 乙酸松黄素，木犀草素，芦丁，槲皮素，松黄素，大豆苷元和异黄酮素) ，这些黄酮类化合物存在于东北大戟表面的树脂性渗出物中。这种类型的蜂胶也是古巴和墨西哥的特色。棕色蜂胶主要产于巴西东北部，原产于印度。热带蜂胶的其他例子包括一种原产于古巴和委内瑞拉的克鲁西亚花中渗出的树脂，其主要成分是二苯甲酮的衍生物，以及一种原产于太平洋热带岛屿(台湾、冲绳和印度尼西亚)的热带血桐的“太平洋”蜂胶，其化学制造商是 C-prenylflavanones [29,33]。
2.2. Relationships between the Antioxidant Capacity of Propolis and Its Composition
Antioxidant properties of propolis have been fully investigated and proven with the use of DPPH, ABTS+, FRAP, and ORAC methods [3, 19, 20, 29, 31, 34–38]. In the same in vitro studies, the antioxidant capacity of propolis extracts was found to be similar to that of the synthetic antioxidant butylated hydroxytoluene or ascorbic acid [4, 20]. Importantly, the antioxidant capacity of propolis is dependent on its content, but the studies aiming at finding the distinct relationships between these two parameters are not consistent [3, 35, 36]. In general, according to the literature data, the total phenolic content of propolis extracts ranged from about 30 to 200 mg of gallic acid equivalents (GAE)/g of dry weight, and the flavonoid content ranged from about 30 to 70 mg of quercetin equivalents (QE)/g, whereas DPPH free radical-scavenging activity ranged from about 20 to 190 μg/mL [3, 19, 20, 31, 35, 36, 38]. The phenolic compounds, but the different ones than the flavonoids, are believed to be responsible for the antioxidant activity of Brazilian propolis. According to Zhang et al. , 3,4,5-tricaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and artepillin C seem to be responsible for the strong antioxidant activity of Brazilian green propolis. Unlike in the case of Brazilian propolis, the antioxidant activity of poplar propolis appears to be largely influenced by both total polyphenol and total flavonoid contents [3, 34, 38]. The results obtained by Fabris et al.  indicated that European (Italy and Russia) propolis samples had similar polyphenolic composition and consequently similar antioxidant activity, while Brazilian propolis possessed lower polyphenolic amount and thus antioxidant properties. In general, there seems to be a big problem with the standardization of propolis composition—this results from the fact that this is highly dependent on many factors, such as bee species, plant origin, geographic location, temperature variation, and seasonality, as well as storage conditions [3, 19, 20, 27, 31, 36, 40]. Recently, for example, Bonamigo et al. [19, 20] have studied the antioxidant activity of the ethanol extract of Brazilian propolis samples (collected from the same region) depending on bees’ species, Scaptotrigona depilis, Melipona quadrifasciata anthidioides, Plebeiadroryana, and Apis mellifera. The studied samples were shown to differ in composition as well as in both free radical-scavenging activity and ability to inhibit lipid peroxidation. In general, propolis obtained from A. mellifera revealed the highest activity. Calegari et al. , in turn, found that Brazilian propolis samples produced in both March and April showed a difference in color and higher content of total phenolic compounds as well as antioxidant capacity than those produced in May and June, which indicated that the chemical composition of propolis depended on the month of production—this effect can be explained by variations in temperature. The researchers also reported that bees’ colonies, which received food supplementation every three days throughout the year, displayed significantly higher total phenolic and flavonoids content as well as antioxidant capacity than those bereaved of this supplementation .
利用 DPPH 法、 ABTS + 法、 FRAP 法和 ORAC 法[3,19,20,29,31,34-38]对蜂胶的抗氧化性能进行了充分的研究和验证。在同样的体外研究中，发现蜂胶提取物的抗氧化能力与合成抗氧化剂2,6-二叔丁基对甲酚或抗坏血酸相似[4,20]。重要的是，蜂胶的抗氧化能力取决于其含量，但是旨在发现这两个参数之间的不同关系的研究并不一致[3,35,36]。一般来说，根据文献资料，蜂胶提取物的总酚含量约为30ー200mg 没食子酸当量(GAE)/g 干重，黄酮含量约为30ー70mg 槲皮素当量(QE)/g，而 DPPH 自由基清除活性约为20ー190μg/mL [3,19,20,31,35,36,38]。酚类化合物，但不同于黄酮类化合物，被认为是负责巴西蜂胶的抗氧化活性。根据 Zhang 等人的研究 ，3,4,5- 三咖啡酰奎宁酸，3,5- 二咖啡酰奎宁酸，4,5- 二咖啡酰奎宁酸，和青蒿素 c 似乎是巴西绿色蜂胶强大的抗氧化活性的原因。与巴西蜂胶不同，杨木蜂胶的抗氧化活性似乎主要受总多酚和总黄酮含量的影响[3,34,38]。Fabris 等人的结果表明，欧洲(意大利和俄罗斯)蜂胶样品具有相似的多酚组成，因此具有相似的抗氧化活性，而巴西蜂胶的多酚含量较低，因此具有抗氧化性能。一般来说，蜂胶成分的标准化似乎存在一个大问题ーー这是因为蜂胶的标准化高度依赖于许多因素，如蜜蜂种类、植物起源、地理位置、温度变化和季节性，以及贮藏条件[3,19,20,27,31,36,40]。最近，例如，Bonamigo 等人[19,20]研究了巴西蜂胶样品(采自同一地区)乙醇提取物的抗氧化活性，这取决于蜜蜂的种类，toscaptrigona depilis，Melipona quadriata idioides，Plebeiadroryana 和意大利蜜蜂。研究表明，样品的组成不同，以及在自由基清除活性和抑制脂质过氧化的能力。一般来说，蜂胶的活性最高。与5月和6月相比，3月和4月生产的巴西蜂胶样品在颜色和总酚类化合物含量以及抗氧化能力方面都有所不同，这表明蜂胶的化学成份取决于生产的月份—- 这种影响可以用温度的变化来解释。研究人员还报告说，一年中每三天接受一次食物补充的蜜蜂蜂群，其总酚类和类黄酮含量以及抗氧化能力显著高于那些失去这种补充的蜜蜂。
Moreover, both the chemical composition and biological properties of propolis extracts are highly dependent on the type of solvents used for the extraction [3, 31, 34]. The most commonly used solvent for the extraction of propolis is aqueous ethanol (particularly at concentration of 70–75%), followed by others, such as ethyl ether, water, methanol, hexane, and chloroform. Sun et al.  showed that extraction yields (the ratio of the weight of the dry extract to the weight of the raw extract) of Beijing propolis ranged from 1.8% to 51% and exhibited the tendency to increase along with the enhancement of the ethanol concentration. The total polyphenol and total flavonoid content distinctly varied and ranged from 6.68 to 164.20 mg GAE/g and 4.07 to 282.83 mg of rutin equivalents (RE)/g, respectively, and the highest concentration was observed in 75% ethanol solvents, a little bit lower in 95% and 100% ethanol solvents and the lowest one in water solution. The 75% extract also demonstrated the highest antioxidant capacity measured by DPPH, ABTS, FRAP, oxygen radical absorbance capacity (ORAC), and cell antioxidant activity (CAA) methods. In general, polar solvents allow obtaining better antioxidant properties than the nonpoplar ones. However, considerable differences were observed even in the case of the application of solvents of similar polarity or the same solvent for the extraction of different types of propolis samples , pointing to the possible influence of other parameters as well as the impact of the molecular structure of solvents. Bittencourt et al. , for example, showed that that partition with dichloromethane enhanced the extraction of antioxidant compounds, especially in brown propolis, whereas partition with hexane significantly decreased their amount in green propolis extract (Table 1).
此外，蜂胶提取物的化学成份和生物特性都高度依赖于提取溶剂的类型[3,31,34]。蜂胶最常用的提取溶剂是乙醇水溶液(特别是70-75% 的浓度) ，其次是乙醚、水、甲醇、正己烷和氯仿。结果表明，北京蜂胶的提取率(干浸膏重量与粗浸膏重量的比值)在1.8% ー51% 之间，且随着乙醇浓度的增加呈上升趋势。总多酚和总黄酮含量变化很大，分别为6.68ー164.20 mg GAE/g 和4.07ー282.83 mg/g，在75% 乙醇溶剂中含量最高，在95% 和100% 乙醇溶剂中略低，在水溶液中含量最低。用 DPPH 法、 ABTS 法、 FRAP 法、氧自由基吸收能力(ORAC)法和细胞抗氧化能力(CAA)法测定，75% 提取物的抗氧化能力最高。一般来说，极性溶剂比非杨木溶剂具有更好的抗氧化性能。然而，即使在使用相似极性的溶剂或使用相同溶剂提取不同类型的蜂胶样品的情况下，也观察到了相当大的差异，指出了其他参数可能的影响以及溶剂分子结构的影响。例如，Bittencourt et al. 表明，与二氯甲烷的分配增强了抗氧化剂化合物的提取，尤其是在棕色蜂胶中，而与正己烷的分配则显著降低了绿色蜂胶提取物中抗氧化剂化合物的含量(表1)。
|Source资料来源Type of bee products 蜂产品种类The used solvents 用过的溶剂The dependence between the used extraction solvent and the properties of the obtained extract 所用萃取溶剂与所得萃取物性质之间的相关性Bittencourt et al. [等人Green and brown Brazilian propolis 绿色和棕色的巴西蜂胶Ethanol (95%) extraction, evaporation, and dissolving in 80% ethanol and then partitioning with hexane or dichloromethane. 乙醇(95%)萃取、蒸发、溶解于80% 乙醇中，然后用正己烷或二氯甲烷分配Antioxidant activity showed considerable differences depending on the used solvent and propolis type. 抗氧化活性表现出相当大的差异，取决于使用的溶剂和蜂胶类型|
In DPPH assay, the strongest antioxidant activity was found in dichloromethane and ethanol extracts of green propolis as well as dichloromethane extract of brown propolis with the IC50 values at least two times less than in the other cases, namely, in hexane extract of brown propolis, ethanol extract of brown propolis, and hexane extract of green propolis. 在 DPPH 法测定中，最强抗氧化活性的是绿蜂胶的二氯甲烷和乙醇提取物，以及棕蜂胶的二氯甲烷提取物，其 IC50值至少比棕蜂胶的正己烷提取物、棕蜂胶的乙醇提取物和绿蜂胶的正己烷提取物低2倍Narimane et al. [等人Algerian propolis collected from Beni Belaid, Jijel (northeast of Algeria) 采自吉杰勒贝尼贝莱德的阿尔及利亚蜂胶CH 甲烷2Cl 女名女子名2-MeOH (1 : 1, – MeOH (1:1,v/v) extraction followed by MeOH-H )萃取，然后进行 MeOH-H2O (70 : 30, o (70:30,v/v) one, concentrating and dissolving in boiling water. Then the water solution underwent successive extraction by using of chloroform, ethyl acetate, and )1，浓缩后放入沸水中溶解。然后用氯仿、乙酸乙酯和乙酸乙酯连续萃取水溶液n-butanol. 丁醇The DPPH, ABTS, FRAP, and CUPRAC methods were applied to determine antioxidant activity. 采用 DPPH 法、 ABTS 法、 FRAP 法和 CUPRAC 法测定抗氧化活性
The ethyl acetate and n-butanol extracts proved to be the most effective ones. 乙酸乙酯和正丁醇提取物是最有效的提取物Sun et al. [孙等人Beijing propolis 北京蜂胶科技有限公司Water, 25%, 50%, 75%, 95%, and 100% ethanol. 水、25% 、50% 、75% 、95% 和100% 乙醇The 75% extract demonstrated the highest antioxidant capacity measured by DPPH, ABTS, FRAP, ORAC, and CAA methods. 用 DPPH 法、 ABTS 法、 FRAP 法、 ORAC 法和 CAA 法测定75% 提取物的抗氧化能力最高LeBlanc et al. [ 勒布朗等人Six pollen types (mesquite, yucca, palm, terpentine bush, mimosa, and chenopod) collected in Arizona between March and November 在3月至11月期间，在亚利桑那州采集了6种花粉类型(豆科、丝兰属、棕榈属、松节油灌木属、含羞草属和藜科)Water, methanol, ethanol, propanol, 2-propanol, acetone, dimethylformamide, and acetonitrile. 水、甲醇、乙醇、丙醇、异丙醇、丙酮、丙酮和乙腈Antioxidant activity showed considerable differences depending on the used solvent. 抗氧化活性表现出相当大的差异取决于使用的溶剂
In FRAP assay, methanol and dimethylformamide extracts displayed the greatest activity values, whereas those obtained with the application of acetonitrile displayed the lowest ones. Acetone extracts were also found to possess low activity in the case of most types of bee pollen. 在 FRAP 分析中，甲醇和丙酮提取物的活性最高，而乙腈提取物的活性最低。丙酮提取物也被发现具有低活性的情况下，大多数类型的蜂花粉
Similar results were obtained with using DPPH assay. DPPH 法也得到了类似的结果Silva et al. [ Silva 等Pollen loads collected from 花粉载量采集自Melipona rufiventris 梅利波娜 · 卢菲文特里斯 (stingless bees) colonies in Brazil (无刺蜜蜂)在巴西的殖民地Fractional extraction with subsequent using of ethanol, n-hexane, and ethyl acetate. 用乙醇、正己烷和乙酸乙酯进行分步萃取The DPPH method was applied to determine antioxidant activity. 采用 DPPH 法测定抗氧化活性
The ethyl acetate extract proved to be the most effective one. 乙酸乙酯提取物是最有效的提取物
The ethanol extract showed much less (more than six times) efficiency, whereas the capacity of the hexane extract was slight (practically inactive). 乙醇提取物的效率要低得多(超过六倍) ，而己烷提取物的容量很小(实际上不活跃)Chantarudee et al. [ 尚塔鲁迪等Bee pollen collected in Thailand in the summer (June), its main component being identified as pollen of corn ( 泰国夏季(月日)采集的蜂花粉，其主要成分为玉米花粉(Z. mays L.)Subsequent application of 80% methanol, dichloromethane, and hexane. 80% 甲醇、二氯甲烷和正己烷的后续应用The antioxidant activity of the obtained extracts was estimated by DPPH assay. 用 DPPH 法测定提取物的抗氧化活性
The hexane extract proved to be completely inactive, whereas that obtained with using dichloromethane showed the best antioxidant properties, comparable even with the reference antioxidant—ascorbic acid. 正己烷提取物完全失活，而二氯甲烷提取物的抗氧化性能最好，甚至可与参考抗坏血酸相媲美Maruyama et al. [ Maruyama 等Bee pollen from 蜜蜂花粉Cistus 女名女子名 sp. of Spanish origin. Bee pollen from起源于西班牙的蜂花粉Brassica 芸苔属 sp. of Chinese origin 起源于中国的Water and 95% ethanol 水和95% 的乙醇In rats with induced hind paw edema, the oral administration of bee pollen water extract had practically no effect, while ethanol extract displayed the greatest effectiveness in the inhibition of paw edema. 在大鼠足肿胀模型中，蜂花粉水提取物的口服给药几乎没有作用，而乙醇提取物在抑制足肿胀方面效果最好
Table 1 表一The results of the research on the dependence between the solvent used for the extraction and the antioxidant properties of the obtained propolis and bee pollen extracts. 研究了蜂胶和蜂花粉提取物的抗氧化性能与提取溶剂的相关性
Importantly, despite numerous differences in its composition, propolis extract always possesses antioxidant properties. Even the aqueous extracts of propolis were shown to display antioxidant capacity in cell culture and animal studies [9, 12].
2.3. Antioxidant Effect of Propolis in Human Studies
Most of the studies regarding antioxidant properties of propolis have been performed on cell culture or animals. In the available literature, there are only a few studies investigating the antioxidant effect of propolis in humans.
Mujica et al.  evaluated the effects of the oral administration (twice daily, 15 drops each time, 90 days) of commercially available propolis solution (Beepolis®) on the oxidative status and lipid profile in a human population in Chile. The 90-day propolis supplementation resulted in a 67% decrease in the amount of thiobarbituric acid reactive substances (TBARS; lipid peroxidation derivative products) and 175% increase in reduced glutathione (GSH) level compared to the baseline. Net changes of both studied parameters were significantly higher in propolis supplemented group than those observed in the placebo group. Moreover, an increase in the HDL concentration on the 90th day of propolis supplementation compared to the baseline value was observed. The authors concluded that propolis supplementation appeared to have positive effects on oxidative status and the improvement of HDL and may thus reduce the risk of cardiovascular events.
Mujica et al. 评估了口服给药(每天两次，每次15滴，90天)的蜂胶溶液(蜂胶)对智利人体的氧化状态和脂质状况的影响。补充90天蜂胶导致硫代巴比妥酸活性物质(TBARS; 脂质过氧化衍生物产品)的数量减少了67% ，与基线相比谷胱甘肽(GSH)水平增加了175% 。蜂胶补充组两项参数的净变化明显高于安慰剂组。此外，在补充蜂胶的第90天，高密度脂蛋白浓度比基线值有所增加。作者得出结论，蜂胶补充剂对氧化状态和提高高密度脂蛋白有积极作用，因此可能降低心血管事件的风险。
Jasprica et al. , in turn, investigated the issue of the possible influence of 30-day supplementation with commercially available powdered propolis extract (a total daily dose of flavonoids was 48.75 mg) on antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and a lipid peroxidation marker—malondialdehyde (MDA)—in healthy individuals. In the male group, after 15 days of propolis treatment, a 23.2% decrease in MDA level was observed, whereas after 30 days, a 20.9% increase in SOD activity was found. Interestingly, MDA concentration in the end of treatment was similar to the baseline value. The propolis treatment had no effect on any of the studied parameters in women (). The authors concluded that the effect of propolis was both time and gender dependent and suggested a possibility of existence of only the transitory effect of propolis ingestion on lipid peroxidation.
另一方面，研究了30天补充市售蜂胶粉提取物(每日总黄酮量为48.75 mg)对健康人体内抗氧化酶如超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)、过氧化氢酶(CAT)和脂质过氧化丙二醛(MDA)的影响。在雄性组中，蜂胶处理15天后，MDA 含量下降23.2% ，而处理30天后 SOD 活性上升20.9% 。有趣的是，治疗结束时 MDA 的浓度与基线值相似。蜂胶治疗对女性的任何研究参数都没有影响()。作者得出结论，蜂胶的影响是时间和性别相关的，并提出了一种可能性，即只存在摄入蜂胶对脂质过氧化的短暂影响。
The effect of Brazilian green propolis supplementation on antioxidant status in patients with type 2 diabetes mellitus (T2DM) was studied by Zhao et al. . The propolis administration (900 mg/day, 18 weeks) was associated with an increase in serum levels of GSH and total polyphenols and reduction in serum carbonyls (protein oxidation markers) as well as lactate dehydrogenase activity. Moreover, the Brazilian green propolis group revealed a decreased TNF-αserum level and significantly increased IL-1β and IL-6 sera levels. However, the propolis treatment did not affect serum glucose, glycosylated hemoglobin, insulin, aldose reductase, or adiponectin. The above results indicate that propolis affects the oxidative stress in type 2 diabetic patients but not the parameters of diabetes.
赵等人研究了补充巴西绿蜂胶对2型糖尿病(T2DM)患者抗氧化状态的影响。蜂胶剂量(900毫克/天，18周)与血清谷胱甘肽和总多酚水平升高、血清羰基化物(蛋白质氧化标志物)减少以及乳酸脱氢酶活性升高有关。此外，巴西绿蜂胶组血清 tnf-α 水平降低，血清 il-1β 和 IL-6水平显著升高。然而，蜂胶治疗不影响血糖，糖化血红蛋白，胰岛素，醛糖还原酶，或脂联素。上述结果表明，蜂胶对2型糖尿病患者的氧化应激有影响，但对糖尿病参数无影响。
2.4. Neuroprotective Effects of Propolis
Since mitochondrial damage and oxidative stress are critical events in neurodegeneration, in recent years, it has been suggested that antioxidant properties of the constituents of propolis may contribute to its neuroprotective effects. The effect of water-extracted brown propolis (WEBPs), from two regions of Iran, against cerebral ischemia-induced oxidative injury in a mouse model of stroke was studied by Bazmandegan et al. . Regardless of the region of origin or the used doses, WEBP treatment resulted in a significant restoration of antioxidant enzymes activity and a decrease in both lipid peroxidation and the infarct volume, compared to the control group. Moreover, the treatment was associated with an improvement of neurological deficits measured with the Bederson scale and sensorimotor function measured with sticky removal tape test (Table 2). In another study performed on SH-SY5Y cells , it was found that the pretreatment with Brazilian green propolis reduced the H2O2-induced mitochondria-derived intracellular reactive oxygen species (ROS) generation as well as 8-oxo-2′-deoxyguanosine (8-oxo-dG, the DNA oxidative damage marker) immunofluorescence signal intensity. Propolis was also shown to increase the expression of the critical factors of synapse efficacy, brain-derived neurotrophic factor (BDNF), and activity-regulated cytoskeleton-associated protein (Arc). The obtained outcomes allowed the authors to suggest that propolis displays protective abilities against neurodegenerative damage, related to cognitive impairment caused by Alzheimer’s disease or aging, via its antioxidant action (Table 2). This seems to be consistent with the results obtained by Nanaware et al. , who studied the neuroprotective activity of the macerated ethanolic extract of Indian propolis (MEEP) in rat model of Alzheimer’s disease. MEEP significantly reversed the cognitive impairment of β amyloid-induced rats, which, among other things, was associated with increased antioxidant and decreased MDA levels. In addition, MEEP administration resulted in dose-dependent acetylcholinesterase inhibition, increased brain monoamine level, and improved memory deficits (assessed by increased BDNF level), which suggested that multiple mechanisms might be involved in that neuroprotective effect of propolis (Table 2).
由于线粒体损伤和氧化应激是神经退行性疾病的关键事件，近年来，有人认为蜂胶成分的抗氧化特性可能有助于其神经保护作用。用巴兹曼德甘等研究了伊朗两个地区的水提棕色蜂胶对脑缺血诱导的氧化损伤的影响。与对照组相比，不管起源于哪个地区或使用的剂量，WEBP 治疗导致了抗氧化酶活性的显著恢复和脂质过氧化和梗死体积的减少。此外，治疗与改善神经功能缺陷测量的 Bederson 量表和感觉运动功能测量的粘性拆除胶带测试(表2)。另一项对 SH-SY5Y 细胞的研究发现，巴西绿蜂胶预处理可以降低 h2o2诱导的线粒体衍生的细胞内活性氧类(ROS)的产生，以及8- 氧代脱氧鸟苷(8-oxo-2′-deoxyguanosine，DNA 氧化损伤标志物)的免疫荧光信号强度。蜂胶也被证明可以增加突触功效的关键因子的表达，脑源性神经营养因子和活性调节的细胞骨架相关蛋白(Arc)。研究结果表明，蜂胶具有抗氧化作用，可以防止神经退行性损伤，这种损伤与阿尔茨海默病或衰老引起的认知障碍有关(表2)。这似乎与 Nanaware 等人在阿尔茨海默病大鼠模型中研究印度蜂胶乙醇提取物(MEEP)神经保护活性的结果一致。MEEP 显著逆转了 β 淀粉样蛋白诱导的大鼠的认知障碍，除其他外，这与抗氧化剂增加和 MDA 水平降低有关。此外，给予 MEEP 可导致剂量依赖性乙酰胆碱酯酶抑制，增加脑单胺水平，改善记忆缺陷(通过增加脑源性神经营养因子水平来评估) ，这表明蜂胶的神经保护作用可能涉及多种机制(表2)。
|Source 资料来源Toxic or harmful factor 中毒或有害因素Harmful effects of an applied factor 应用因素的有害影响The type of propolis and the way of application 蜂胶的种类及应用方法Effects of propolis coadministration 蜂胶辅助给药效果观察Neuroprotective effect of propolis 蜂胶的神经保护作用 Bazmandegan et al. [ 巴兹曼德甘等人Cerebral ischemia-induced oxidative injury in a mouse model of stroke 脑缺血致脑卒中小鼠氧化损伤的实验研究↑ MDA 温度|
↑ SOD and SOD/GPx ratio SOD ↑与 SOD/gpx 比值
↓ GPx in brain 4. 大脑的↓ GPxWater-extracted Iran brown propolis; from two regions of Iran; 100 and 200 mg/kg; 水提取的伊朗棕色蜂胶; 来自伊朗两个地区; 100和200毫克/公斤;i.p. at 48, 24, and 1 h before and 4 h after the induction of ischemia 分别在诱导缺血前48,24,1h 和4h↓ MDA 4. ↓ MDA
↓ SOD and SOD/GPx ratio ↓ SOD 和 SOD/gpx 比率
↑ GPx 图像∗∗ 第 x 次/次 in brain在大脑里Ni et al. [等人H2O2-induced neurotoxicity, human neuroblastoma SH-SY5Y cells (100 目的: 研究人神经母细胞瘤 SH-SY5Y 细胞(100μM for 4 h or 1 h for ROS) M 为4小时，ROS 为1小时)↑ ROS in mitochondria 线粒体中↑活性氧
↑ 8-oxo-dG, the DNA oxidative damage marker DNA 氧化损伤标记物↑8-oxo-dG
↓ Cell viability 4. ↓细胞活力Methanol extract of Brazilian green propolis, 10 巴西绿蜂胶的甲醇提取物，10μg/mL pretreatment for 2 h (or 1 h for ROS) g/mL 预处理2小时(ROS 预处理1小时)↓ ROS in mitochondria 4. 线粒体中的 ROS
↓ 8-oxo-dG ↓8-oxo-dG
↑ Cell viability ↑细胞活力Nanaware et al. [ Nanaware 等β-Amyloid 25–35-induced Alzheimer’s disease model in rats, (10 – 淀粉样蛋白25-35诱导的大鼠阿尔茨海默病模型，(10μg/rat injected bilaterally) G/rat 双侧注射)↓ SOD, GSH, CAT, NO 4. 名片↓ SOD，GSH，CAT，NO
↑ MDA in brain 脑内 MDA ↑Macerated ethanolic extract of Indian propolis; 100, 200, and 300 mg/kg b.w., 印度蜂胶浸渍乙醇提取物; 100,200和300毫克/千克,p.o. (posttreatment after 14 days); 21 days . (治疗后14天) ; 21天↑ SOD, GSH, CAT, NO SOD，GSH，CAT，NO
↓ MDA in brain 4. 脑海中的 MDA
All doses were effective; the effect slightly increased with increasing dose 所有剂量均有效，效果随剂量增加略有增加Jin et al. [金等人6-Hydroxydopamine-induced oxidative stress in human neuroblastoma SH-SY5Y cells (50 6- 羟基多巴胺诱导人神经母细胞瘤 SH-SY5Y 细胞产生氧化应激μM for 24 h) M 代表24小时)↑ ROS 活性氧
↑ MDA 温度
↓ SOD 名片↓ SOD
↓ Bcl-2/Bax ratio ↓ Bcl-2/Bax 比例Pinocembrin; 1, 5, and 25 Pinocembrin; 1岁，5岁，25岁μM pretreatment for 4 hours M 预处理4小时↓ ROS∗∗ 第 x 次/次
↓ MDA 4. ↓ MDA∗∗ 第 x 次/次
↑ SOD∗∗ 第 x 次/次
↑ Bcl-2/Bax ratio Bcl-↑-2/Bax 比值∗∗ 第 x 次/次
↑ Nrf2 translocation Nrf2易位∗∗ 第 x 次/次
↑ HO-1 and 转座子 HO-1和γ-GCS expression – GCS 表达∗∗ 第 x 次/次de Oliveira et al. [ 奥利维拉等人Paraquat-induced neurotoxicity in SH-SY5Y cells (100 百草枯对 SH-SY5Y 细胞(100μM, 24 hours) M，24小时)↑ O 欧↑2−• – •production, lipid peroxidation, protein carbonylation, and protein nitration in mitochondrial membranes 线粒体膜中蛋白质的生产，脂质过氧化，蛋白质羰基化和蛋白质硝化
↓ Thiol content in mitochondrial membranes 4. 线粒体膜的硫醇含量
↓ GSH in mitochondria 4. 线粒体中的谷胱甘肽Pinocembrin; 25 μM pretreatment for 4 hours M 预处理4小时↓ O2−• – •production, lipid peroxidation, protein carbonylation, protein nitration, as well as oxidation of thiol groups in mitochondrial membranes 生产，脂质过氧化，蛋白质羰基化，蛋白质硝化，以及线粒体膜巯基的氧化
↑ Thiol content in mitochondrial membranes 线粒体膜中硫醇↑含量
↑ GSH in mitochondrial membranes 线粒体膜中↑型谷胱甘肽
↑ Erk1/2-Nrf2 axis ↑ Erk1/2-Nrf 2轴
↑ GCLM, GCLC, GSH, and HO-1 GCLC、 GSH 和 HO-1Barros Silva et al. [ 巴罗斯 · 席尔瓦等6-OHD-induced dopaminergic neuronal loss in rats, (3 6-ohd 诱导大鼠多巴胺能神经元丢失(3μL, 8 mg/mL, 每公升，8毫克/毫升;s.i我.)↑ Hydrogen peroxide in striatum 纹状体中↑过氧化氢
↑ Cu, Fe, Mn, and Zn in brain 脑中 Cu、 Fe、 Mn、 Zn 的↑Caffeic acid phenethyl ester (CAPE); 10 咖啡酸苯乙酯(CAPE) ;μM/kg, 米/公斤,i.p., cotreatment for 5 days . ，共处理5天↓ Hydrogen peroxide in striatum 在纹状体的↓过氧化氢
↓ Cu, Fe, Mn, and Zn in brain 在大脑中↓铜，铁，锰和锌Mahmoud et al. [ 马哈茂德等人K2CrO4-induced neurotoxicity in rats, (2 mg/kg b.w. for 30 days,目的: 研究 cro4对大鼠神经毒性的影响，探讨 cro4对大鼠神经毒性的作用机制,i.p.)↑ MDA and NO MDA 和 NO
↓ SOD, GPx, and GSH in cerebrum 4. 大脑中的 SOD、 GPx 和 GSH
↑ JAK2, STAT3, and SOCS3 mRNA and protein in cerebrum 大脑中 STAT3、 SOCS3 mRNA 和蛋白的表达CAPE 20 mg/kg b.w. cotreatment for 30 days, orally 咖啡酸苯乙酯20毫克/公斤 b.w. 口服协同治疗30天↓ MDA and NO 4. ↓ MDA 和 NO
↑ SOD, GPx, and GSH in cerebrum 大脑中 SOD、 GPx 和 GSH
↓ JAK2, STAT3, and SOCS3 mRNA and protein in cerebrum 2，STAT3，and SOCS3 mRNA and protein in cerebrumPropolis role in mitigation of chemotherapy side effect 蜂胶在缓解化疗毒副作用中的作用 Kumari et al. [ 库马里等人Mitomycin C-induced testicular toxicity in male mice, (8 mg/kg b.w., 用丝裂霉素 c 诱导雄性小鼠睾丸毒性(8mg/kg b.w,i.p., single dose) . ，单剂量)↑ MDA 温度
↓ GSH, SOD, and CAT in testicular cells 4. 睾丸细胞↓ GSH，SOD 和 CATHydroethanolic extract of Indian propolis pretreatment (1 h prior) 400 mg/kg,印度蜂胶水乙醇提取物预处理(1小时前)400毫克/千克,i.p., single dose . ，单剂量↓ MDA 4. ↓ MDA
↑ GSH and CAT in testicular cells 睾丸细胞中谷胱甘肽和 CATAlyane et al. [等人Doxorubicin-induced toxicity in rat heat mitochondria, (20 mg/kg b.w.,阿霉素对大鼠热线粒体的毒性作用,i.p., single dose) . ，单剂量)↑ Mitochondrial MDA 线粒体
↓ RCR (respiratory chain ratio) and P/O ratio ↓呼吸链比率和 P/O 比率
↑ O 欧↑2−(evaluated(评估in vitro在试管中)Propolis extract pretreatment with 100 mg/kg/day, 蜂胶提取物预处理100毫克/千克/天,p.o.假释官 for four days prior 已经四天了↓ Mitochondrial MDA 4. 名片↓线粒体 MDA
↑ RCR (respiratory chain ratio) and P/O ratio 呼吸链比值与 P/O 比值
↓ O2−(evaluated (评估in vitro 在试管中)Propolis as a modulator of cardiovascular disease markers 蜂胶作为心血管疾病标志物的调节剂 Salmas et al. [ 萨尔马斯等人Nω-nitro-L-arginine methyl ester- (L-NAME-) induced hypertension in rats, (40 mg/kg b.w.; – 硝基 -l- 精氨酸甲酯-(L-NAME -)诱导大鼠高血压(40mg/kg b.w. ;i.p. for 28 days) 为期28天)↓ TAS, PON1 公司名称↓ TAS，PON1
↑ TOS, ADMA, and NF- ↑ TOS、 ADMA 和 NF-κBPropolis CAPE coadministration: propolis: 200 mg/kg/d; 28 days, by gavage; CAPE: 50 蜂胶合剂: 蜂胶: 200毫克/公斤/天; 28天灌胃; CAPE: 50μM/kg/d; 14 days, 每日 m/kg; 14天,i.p.↑ TAS
↑ PON1—only propolis 仅蜂胶
↓ TOS, ADMA4. ↓
↓ NF- ↓-κB—only propolis B ー only 蜂胶Ahmed et al. [ 艾哈迈德等53]Isoproterenol-induced myocardial infarction in rats, (85 mg/kg injection for 2 days—on the 29th and 30th days) 异丙肾上腺素诱导大鼠心肌梗死(85mg/kg 注射2天ー29天和30天)↓ SOD, GPx, GRx, and GST in myocardium 4. 心肌中的 SOD，GPx，GRx 和 GST
↑ TBARS in myocardium 心肌中轻度调节的 TBARSMalaysian propolis ethanol extract, pretreatment with 100 mg/kg/day, orally, 30 days 马来西亚蜂胶乙醇提取物，预处理100mg/kg/d，口服，30d↑ GPx, GRx, and GST in myocardium 心肌中 g↑、 GRx 和 GST
↓ TBARS in myocardium 4. 心肌组织↓ TBARSSun et al. [ 孙等人H2O2-induced rat cardiomyocytes (H9c2) oxidative injury, (700 目的: 探讨大鼠心肌细胞氧化损伤的机制μM, 6 h) 6 h)↑ MDA 温度
↓ SOD and GPx 和 GPxCAPE, benzyl caffeate, and cinnamyl caffeate pretreatment with 1, 5, and 10 咖啡酸苯乙酯、咖啡酸苄酯和咖啡酸肉桂酯预处理1,5,10μM for 12 h 12小时↓ MDA—doses of 5 and 10 5和10的剂量μM
↑ SOD and GPx—doses of 5 and 10 5和10的↑ SOD 和 gpx ー剂量μMEl-Awady et al. [ 埃尔-阿瓦迪等人High glucose-induced vascular endothelial dysfunction, isolated rat aorta, (44 mM for 3 hours) 高糖诱导的血管内皮功能障碍，离体大鼠主动脉(44mm，3小时)↑ TBARS in rat aorta 大鼠主动脉中↑ TBARS
↓ SOD and GSH in rat aorta 4. 大鼠主动脉中的 SOD 和 GSHPropolis extract pretreatment 400 蜂胶提取物预处理400μg/mL, 30 min prior 每克/毫升，30分钟前↓ TBARS in rat aorta 4. 大鼠主动脉的↓ TBARS
↑ SOD and GSH in rat aorta 大鼠主动脉中脑 SOD 和 GSH 的表达Propolis as protective agent against prooxidants’ toxicity 蜂胶作为抗氧化剂的毒性研究 Yonar et al. [ 约纳尔等人Trichlorfon-induced oxidative stress in fish, environmental exposure, 11 and 22 mg/L, 14 days 敌百虫在鱼体内引起的氧化应激，环境暴露量，11和22毫克/升，14天↑ MDA in the liver, kidney, and gill 肝、肾、鳃中的 MDA ↑
↓ GSH, SOD, CAT, and GPx in the liver, kidney and gill 在肝脏、肾脏和鳃的↓ GSH、 SOD、 CAT 和 GPxPropolis cotreatment 10 mg/kg of fish weight, 14 days 蜂胶协同处理10毫克/公斤鱼重，14天↓ MDA in the liver, kidney, and gill 4. 肝脏、肾脏和鳃的 MDA
↑ GSH, SOD, CAT, and GPx in the liver, kidney, and gill 肝、肾、鳃中的 GSH、 SOD、 CAT 和 GPxFerreira et al. [ 费雷拉等人Tebuconazole-induced oxidative stress in fish, environmental exposure (0.88 mg/L) 戊唑醇致鱼体内氧化应激，环境暴露量(0.88毫克/升)↑ MDA and carbonyl protein in brain, liver, and kidney 脑、肝、肾中 MDA ↑和羰基蛋白
↑ GST in liver 肝中 GST ↑
↓ GST in brain 在脑海中↓ GST
↑ CAT in kidney and brain 肾脑中 CAT 的↑
↓ SOD in liver 4. 肝脏里的 SODPropolis; 0.01, 0.05, and 0.1 g/L 蜂胶; 0.01,0.05,0.1克/升↓ MDA and carbonyl protein in brain, liver and kidney 4、大脑、肝脏和肾脏中的 MDA 和羰基蛋白
↑ GST in brain, liver 脑、肝中 GST∗∗ 第 x 次/次and kidney 还有肾脏
↑ CAT in liver 肝脏中的雌雄蕊∗∗ 第 x 次/次
↓ CAT in kidney 肾脏的↓ CAT∗∗ 第 x 次/次 and brain 还有大脑
↑ SOD in liver肝脏中Aksu et al. [ 阿克苏等Paracetamol- (PRC-) induced reproductive toxicity in rats, (500 mg/kg b.w., by oral gavage) 扑热息痛-(PRC -)诱导大鼠生殖毒性(500mg/kg b.w. ，口服灌胃)↓ SOD, CAT, GPx, and GSH in testicular tissue 4、睾丸组织中的 SOD、 CAT、 GPx 和 GSH
↑ MDA in testicular tissue 睾丸组织中 MDA ↑Chrysin; pretreatment with 25 mg/kg and 50 mg/kg b.w., by oral gavage, 7 days 白杨素: 25毫克/千克和50毫克/千克每小时，口服灌胃，7天↑ GSH, CAT 谷胱甘肽∗∗ 第 x 次/次, GPx ，GPx∗∗ 第 x 次/次, SOD (only the higher dose) in testicular tissue 在睾丸组织中，SOD (只有较高剂量)
↓ MDA in testicular tissue 睾丸组织中的 MDAManzolii et al. [ 曼佐利等人Methylmercury-induced oxidative stress (30 甲基汞诱发的氧化应激(30μg/kg b.w., by gavage, 45 days) G/kg b.w，灌胃，45天)↓ GSH in blood ↓血液中的谷胱甘肽Chrysin; cotreatment (0.10, 1.0, and 10 mg/kg b.w., by gavage, 45 days) 白杨素联合处理(0.10,1.0,10mg/kg b.w. 灌胃，45天)↑ GSH in blood 血中谷胱甘肽Saito et al. [ 齐藤等人UVA irradiation, human skin fibroblast cells—NB1-RGB (10 J/cm 长波紫外线照射后，人皮肤成纤维细胞 nb1-rgb (10j/cm2)↑ HO-1 expression ↑ HO-1表达Brazilian green propolis; 3, 10, or 30 巴西绿色蜂胶; 3,10，或30μg/mL 3,5-di- 克/毫升3,5-二 –O-caffeoylquinic acid, 3,4-di- – 咖啡酰奎宁酸3,4-二 –O-caffeoylquinic acid, and chlorogenic acid; 1 or 3 – 咖啡酰奎宁酸和绿原酸;μg/mL 克/毫升↑ HO-1 expression ↑ HO-1表达
↑ Nrf2 nuclear translocation to the nuclei (only propolis extract was studied) 核转位(仅研究了蜂胶提取物)Cao et al. [ 曹等人H2O2-induced oxidative stress, mouse L929 fibroblast cell lines, (600 目的: 研究小鼠 L929成纤维细胞系(600μM H2O, 12 hours) 12小时)↑ ROS 活性氧
↓ Cell viability 4. ↓细胞活力Ethanol extract of Chinese propolis; pretreatment with 5, 7.5, and 10 蜂胶乙醇提取物; 5,7.5,10μg/mL per 3 hours prior 事前3小时每毫升样本含克↓ ROS∗∗ 第 x 次/次
↑ Cell viability ↑细胞活力∗∗ 第 x 次/次
↑ HO-1, GCLM, and GCLC at mRNA level (the highest dose was studied) 在 mRNA 水平上测定了↑ HO-1、 GCLM 和 GCLC (最高剂量)
↑ HO-1 and GCLM at protein level (the highest dose was studied) 在蛋白质水平上，测定了↑ HO-1和 GCLM 的最高剂量Arabameri et al. [ 阿拉伯美里等人Maternal separation-induced stress, the neonatal rats, separated 6 hours per day, 15 days 母体分离诱发应激，新生大鼠每天分离6小时，15天↑ MDA in ovarian tissue 卵巢组织中 MDA ↑
↓ SOD, GPx, and FRAP in ovarian tissue 4. 卵巢组织中的 SOD，GPx 和 FRAPIranian propolis; cotreatment 50, 100, or 200 mg/kg b.w.; 15 days 伊朗蜂胶; 50,100，或200毫克/千克 b.w. ; 15天↓ MDA 4. ↓ MDA∗∗ 第 x 次/次in ovarian tissue 在卵巢组织中
↑ SOD∗∗ 第 x 次/次, GPx, and FRAP 和 FRAP∗∗ 第 x 次/次 in ovarian tissue 在卵巢组织中
All three doses exerted a positive effect, but the most effective was 200 mg/kg 所有三种剂量都有积极作用，但最有效的是200毫克/千克Zhang et al. [张等人H2O2-induced oxidative stress, RAW264.7 cells, 300 – 诱导氧化应激，RAW264.7细胞，300μM for 13 hours 13小时↑ Intracellular ROS 胞内↑活性氧Two ethanol extracts of Chinese propolis, pretreatment for 0.5 hour before 两种中国蜂胶乙醇提取物，预处理0.5小时前↓ Intracellular ROS 4. 细胞内的 ROSRAW264.7 cells not subjected to any factor RAW264.7细胞不受任何因素影响———— ーーTwo ethanol extracts of Chinese propolis 中国蜂胶的两种乙醇提取物↓ Intracellular ROS 4. 细胞内的 ROS
↑ HO-1, GCLM, and TrxR1 on both the mRNA 结果表明，mRNA 上存在↑ HO-1、 GCLM 和 TrxR1，而 mRNA 上存在↑ HO-1、 GCLM 和 TrxR1∗∗ 第 x 次/次 and protein levels 和蛋白质水平∗∗ 第 x 次/次
The most effective for HO-1 对 HO-1最有效的
|ADMA: asymmetric dimethylarginine; Bax: Bcl-2-related ovarian killer protein; Bcl-2: B-cell lymphoma 2; CAT: catalase; GCLC: glutamate-cysteine ligase catalytic subunit; GCLM: glutamate-cysteine ligase regulatory subunit; Erk 1/2: extracellular signal-regulated kinase ½, FRAP: ferric reducing ability; GPx: glutathione peroxidase; GRx: glutathione reductase; GSH: reduced glutathione; GST: glutathione reductase; HO-1: heme oxygenase-1; JAK 2: Janus kinase 2; MDA: malondialdehyde; NF- ADMA: 非对称性二甲基精氨酸; Bax: Bcl-2相关的卵巢杀伤蛋白; Bcl-2: b 细胞淋巴瘤2; CAT: 过氧化氢酶; GCLC: 谷氨酸半胱氨酸连接酶催化亚单位; GCLM: 谷氨酸半胱氨酸连接酶调节亚单位; Erk 1/2: 细胞外信号调节激酶12，FRAP: 铁还原能力; GPx: 谷胱甘肽过氧化物酶; GRx: 谷胱甘肽还原酶; GSH: 谷胱甘肽; GST: GST: ; HO-1: 血红素加氧酶1; JAK 2: Janus 2; MDA: 丙二醛; NF-κB: nuclear factor kappa B; Nrf2: nuclear factor erythroid 2-related factor 2 (Nrf2); NO: nitric oxide; 8-oxo-2′-deoxyguanosine, P/O: phosphate/oxygen ratio; PON1: paraoxonase; RCR: respiratory control ratio; ROS: reactive oxygen species; STAT3: signal transducer and activator of transcription 3; SOCS3: suppressor of cytokine signaling 3; SOD: superoxide dismutase; TAS: total antioxidant status; TBARS: thiobarbituric acid reactive substances; TOS: total oxidant status; TrxR1: thioredoxin reductase 1; B: 核因子 kappa b; Nrf2: 核因子红系统2相关因子2(Nrf2) ; NO: 一氧化氮; 8-oxo-2′-脱氧鸟苷，P/O: 磷酸/氧比; PON1: 对氧磷酶; RCR: 呼吸控制比; ROS: 活性氧类; STAT3: STAT3; SOCS3: 细胞因子信号抑制因子3; 抗氧化剂: 超氧化物歧化酶; TAS: 总状态; TBARS: 硫代巴比妥酸物质; TOS: 总氧化状态; TrxR1: 硫氧还蛋白还原酶1;γ-GCS: γ γ — gc: γ-glutamylcysteine synthetase. ↓: decrease; ↑: increase; _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _∗∗ 第 x 次/次the effect depended on used dose. 效果取决于使用剂量|
Table 2 表二The protective effects of propolis against prooxidant action of different harmful factors. 蜂胶对不同有害因子的抗氧化作用
Jin 等人 ，Barros Silva 等人和 de Oliveira 等人研究了蜂胶及其化合物神经保护作用的潜在机制。
Jin et al.  reported that pinocembrin, one of the most abundant flavonoids in propolis, inhibited 6-hydroxydopamine- (6-OHDA-) induced oxidative stress. Pinocembrin pretreatment induced the translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) to the nucleus in a concentration- and time-dependent manner as well as the subsequent expression of antioxidant response element- (ARE-) mediated antioxidant genes encoding heme oxygenase-1 (HO-1) and γ-glutamylcysteine synthetase (γ-GCS). Nrf2 is known to play a key role in the adaptive response to oxidative and electrophilic stresses as well as maintaining the cellular self-defense. Under physiological condition, Nrf2 is localized in the cytosol and is associated with its negative regulator, Kelch-like ECH-associated protein 1 (Keap1). In response to oxidative/electrophilic stimuli, Nrf2 dissociates from Keap1 and translocates to the nucleus, where it forms a heterodimer with its obligatory partner Maf and then binds to the ARE sequence to activate transcription of genes encoding a large number of antioxidative and electrophile detoxification enzymes including HO-1 and γ-GCS. Pinocembrin also reduced the 6-OHDA-induced cell viability loss and apoptotic rate and partially inhibited the reduction of the Bcl-2 (an apoptosis inhibitor) to Bax (an apoptosis promoter) ratio following 6-OHDA treatment. The treatment of SH-SY5Y cells with the small interfering RNA (siRNA) directed against Nrf2 (Nrf2-siRNA) abolished pinocembrin-induced HO-1 and γ-GCS expression and its protective effects, which suggests that pinocembrin is protective against Parkinson’s disease-related neurotoxin 6-OHDA through Nrf2/ARE pathway (Table 2). De Oliveira et al.  confirmed that pinocembrin exerted mitochondrial and cellular protection by the activation of the extracellular signal-regulated kinase 1/2-nuclear factor erythroid 2-related factor (Erk1/2-Nrf2) signaling pathway, since the inhibition of Erk1/2 or the silencing of Nrf2 abrogated these effects. Erk1/2 protein kinase is an activator of Nrf2. The researchers showed that pinocembrin pretreatment inhibited paraquat-induced lipid peroxidation, protein carbonylation, protein nitration, as well as the oxidation of thiol groups in the membranes of mitochondria of SH-SY5Y cells. Moreover, it activated the translocation of Nrf2 and increased the level of glutamate-cysteine ligase regulatory subunit (GCLM), glutamate-cysteine ligase catalytic subunit (GCLC), GSH, and HO-1. GCLM and GCLC are, respectively, regulatory and catalytic subunits of glutamate cysteine ligase—an enzyme which catalyzes the first and rate-limiting step in the production of the cellular antioxidant GSH. The above effects were blocked or inhibited with the Erk1/2 protein kinase inhibitor PD98059 or Nrf2 siRNA (Table 2).
据 Jin 等人报道，松黄素是蜂胶中含量最丰富的黄酮类化合物之一，可以抑制6- 羟基多巴胺(6-OHDA -)诱导的氧化应激。松黄素预处理诱导红系核因子2相关因子2(Nrf2)以浓度和时间依赖的方式向细胞核移位，并诱导抗氧化反应元件-(ARE -)介导的抗氧化基因编码血红素氧合酶 -1(HO-1)和 γ- 谷氨酰半胱氨酸合成酶(γ-gcs)的表达。已知 Nrf2在对氧化和亲电应激的适应性反应以及维持细胞的自我防御中起着关键作用。在生理状况下，Nrf2定位于细胞溶胶中，并与其负调节因子 Kelch-like ech- associated protein 1(Keap1)相关。在亲性氧化刺激的作用下，Nrf2从 Keap1中分离出来，转移到细胞核，与其结合位点 Maf 形成异二聚体，结合 ARE 序列激活编码大量抗氧化和亲电解毒酶的基因转录，包括 HO-1和 γ-gcs。Pinocemrin 还能降低6-OHDA 诱导的细胞活力损失和凋亡率，并部分抑制6-OHDA 处理后 Bcl-2(凋亡抑制剂)降低为 Bax (凋亡促进剂)的比例。用针对 Nrf2(Nrf2-siRNA)阻断 pinocembrin 诱导的 HO-1和 γ-gcs 表达的 siRNA 治疗 SH-SY5Y 细胞及其保护作用，提示 pinocembrin 通过 Nrf2/ARE 途径对帕金森病相关神经毒素6-OHDA 具有保护作用(表2)。De Oliveira 等人证实松果体素通过激活细胞外信号调节激酶1/2核因子红系因子2相关因子(Erk1/2-Nrf 2)信号通路而发挥线粒体和细胞保护作用，因为 Erk1/2的抑制或 Nrf2的沉默消除了这些作用。Erk1/2蛋白激酶是 Nrf2的激活剂。研究人员表明，pinocembrin 预处理抑制百草枯诱导的脂质过氧化、蛋白质羰基化、蛋白质硝化，以及 SH-SY5Y 细胞线粒体膜上巯基的氧化。此外，它还能激活 Nrf2的转位，提高谷氨酸半胱氨酸连接酶调节亚基(GCLM)、谷氨酸半胱氨酸连接酶催化亚基(GCLC)、谷胱甘肽(GSH)和 HO-1的水平。GCLM 和 GCLC 分别是谷氨酸半胱氨酸连接酶的调节亚基和催化亚基，谷氨酸半胱氨酸连接酶是催化细胞抗氧化剂 GSH 生成的第一步和限速步骤。上述作用被 Erk1/2蛋白激酶抑制剂 PD98059或 Nrf2 siRNA 阻断或抑制(表2)。
The neuroprotective effect of another compound abundant in propolis, namely, caffeic acid phenethyl ester (CAPE), against 6-OHDA-induced dopaminergic neuronal loss in rats, was studied by Barros Silva et al. . The cotreatment with CAPE decreased the hydrogen peroxide production in brain striatum homogenates. CAPE was also capable of scavenging ROS by neutralizing the unpaired electrons of DPPH but did not affect 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL, a stable nitroxyl antioxidant) in brain-affected areas. Additionally, CAPE protected against 6-OHDA-induced increase of metal levels (Cu, Fe, Mn, and Zn) as well as inhibited mitochondrial permeability transition (MPT), a mediator of neuronal death that triggers cytochrome c release and caspase-3 activation, and this effect was not associated with mitochondrial dysfunction. The authors concluded that basing on the obtained findings and its ability to cross the blood-brain barrier, CAPE could be a promising compound to treat Parkinson’s and other neurodegenerative diseases (Table 2). Mahmoud et al.  demonstrated, in turn, that CAPE protected the brain against hexavalent chromium toxicity by preventing oxidative/nitrosative stress as well as modulating the JAK/STAT signaling pathway in rats. The researchers suggested that oxidative stress along with inflammation caused by Cr(VI) could directly activate the JAK/STAT signaling pathway in the cerebrum of rats, which was confirmed by increased JAK2 mRNA and protein expression and consequently STAT3 mRNA and protein phosphorylation in the cerebrum of Cr(VI)-induced rats. CAPE, in turn, by mitigating oxidative/nitrosative stress, downregulated JAK2/STAT3 signaling, which was also proved by a significant decrease in both JAK2 and STAT3 mRNA and protein levels in CAPE-treated group (Table 2).
巴罗斯 · 席尔瓦等人研究了蜂胶中含量丰富的咖啡酸苯乙酯(CAPE)对6-ohda 所致大鼠多巴胺能神经元损伤的保护作用。咖啡酸苯乙酯联合处理降低了大脑纹状体匀浆中过氧化氢的产生。CAPE 也能通过中和 DPPH 的未配对电子清除活性氧，但不影响大脑受影响区域的4- 羟基 -2,2,6,6- 四甲基哌啶 -n- 氧基(TEMPOL，一种稳定的硝基抗氧化剂)。此外，CAPE 对6-ohda 诱导的金属水平(Cu，Fe，Mn，Zn)升高具有保护作用，并抑制了线粒体通透性转换(MPT) ，这是一种引起细胞色素 c 释放和 caspase-3激活的神经元死亡介质，这种作用与线粒体功能障碍无关。作者得出结论，基于所获得的研究结果及其穿越血脑屏障的能力，CAPE 可能是一种有前途的化合物，用于治疗帕金森氏症和其他神经退行性疾病(表2)。Mahmoud et al. 反过来证明，CAPE 通过防止氧化/硝化应激和调节大鼠 JAK/STAT 信号通路，保护大脑免受六价铬中毒的影响。研究人员提出，氧化应激和 Cr (VI)引起的炎症可以直接激活大鼠大脑中 JAK/STAT 信号通路，这一点可以通过增加 JAK2 mRNA 和蛋白的表达以及随后在 Cr (VI)诱导的大鼠大脑中 STAT3 mRNA 和蛋白磷酸化得到证实。CAPE 依次通过减轻氧化/硝化应激，下调 JAK2/STAT3信号转导，这也被证明是通过明显降低 JAK2和 STAT3 mRNA 和蛋白水平在 CAPE 治疗组(表2)。
2.5. Propolis Role in Mitigation of Chemotherapy Side Effect
In the literature, there are also studies aiming to determine propolis as a potential natural antioxidant to mitigate side effect of chemotherapy. Mitomycin C, cisplatin, and doxorubicin are recognised anticancer drugs used along with radiation or surgery. Unfortunately, their administration may cause diverse side effects, leading to considerable injury of organs and subsequently worsening life conditions. Some aspects of this harmful effect have been attributed to inducing oxidative damage.
Kumari et al.  showed that the hydroethanolic extract of Indian propolis (HEIP) displayed the protective effect against mitomycin C- (MMC-) induced genotoxicity and cytotoxicity which could be, at least partially, mediated via free radical-scavenging activity and inhibitory effect on lipid peroxidation. The potential geno- and cytotoxic effects of MMC in the bone marrow was manifested by a significant increase in the frequency of micronculeated cells and the percentage of apoptotic cells as well as the reduction in polychromatic erythrocyte (PCE) to normochromatic erythrocytes (NCE) ratio (P/N ratio) compared to the control group. However, MMC-induced toxic effects were significantly recovered by pretreatment with HEIP with the optimum dose being 400 mg/kg. In addition, HEIP possessed a considerable DPPH radical-scavenging activity (increasing along with an increase in HEIP concentration), and it exhibited almost such effectiveness as the standard use of ascorbic acid. HEIP was also found to possess the total antioxidant activity (evaluated by method based on the principle of reduction of molybdenum) and exhibited free radical-scavenging activity in FRAP measurement, but in this case, the results were not comparable with those obtained for ascorbic acid. Moreover, HEIP was shown to possess a substantial lipid peroxidation inhibitory activity, but again, it was also not as effective as the reference standard—Trolox (Table 2).
表明印度蜂胶的水乙醇提取物对丝裂霉素 c-(MMC -)诱导的遗传毒性和细胞毒性具有保护作用，至少部分通过自由基清除活性和脂质过氧化抑制作用介导。与对照组相比，MMC 在骨髓中的潜在基因和细胞毒性作用表现为显著增加微克隆细胞的频率和凋亡细胞的百分比，以及降低多染红细胞与正常红细胞的比率(P/N)。最佳剂量为400mg/kg 时，预处理能显著恢复 mmc 的毒性效应。此外，HEIP 还具有较强的 DPPH 自由基清除活性(随着 HEIP 浓度的增加而增加) ，其清除活性与抗坏血酸的标准用量相当。HEIP 还具有总的抗氧化活性(以还原钼原理为基础的方法进行评价) ，在 FRAP 测定中表现出自由基清除活性，但在这种情况下，其结果与抗坏血酸无可比性。此外，HEIP 被证明具有明显的脂质过氧化抑制活性，但是同样，它也没有参考标准曲洛克斯有效(表2)。
As chemotherapy is known to have fertility-related side effects, in the next study, Kumari et al.  investigated the effect of HEIP on MMC-induced testicular toxicity. The antioxidant effects of HEIP were assessed by measuring antioxidant/oxidant biomarkers in testicular tissue homogenate. MMC treatment resulted in long-term oxidative stress, whereas a single dose preadministration of HEIP was able to attenuate it to a certain degree—a significant decrease in MDA level and an insignificant elevation of GSH level and CAT activity were observed. MMC administration also led to a reduction in testicular function (testis weight, sperm count, sperm motility, and spermatozoa with normal head morphology) in a dose-dependent manner, which was alleviated by HEIP pretreatment (Table 2).
由于化疗已知有生育相关的副作用，在下一个研究中，Kumari 等人调查了 HEIP 对 mmc 引起的睾丸毒性的影响。通过测定睾丸组织匀浆中的抗氧化/氧化生物标志物评价 HEIP 的抗氧化作用。丝裂霉素处理可导致长期的氧化应激，而单剂量的 HEIP 预处理能够在一定程度上减弱，MDA 水平显著降低，GSH 水平和 CAT 活性升高不明显。丝裂霉素还导致睾丸功能(睾丸重量，精子计数，精子活力和头部形态正常的精子)的剂量依赖性减少，这是通过帮助预处理减轻(表2)。
Alyane et al. , in turn, demonstrated that the propolis extract pretreatment substantially attenuated the peroxidative damage in the heart mitochondria following the injection of an acute dose of doxorubicin. Propolis led to a significant reduction in mitochondrial MDA formation and production of superoxide anion, as well as the restoration of respiratory control ratio (RCR—state III respiration/state IV respiration; indicates the tightness of the coupling between respiration and phosphorylation) and phosphate/oxygen ratio (P/O ratio; refers to the amount of ATP produced from the movement of two electrons through a defined electron transport chain. Additionally, a decreased rate and amplitude of mitochondrial swelling were observed (Table 2).
反过来，Alyane 等人证明蜂胶提取物预处理大大减轻了急性剂量阿霉素注射后心脏线粒体的过氧化损伤。蜂胶导致线粒体 MDA 的形成和超氧阴离子的产生显著减少，以及呼吸控制比率的恢复(rcr ー state III respiration/state IV respiration; 表明呼吸和磷酸化之间的紧密耦合)和磷酸盐/氧比率(P/O 比率; 指的是通过一定的电子传递链运动产生的 ATP 的数量。此外，减少率和振幅的线粒体肿胀观察到(表2)。
2.6. Propolis’ Ability to Modulate Cardiovascular Disease Markers
The antioxidant properties of propolis have also been suggested to be responsible for its ability to modulate cardiovascular disease markers. Salmas et al. , for example, reported that oxidative alterations occurring in the kidney tissue of chronic hypertensive rats might be prevented via propolis, CAPE, as well as pollen administration. In the kidney tissue of Nω-nitro-L-arginine methyl ester- (L-NAME-) induced hypertensive rats, total antioxidant status (TAS) and paraoxonase (PON1, an important antioxidant preventing the oxidation of low-density lipoproteins) activity, were significantly decreased, whereas total oxidant status (TOS), asymmetric dimethylarginine (ADMA, an endogenous inhibitor of NO synthase), and nuclear factor kappa B (NF-κB, regulated by intracellular redox state) were significantly increased. However, the coadministration of propolis, CAPE, and pollen restored all the disturbed parameters with the propolis samples being the most effective following by pollen and CAPE (Table 2).
蜂胶的抗氧化特性也被认为与其调节心血管疾病标志物的能力有关。例如，Salmas et al. 报告说，慢性高血压大鼠肾组织中发生的氧化改变可以通过蜂胶、 CAPE 和花粉给药来预防。在 Nω-nitro-L-arginine 甲酯(L-NAME)诱导的高血压大鼠肾组织中，总抗氧化状态(TAS)和对氧磷酶(PON1，一种防止低密度脂蛋白氧化的重要抗氧化剂)活性显著降低，而总氧化状态(TOS)、非对称性二甲基精氨酸(ADMA，一氧化氮合酶的内源抑制剂)和核转录因子 κb (NF-κB，由氧化还原状态调节)则显著升高。然而，蜂胶、 CAPE 和花粉的共同作用恢复了所有的干扰参数，蜂胶样品是最有效的，其次是花粉和 CAPE (表2)。
Ahmed et al. , in turn, showed that Malaysian propolis (MP) pretreatment ameliorated the negative effects of isoproterenol-induced myocardial infarction in rats. MP exhibited high total antioxidant activity determined by both DPPH and FRAP assays. The isoproterenol administration resulted in significantly elevated lipid peroxides and reduced activities of cellular antioxidant defense enzymes in the myocardium. Moreover, it caused a significant increase in serum cardiac marker enzymes (creatinine kinase-MB, aspartate transaminase, lactate dehydrogenase, and alanine transaminase) and cardiac troponin I levels as well as altered serum lipid profiles. However, the pretreatment of ischemic rats with MP suppressed the above biochemical parameters as well as improved histopathological findings, suggesting the protective effect of MP against ISO-induced ischemia via its direct cytotoxic radical-scavenging activities and possibly via the inhibition of lipid peroxidation (Table 2).
Ahmed 等反过来指出，马来西亚蜂胶(MP)预处理可以改善异丙肾上腺素诱导的心肌梗死对大鼠的负面影响。DPPH 法和 FRAP 法均显示 MP 具有较高的总抗氧化活性。服用异丙肾上腺素后，心肌组织脂质过氧化物明显升高，细胞抗氧化防御酶活性降低。此外，它还引起血清心肌标记酶(肌酐激酶-mb、天冬氨酸氨基转移酶、乳酸脱氢酶和谷丙转氨酶)和心肌肌钙蛋白 i 水平的显著增加以及血清脂质谱的改变。然而，MP 预处理缺血大鼠抑制上述生化参数以及改善组织病理学结果，提示 MP 对 iso 诱导的缺血的保护作用是通过其直接的细胞毒性自由基清除活性和可能通过抑制脂质过氧化的作用实现的(表2)。
The protective effect of six active compounds of Chinese propolis on H2O2-induced rat cardiomyocytes (H9c2) oxidative injury was also studied . All tested compounds demonstrated significant cytoprotective activities; however CAPE, benzyl caffeate (BZC), and cinnamyl caffeate (CNC) exerted stronger effects than chrysin, pinobanksin, and 3,4-dimethoxycinnamic acid (DMCA). CAPE, BZC, and CHC increased H9c2 cellular antioxidant potential (by decreasing MDA level and increasing SOD and GPx activities), decreased intracellular calcium ion level, and prevented cell apoptosis (Table 2).
本文还研究了六种蜂胶活性成分对 h2o2诱导的大鼠心肌细胞氧化损伤的保护作用。所有化合物均具有明显的细胞保护活性，咖啡酸苄酯(BZC)和咖啡酸肉桂酯(CNC)的细胞保护活性均强于白杨素、品蓝素和3,4- 二甲氧基肉桂酸(DMCA)。CAPE、 BZC 和 CHC 增加 H9c2细胞的抗氧化能力(通过降低 MDA 水平、增加 SOD 和 GPx 活性) ，降低细胞内钙离子水平，阻止细胞凋亡(表2)。
The protective effects of ethanol extract of propolis (EEP) against injury induced by oxidized low-density lipoprotein (ox-LDL) in human umbilical vein endothelial cells (HUVECs) were studied by Fang et al. . An atherogenic role of ox-LDL in the progression of atherosclerotic cardiovascular disease is well known. EEP pretreatment ameliorated the ox-LDL-induced oxidative stress by reducing nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation, ROS, and MDA generation as well as elevating antioxidant enzyme activities. In addition, EEP reduced ox-LDL uptake by HUVECs and attenuated ox-LDL-upregulated expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1—a critical molecule responsible for ox-LDL uptake by endothelial cells) both at the mRNA and at protein levels . Moreover, EEP in a dose-dependent manner protected against the decrease in cell viability as well as the increase in lactate dehydrogenase (LDH) release, caspase-3 activation, and apoptosis induced by ox-LDL. The obtained outcomes allowed the authors to conclude that EEP appeared to protect HUVECs from ox-LDL-induced injury via, at least partially, the modulation of LOX-1-mediated oxidative stress.
方等研究了蜂胶乙醇提取物(EEP)对人脐静脉内皮细胞(HUVECs)氧化修饰低密度脂蛋白(ox-LDL)损伤的保护作用。牛低密度脂蛋白在冠状动脉疾病增生过程中的致动脉粥样硬化作用是众所周知的。EEP 预处理通过降低 NADPH 氧化酶的活性、活性氧和丙二醛的产生以及提高抗氧化酶的活性，改善氧化低密度脂蛋白(ox-ldl)诱导的氧化应激。除此之外，EEP 还可以减少 HUVECs 对 ox-LDL 的摄取，并在 mRNA 和蛋白水平上减弱凝集素样氧化低密度脂蛋白受体1(lox-1，内皮细胞摄取 ox-LDL 的关键分子)的表达。此外，EEP 以剂量依赖的方式保护细胞活力下降，以及乳酸脱氢酶释放增加，caspase-3激活和氧化低密度脂蛋白诱导的凋亡。所获得的结果使作者得出结论: EEP 似乎可以保护 HUVECs 免受牛低密度脂蛋白所致的损伤，至少部分是通过调节 lox-1介导的氧化应激。
Tian et al.  proved that the ethanol extract of propolis might protect macrophages from ox-LDL-induced apoptosis and the underlying mechanism at least partially involved its ability to suppress the CD36-mediated ox-LDL intake and the subsequent activation of the endoplasmic reticulum (ER) stress-C/EBP homologous protein (CHOP) pathway; it significantly suppressed the phosphorylation of double-stranded RNA-activated protein kinase-like ER kinase (PERK) and eukaryotic translation initiation factor 2α (eIF2α) as well as the upregulation of glucose regulated protein 78 (GRP78) and the proapoptotic protein CHOP.
甜等证明蜂胶乙醇提取物可能保护巨噬细胞免受牛低密度脂蛋白(ox-LDL)诱导的细胞凋亡，其机制至少部分涉及其抑制 cd36介导的牛低密度脂蛋白(ox-LDL)摄入和随后激活的内质网应激 c/ebp 同源蛋白(CHOP)途径，它显著抑制双链 rna 激活的蛋白激酶样 ER 激酶(PERK)和真核翻译起始因子2 α (eif2α)的磷酸化，以及调节的葡萄糖78(GRP78)和凋亡蛋白 CHOP 的上调。
El-Awady et al. , in turn, reported that propolis could protect against high glucose-induced vascular endothelial dysfunction via decreasing oxidative stress in isolated rat aorta. Incubation of aortic rings with propolis extract prevented high glucose-induced the impairment of phenylephrine-induced contraction and acetylcholine-induced relaxation. Additionally, a SOD activity and GSH concentration increase as well as a decrease in MDA level were observed (Table 2).
El-Awady et al. 反过来报道，蜂胶可以通过降低离体大鼠主动脉的氧化应激来保护高糖诱导的血管内皮功能障碍。蜂胶提取物与主动脉环孵育预防高糖诱导的苯肾上腺素收缩和乙酰胆碱松弛损伤。此外，超氧化物歧化酶(SOD)活性和谷胱甘肽(GSH)浓度增加，丙二醛(MDA)水平下降(表2)。
2.7. Propolis as the Protective Agent against Prooxidants’ Toxicity
Antioxidant properties of propolis encouraged the research concerning its application as an agent preventing or alleviating harmful oxidative processes caused by various factors, like trichlorfon, tebuconazole, paracetamol, methylmercury, or UV irradiation.
The beneficial effect of propolis on trichlorfon-induced prooxidant/antioxidant and haematological parameters alterations in carp Cyprinus carpio was stated . Fish were exposed to sublethal concentrations of trichlorfon—a toxic pesticide commonly used in aquaculture to eliminate fish parasites, and propolis was administered simultaneously. The treatment with propolis caused the alleviation of trichlorfon-induced negative alterations in the haematological parameters (red and white blood cell counts, haemoglobin concentration, haematocrit, erythrocyte indices, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration) as well as oxidant markers (MDA, GSH, SOD, CAT, and GPx) in the liver, kidney, and gill samples (Table 2). Ferreira et al. , in turn, showed that bee products, such as propolis, bee pollen, royal jelly, and honey, prevented and/or reversed tissue (brain, liver, and kidney) oxidative damage induced by tebuconazole (an agrochemical fungicide) in fish by increasing the enzymatic activities of SOD, CAT, and glutathione-S-transferase (GST) and decreasing lipid peroxidation (Table 2).
研究了蜂胶对敌百虫诱导的鲤鱼抗氧化剂和血液生理指标的影响。将鱼暴露于亚致死浓度的敌百虫中，同时给予蜂胶。敌百虫是水产养殖中常用的杀虫剂，用于消灭鱼类寄生虫。蜂胶治疗可以减轻敌百虫引起的血液学参数(红细胞和白细胞计数、血红蛋白浓度、红细胞压积、红细胞指数、平均红细胞体积、红细胞平均血红蛋白、红细胞平均血红蛋白浓度)以及肝脏、肾脏和鳃标本中氧化标记物(MDA、 GSH、 SOD、 CAT 和 GPx)的负性改变(表2)。费雷拉等人反过来证明，蜂产品，如蜂胶、蜂花粉、蜂王浆和蜂蜜，通过提高 SOD、 CAT 和谷胱甘肽 s- 转移酶(GST)的酶活性和降低脂质过氧化，防止和/或逆转戊唑醇(一种农用化学杀菌剂)在鱼体内诱导的组织(脑、肝和肾)氧化损伤。
Aksu et al.  investigated the effect of chrysin (CR, a flavonoid occurring in propolis) pretreatment against paracetamol- (PRC-) induced reproductive toxicity in male. The treatment with PRC resulted in reduced sperm motility, antioxidant enzymes activity (SOD, CAT, and GPx), and GSH level, as well as increased dead sperm rate, abnormal sperm cell rate, apoptosis, and MDA level in testicular tissue. CR was found to mitigate the above effects in a dose-dependent manner with the higher dose being more effective. The authors concluded that the possible protection mechanism might be dependent on the antioxidant activity of CR (Table 2). A protective effect of chrysin against methylmercury-induced genotoxicity and oxidative stress was also studied by Manzolli et al. . The cotreatment with chrysin resulted in the restoration of GSH level, and it decreased the formation of comets in leukocytes and hepatocytes (Table 2).
阿克苏等人研究了对乙酰氨基酚(PRC -)诱导雄性生殖毒性的白杨素(CR，蜂胶中的一种类黄酮)预处理效果。PRC 治疗后精子活力、抗氧化酶活性(SOD、 CAT、 GPx)和 GSH 水平降低，死精率、精子异常率、细胞凋亡和睾丸组织 MDA 水平升高。发现 CR 以剂量依赖的方式减轻上述影响，高剂量更有效。作者的结论是，可能的保护机制可能依赖于 CR 的抗氧化活性(表2)。Manzolli 等人还研究了白杨素对甲基汞引起的遗传毒性和氧化应激的保护作用。与白杨素共同处理可以恢复 GSH 水平，并减少白细胞和肝细胞中彗星的形成(表2)。
Saito et al. , in turn, demonstrated that Brazilian green propolis and its three main constituents (3,5-di-O-caffeoylquinic acid, 3,4-di-O-caffeoylquinic acid, and chlorogenic acid) increased the HO-1 (heme oxygenase 1) expression and accelerated Nrf2 nuclear translocation after UVA irradiation (the major cause of human skin aging) in human skin fibroblast cells (NB1-RGB).
Saito et al. 进一步证明，UVA 照射后，巴西绿蜂胶及其3种主要成分(3,5-二 o- 咖啡酰奎宁酸、3,4-二 o- 咖啡酰奎宁酸和绿原酸)增加了人皮肤成纤维细胞(NB1-RGB)中 HO-1(HMOX1)的表达，加速了 Nrf2核转位(人类皮肤老化的主要原因)。
Propolis has also been suggested to be useful for improving wound healing, which is possibly owed to its antioxidant activity (Table 2). Cao et al.  investigated the protective effects of the ethanol extract of Chinese propolis (EECP) against H2O2-induced oxidative stress in mouse L929 fibroblast cell lines. EECP not only showed significant protective effects against H2O2-stimulated L929 cell death but also reduced the decline of collagen mRNA expression in a significant way. Moreover, EECP induced the expression of antioxidant-related genes, such as HO-1 (encoding heme oxygenase 1), GCLM(encoding glutamate-cysteine ligase regulatory subunit), and GCLC (encoding glutamate-cysteine ligase catalytic subunit) at both mRNA and protein levels in skin fibroblasts (Table 2). Heme oxygenase-1 breaks down heme to carbon monoxide, iron ions, and biliverdin, which is subsequently reduced to bilirubin. Both biliverdin and bilirubin are potent antioxidant agents.
蜂胶也被认为是有用的，以改善伤口愈合，这可能是由于其抗氧化活性(表2)。曹等研究了蜂胶乙醇提取物(EECP)对 h2o2诱导的小鼠 L929成纤维细胞氧化应激的保护作用。EECP 不仅对 h2o2诱导的 L929细胞死亡有明显的保护作用，而且对胶原 mRNA 表达的下降也有明显的抑制作用。此外，EECP 还在皮肤成纤维细胞的 mRNA 和蛋白水平上诱导抗氧化相关基因的表达，如 HO-1(编码 HMOX1)、 GCLM (编码谷氨酸半胱氨酸结合酶调节亚基)和 GCLC (编码谷氨酸半胱氨酸结合酶催化亚基)。血红素加氧酶 -1将血红素分解为一氧化碳、铁离子和胆绿素，后者随后还原为胆红素。胆绿素和胆红素都是有效的抗氧化剂。
Arabameri et al.  proved that the Iranian propolis could significantly prevent oxidative stress (by alleviating the changes in ferric-reducing antioxidant power, SOD, GPx, and MDA) as well as histopathological changes (the number of ovarian follicles, oocytes, and oocytes diameter) in the ovaries of the neonatal rat following maternal separation stress (infants were separated from their mothers 6 hours per day). All three applied doses exerted positive effect, but the most effective was the dose of 200 mg/kg (Table 2). Mişe Yonar et al. , in turn, investigated the effect of dietary propolis on the number and size of pleopodal egg and oxidative/antioxidant status of freshwater crayfish. Dietary propolis supplementation resulted in a significant decrease in MDA level and CAT and GPx activities as well as a significant increase in SOD activity in hepatopancreas and ovarium. The pleopodal egg number produced per gram of the body weight and total pleopodal egg number significantly increased, whereas the pleopodal egg size significantly decreased following propolis administration. The authors suggested that the reduced activity of CAT and GSH-Px following propolis supplementation could have resulted from the inhibition of superoxide radical formation by the dietary propolis and concluded that propolis improved efficiency in the crayfish and reduced the oxidative stress under controlled hatchery conditions.
阿拉伯美里等人证明，伊朗蜂胶可以明显阻止母体分离应激后新生大鼠卵巢中的氧化应激(通过减轻还原铁的抗氧化能力、 SOD、 GPx 和 MDA 的变化)以及组织病理学变化(卵泡数量、卵母细胞数量和卵母细胞直径)。所有三种剂量都产生了积极的效果，但最有效的剂量是200毫克/千克(表2)。接着，研究了日粮蜂胶对多毛蛋数量和大小以及淡水小龙虾氧化/抗氧化状态的影响。结果表明，蜂胶能显著降低肝胰腺和卵巢组织中 MDA 含量、 CAT 和 GPx 活性，显著提高 SOD 活性。蜂胶使每克体重产生的胸膜卵数和总胸膜卵数显著增加，而使胸膜卵大小显著减小。作者认为，蜂胶补充后 CAT 和 GSH-Px 活性的降低可能是由于蜂胶抑制了食物中超氧化物自由基的形成，并得出结论: 在控制孵化条件下，蜂胶提高了螯虾的效率，降低了氧化应激。
Zhang et al.  demonstrated that ethanol extracts of Chinese propolis (EECP) could reduce the intracellular ROS level not only in the H2O2-induced RAW264.7 cells but also in normal RAW264.7 cell (not subjected to any factor). This suggested that propolis could be capable of reducing oxidative stress generated not only under pathological but also under physiological conditions. Similar to Cao et al. , the authors found that EECP in a time- and dose-dependent manner elevated the expression of antioxidant genes such as HO-1, GCLM, and thioredoxin reductase 1 (TrxR1) on both the mRNA and protein levels (Table 2). TrxR, along with NADPH and thioredoxin, is a component of thioredoxin (Trx) system that creates a key antioxidant system as defense against oxidative stress through the disulfide reductase activity, regulating protein dithiol/disulfide balance . Since EECP also increased expressions of phosphorylated Erk and the nucleus translocation of Nrf2, the researchers suggested that propolis might modulate the expression of HO-1, TrxR1, and GCLM via Erk kinase/Nrf2 signal pathway.
张等证明中国蜂胶乙醇提取物(EECP)不仅能降低 h2o2诱导的 RAW264.7细胞内 ROS 水平，而且能降低正常 RAW264.7细胞内 ROS 水平(不受任何因素影响)。这表明蜂胶不仅可以降低病理条件下产生的氧化应激，而且可以降低生理条件下产生的胆固醇。与 Cao et al. 相似，作者发现 EECP 以一种时间和剂量依赖的方式提高了 mRNA 和蛋白水平上的抗氧化基因的表达，如 HO-1，GCLM 和硫氧还蛋白还原酶1(TrxR1)。TrxR，连同 NADPH 和硫氧还蛋白，是硫氧还蛋白系统的一个组成部分，通过二硫键还原酶活性，调节蛋白质二硫醇/二硫键平衡，创建一个关键的抗氧化系统来抵御氧化应激。由于 EECP 也增加了磷酸化 Erk 的表达和 Nrf2的核转位，研究人员认为蜂胶可能通过 Erk 激酶/Nrf2信号通路调节 HO-1、 TrxR1和 GCLM 的表达。
2.8. Propolis as a Cosmetics Additive
Propolis was also studied with regard to its potential application in cosmetics. The research revealed that it can act as a sunscreen agent  and could be used as an ingredient of sunscreen cosmetics . Gismondi et al.  studied its usage as an agent-protecting essential oil, added to sunscreens with the aim of preventing cytotoxic and proradical effects of their components, against damage caused by UV radiation. Lavandula angustifolia Miller essential oil samples, pure or added with 30% ethanol propolis solution at a dose of 1%, were subjected to UV radiation. UV exposure depleted the antioxidant activity of essential oil (DPPH, ABTS, and FRAP assay). Propolis supplementation not only prevented this effect but also considerably increased this parameter in both exposed and nonexposed samples. Those promising results were confirmed by experiment performed on highly metastatic murine B16-F10 melanoma cells. The addition of essential oil samples to culture media caused an increase in cellular GPx, SOD, and CAT activity, but in the case of UV-exposed one, this effect was much less or even slight. However, additional propolis prevented the deterioration of oil properties by UV radiation,as in this case, the results obtained in both oil+propolis+UV and oil+propolis treatments were generally not lower that in the case of essential oil alone.
并对蜂胶在化妆品中的应用前景进行了探讨。研究表明，它可以作为防晒剂，也可以用作防晒化妆品的成分。Gismondi 等人研究了它作为一种保护剂精油的用途，将其添加到防晒霜中，目的是防止其成分的细胞毒性和毒副作用，防止紫外线辐射造成的损害。将纯净的薰衣草精油或加入30% 乙醇蜂胶溶液中，按1% 剂量进行紫外线照射。紫外线照射削弱了精油的抗氧化活性(DPPH，ABTS，和 FRAP 测定)。补充蜂胶不仅防止了这种影响，而且大大增加了这一参数在暴露和非暴露的样本。这些有希望的结果在高转移的小鼠 B16-F10黑色素瘤细胞上得到了证实。在培养基中添加精油样品可以增加细胞内的 GPx、 SOD 和 CAT 活性，但是在紫外线照射的情况下，这种作用要小得多，甚至微乎其微。然而，额外的蜂胶防止了紫外线辐射对油脂性质的损害，在这种情况下，油 + 蜂胶 + 紫外线和油 + 蜂胶处理的结果一般不会低于精油。
3. Bee Pollen
Bee pollen is produced from plant flower pollen, which is collected by bees and mixed with nectar or the salivary gland secretion of the insects. In such form, it is transported, placed on the hind legs, to hives. Then flightless bees mix it with their saliva and pack into honeycombs, covered with a mixture of wax and honey. Under such conditions, the anaerobic fermentation proceeds with the formation of lactic acid, serving as a preservative. The substance, produced in this way, makes a source of nutrients for both adult bees and larvae. The beekeepers collect bee pollen using traps that enable to separate pellets from insects’ legs [71–74].
Pollen food energy is rather high; for instance, bee pollen produced by Apis mellifera, collected in Thailand and containing mainly corn pollen, showed its value to be as much as 397.16 kcal/100 g . In 22 samples of bee pollens collected in Portugal, the obtained values ranged from 396.4 to 411.1 kcal/100 g . It is even named “only perfectly complete food”  or “the life-giving dust” .
3.1. Bee Pollen Composition
The components of bee pollen comprise of a great number of different substances including nutrients (proteins, carbohydrates, and lipids), amino acids (bee pollen is a rich source of leucine, isoleucine, and valine—branched, exogenous amino acids), fatty acids and their esters, vitamins (carotenoids, B, E, H, and folic acid), minerals (macro- and microelements), as well as phenolic organic compounds—flavonoids, phenolic acids, and their derivatives [2, 15, 42, 75, 77–83]. Additionally, the presence of different organic acids (oxalic, tartaric, malic, citric, succinic, acetic, lactic, and gluconic) was found, the latter exhibiting the highest concentration. Among inorganic components macroelements (sodium, potassium, calcium, and magnesium), microelements (iron, zinc, manganese, and copper) as well as some other metals (chromium, aluminium, strontium, tin, nickel, and vanadium) were detected. The content differed considerably in some cases, depending on the source region and plants, particularly as for gluconic acid, potassium, calcium, iron, manganese, and zinc .
蜂花粉的成分包括营养物质(蛋白质、碳水化合物和脂类)、氨基酸(蜂花粉是亮氨酸、异亮氨酸和缬氨酸的丰富来源，外源氨基酸)、脂肪酸及其酯、维生素(类胡萝卜素、 b、 e、 h 和叶酸)、矿物质(宏量元素和微量元素)以及酚类有机化合物(黄酮、酚酸及其衍生物[2,15,42,75,77-83]。不同有机酸(草酸、酒石酸、苹果酸、柠檬酸、琥珀酸、醋酸、乳酸和葡萄糖酸)的含量最高。在无机元素(钠、钾、钙、镁)、微量元素(铁、锌、锰、铜)以及其他金属(铬、铝、锶、锡、镍、钒)中检测到大量元素。在某些情况下，含量差别很大，取决于来源地和植物，特别是葡萄糖酸、钾、钙、铁、锰和锌。
Numerous studies concerning bee pollen included the determination of their detailed composition, particularly considering the biologically active compounds. The following flavonoids and their derivatives were identified as components of rape (Brassica campestris L.) bee pollen: quercetin, naringenin, kaempferol, and isorhamnetin as well as rutin and 3-O-glucosides of quercetin and kaempferol . In the bee pollen from Cistus sp. of Spanish origin, in turn, kaempferol, kaempferol-3-glucoside, quercitin, quercetin-7-rhamnoside, and isorhamnetin were found , whereas in Croatian Cystus incanus L. bee pollen, galangin, kaempferol, chrysin, and pinocembrin were detected . In some samples, the presence of herbacetin, myricetin, tricetin, luteolin, and 3-O-methylquercetin was proved . In Egyptian bee pollen, quercetin, rutin, catechin, epicatechin, kaempferol, apigenin, naringenin, and luteolin were identified . The glucosides of the following anthocyanins, delphinidin, petunidinm, and malvidin were found in bee pollen collected in Spain . According to Silva et al. the analysis of pollen loads collected by stingless bees Melipona rufiventris revealed the presence of different flavonoids, including dihydroquercetin, luteolin, quercetin, isorhamnetin, and isorhamnetin glucoside (isorhamnetin-3-O-(6″-O-E-p-coumaroyl)-β-D-glucopyranoside), which was detected in bee pollen for the first time .
许多关于蜂花粉的研究都包括了其详细成分的测定，特别是考虑到其生物活性化合物。从油菜蜂花粉中分离得到了槲皮素、柚皮素、山奈素、异鼠李素以及槲皮素和3-o- 葡萄糖苷。在产自西班牙的岩蔷薇属植物的蜂花粉中，依次检测到山奈酚、山奈酚 -3- 葡萄糖苷、槲皮素、槲皮素 -7- 鼠李糖苷和异鼠李素，而在克罗地亚的岩蔷薇属植物的花粉中检测到高良姜素、山奈酚、黄素和松叶素。在一些样品中，证实了草药乙素、杨梅素、三甲基黄酮、木犀草素和3-o- 甲基槲皮素的存在。在埃及蜂花粉中分离出槲皮素、芦丁、儿茶素、表儿茶素、山奈素、芹菜素、柚皮素和木犀草素。在西班牙采集的蜂花粉中发现了以下几种花青素的葡萄糖苷: 飞燕草素、小燕草素和麦芽糖苷。根据 Silva 等人对无刺蜜蜂采集的花粉负荷的分析，发现存在不同的黄酮类化合物，包括二氢槲皮素、木犀草素、槲皮素、异鼠李素和异鼠李素葡萄糖苷(异鼠李素 -3-o-(6′-′-o-e-o-p- 香豆素)-β-d- 葡萄糖吡喃葡萄糖苷) ，这是第一次在蜜蜂花粉中检测到。
As for phenolic acids on average, they constitute up to 0.19% of bee pollen and comprise of the derivatives of benzoic, cinnamic, and phenylacetic acids. Gallic acid (3,4,5-trihydroxybenzoic acid) is characterized by a great antioxidant activity .
酚酸平均占蜂花粉的0.19% ，由苯甲酸、肉桂酸和苯乙酸衍生物组成。没食子酸(3,4,5- 三羟基苯甲酸)是一种拥有属性很强的抗氧化剂。
Among the constituents of bee pollen, the following phenolic acids and their esters were reported to be identified: benzoic acid derivatives—p-hydroxybenzoic acid, gallic acid, syringic acid, vanilic acid, and protocatechuic acid—as well as cinnamic acid derivatives—p-coumaric acid, ferulic acid, caffeic acid, and their glycerol esters. Other more complicated derivatives like rosmarinic acid dihexoside as well as amide derivatives of hydroxycinnamic and ferulic acids were also found [14, 41, 71, 74, 77, 86].
3.2. Relationships between the Antioxidant Capacity of Bee Pollen and Its Composition
Bee pollen antioxidant properties have been investigated in many in vitrostudies using DPPH, ABTS+, and FRAP methods. It is well known that the antioxidant capacity of bee pollen is dependent on its content. But the numerous studies, which have been carried out with the aim of determining the composition and properties of different bee pollen samples, proved a considerable diversity of the obtained results. Some studies have shown a strong positive correlation between the total content of phenolic compounds and antioxidant capacity of bee pollen [79, 81], whereas others found no considerable relationships . In the next study , in turn, not phenolic compounds but phenylpropanoid content was found to be correlated with the total antioxidant activity measured by the inhibition of linoleic acid peroxidation. According to Sousa et al. , in turn, flavonols may act as both antioxidants and prooxidants in their reduced and oxidized forms, respectively, whereas anthocyanins act as prooxidants. It has also been found that both content and properties of bee pollen are dependent on the kind of its plant source as well as the conditions of the plants growing like soil or climate. The time of harvesting creates an additional factor affecting these properties [2, 75, 76, 79, 80]. The potential influence of some kind of treatment (freezing as well as freezing and subsequent dehydratation) on the content and properties of bee pollen was also reported. While the chemical composition was not affected by freezing or freezing and subsequent drying, the antioxidant activity was enhanced by freezing and additional drying. The researchers attributed the observed effects to a moisture decrease, leading to antioxidants concentration . The differences in polyphenolic compounds, in total as well as particular types, were also found, showing considerable dispersion, for example, LeBlanc et al.  reported that mimosa bee pollen contained 34.85 mg/g of polyphenolics expressed as gallic acid equivalents, while in the yucca one, only 19.48 mg/g, and in the palm one, 15.91 mg/g were found. The flavonol content in Pyrus communis bee pollen was found to be 1349 mg/100 g, but in the Lamium purpureum one, it reached only 171 g/100 g . In different samples of bee pollen collected in northeastern Brazil in the period of nine months (January–November), flavonoid profiles were found to differ, depending on the time of harvesting and the predominant pollen type .
蜂花粉的抗氧化特性已经在许多体外研究使用 DPPH，ABTS + ，和 FRAP 方法。众所周知，蜂花粉的抗氧化能力取决于其含量。但是为了测定不同蜂花粉样品的组成和性质而进行的大量研究证明了所得结果的相当多样性。一些研究表明，总酚类化合物的含量与蜂花粉的抗氧化能力之间有很强的正相关性，而另一些研究则没有发现相当的关系。在接下来的研究中，反过来，不是酚类化合物而是苯丙素类化合物被发现与通过抑制亚油酸过氧化作用测定的总抗氧化活性相关。根据 Sousa 等人的研究 ，反过来，黄酮醇在其还原和氧化形式中分别作为抗氧化剂和促氧化剂，而花青素作为促氧化剂。研究还发现，蜂花粉的含量和性质取决于其植物来源的种类和土壤、气候等植物生长的条件。收获的时间增加了一个影响这些特性的因素[2,75,76,79,80]。本文还报道了不同处理方法(冷冻、冷冻和后续脱水)对蜂花粉含量和性质的潜在影响。虽然化学成份没有受到冷冻或冷冻和随后的干燥的影响，但是抗氧化活性通过冷冻和额外的干燥得到增强。研究人员将观察到的影响归因于水分减少，导致抗氧化剂浓度。多酚类化合物在总体和特定类型上的差异也被发现，显示出相当大的分散性，例如 LeBlanc 等人的报告说，含羞草蜂花粉含有34.85 mg/g 以没食子酸当量表达的多酚类化合物，而在丝兰花粉中，只有19.48 mg/g，在棕榈花粉中，15.91 mg/g。梨蜂花粉中黄酮醇含量为1349mg/100g，而紫叶梨仅为171g/100g 。在巴西东北部9个月(1月至11月)采集的不同蜂花粉样品中，黄酮类化合物的含量不同，这取决于采集的时间和主要的花粉类型。
Similar to the case of propolis, the research has shown that the type of the extraction solvent used may affect the properties of the pollen extract to a considerable degree (Table 1). This is connected with different solubilities of particular components of bee pollen in solvents of diverse polarities. It was proved that the application of nonpolar solvents resulted in extracts of very low antioxidant activity, whereas the polar ones allowed obtaining the better antioxidant properties. However, considerable differences were observed even in the case of the application of solvents of similar polarity [11, 41, 42]. The optimal condition for bee pollen extraction was studied by Kim et al. . In their experiment, the total extract obtained by extraction with 80% methanol (twice) was then successively partitioned using solvents of different polarities: n-hexane, dichloromethane, ethyl acetate, and n-butanol. Ethyl acetate and n-butanol fractions exhibited the greatest activity, so in the next step, the optimal conditions of extraction were estimated by response surface methodology using the Box-Behnken design (BBD) with three-level three-factor. The variables were ethyl acetate concentration in methanol, temperature, and time. The solvent concentration proved to exhibit the greatest impact, and the optimal parameters were established as follows: 69.6% ethyl acetate in methanol, 10.0°C and 24.2 h. The calculated values were confirmed experimentally, as the extract obtained under the theoretically estimated conditions showed the antioxidant activity and tyrosinase inhibition very similar to those predicted by statistical methods . The dependence of extract properties on the used extraction solvent was also confirmed in the animal research. In rats with induced hind paw edema, the oral administration of bee pollen bulk showed mild suppressing properties, and water extract had practically no effect, while ethanol extract displayed the greatest effectiveness .
类似的情况下，蜂胶，研究表明，提取溶剂的类型可能会影响花粉提取物的性质相当大的程度(表1)。这与蜂花粉特定组分在不同极性溶剂中的不同溶解度有关。结果表明，非极性溶剂的应用使得提取物的抗氧化活性非常低，而极性溶剂的应用使得提取物具有较好的抗氧化性能。然而，即使在应用相似极性的溶剂[11,41,42]的情况下，也观察到相当大的差别。研究了蜂花粉提取的最佳条件。本实验采用80% 甲醇(2次)萃取法得到的总提取物，然后用正己烷、二氯甲烷、乙酸乙酯、正丁醇等不同极性的溶剂依次进行分离。乙酸乙酯和正丁醇萃取部分的活性最高，因此在下一步中，采用三因素三水平 Box-Behnken 设计(BBD)对反应曲面法的最佳提取条件进行了优化。考察了乙酸乙酯在甲醇中的浓度、反应温度和反应时间对反应的影响。实验结果表明，溶剂浓度的影响最大，确定了最佳工艺参数为: 69.6% 乙酸乙酯在甲醇中，10.0 ° c，24.2 h。实验证实了计算值，因为在理论估计条件下获得的提取物显示了抗氧化活性和酪氨酸酶抑制非常相似的预测统计方法。动物实验也证实了萃取物性质对萃取溶剂用量的依赖性。在大鼠足肿胀模型中，蜂花粉块的口服给药表现出轻微的抑制作用，水提取物几乎没有作用，而乙醇提取物表现出最大的效果。
3.3. Bee Pollen Role in Mitigation of Chemotherapy Side Effect
Bee pollen has been found both to alleviate the deterioration of antioxidant barrier and instead of nad to inhibit lipid peroxidation process following chemotherapy.
研究发现蜂花粉既可以减轻抗氧化屏障的恶化，又可以代替 nad 抑制化疗后的脂质过氧化过程。
Huang et al.  found that the intraperitoneal treatment with cisplatin resulted in the extensive deterioration of liver and kidney functions. This harmful effect involved a significant increase in the concentration of a lipid peroxidation marker MDA and iNOS as well as a well-marked depletion of activities of chosen antioxidant enzymes. However, the additional intragastrical pretreatment with Schisandra chinensis bee pollen was able to alleviate these effects in a dose-dependent way. The additional evidence for the antioxidant influence of bee pollen was the comparison of the described results with the effects observed in animals treated according to the same design with cisplatin and an acknowledged antioxidant vitamin—ascorbic acid—whose impact was comparable with that observed for bee pollen (Table 3). The relationships between antioxidant properties and protective influence of bee pollen against cisplatin were also confirmed by Tohamy et al. . In a study performed on cisplatin-exposed male mice, distinct symptoms of oxidative stress in organs (kidney, liver, and testis) were observed as the lipid peroxidation process was greatly intensified, while CAT activity and GSH concentration markedly depressed. However, the oral coadministration of Egyptian bee pollen water extract significantly alleviated prooxidant changes. Moreover, bee pollen alone decreased neither CAT nor GSH and distinctly inhibited lipid peroxidation in the kidney and testes. As the studied organs show a great vulnerability to the toxic action of cisplatin, the obtained results made the authors suggest the potential effectiveness of bee pollen at alleviating cisplatin-induced side effects. It seems to be worth noticing that in this case, the bee pollen proved to be effective even when administered after cisplatin application. The experiment performed by scientists from Malaysia supported the abovementioned findings. According to the authors, the methanol extract of bee pollen of Malaysian stingless bee (Lepidotrigona terminata (L. terminata)), displaying a distinct, dose-dependent antioxidant activity, was also effective for the antiproliferation of cells. Such an effect was observed in the case of both cancer (MCF-7) and normal (L929) ones with the IC50 value being much lower for cancer cells. Then the effect of cisplatin alone and in cotreatment with bee pollen extract was studied on MCF-7 cells. Cell proliferation was distinctly inhibited by cisplatin, and additional bee pollen revealed a potentializing influence on cisplatin action. In the next stage, the analysis of the influence of bee pollen and cisplatin combination on MCF-7 cell line was performed using CompuSyn software to evaluate if these agents work in an antagonistic, synergic, or additive way. The statistical analysis allowed the authors to suggest that these two substances acted in synergistic way. In conclusion, the researchers suggested the possibility of the application of the studied bee pollen, capable of aiming at potentiating the effectiveness of the therapy and allowing the decrease of the dose of chemoprotective drugs  (Table 3).
黄等人发现，腹腔内的治疗与顺铂导致广泛恶化的肝脏和肾功能。这种有害影响包括脂质过氧化标记物 MDA 和 iNOS 浓度的显著增加以及所选择的抗氧化酶活性的显著衰竭。然而，额外的胃内预处理五味子蜂花粉能够以剂量依赖的方式减轻这些影响。蜂花粉抗氧化作用的另一个证据是上述结果与按顺铂和公认的抗氧化剂维生素抗坏血酸相同设计处理的动物中观察到的效果的比较，后者的影响与蜂花粉中观察到的效果相当(表3)。蜂花粉的抗氧化特性与其对顺铂的保护作用之间的关系也得到了 Tohamy 等人的证实。在一项对顺铂暴露的雄性小鼠进行的研究中，观察到器官(肾脏、肝脏和睾丸)有明显的氧化应激症状，因为脂质过氧化过程大大加强，而 CAT 活性和 GSH 浓度明显降低。然而，口服埃及蜂花粉水提取物可显著减轻促氧化剂的变化。此外，单独使用蜂花粉既不降低 CAT 也不降低 GSH 含量，并且明显抑制肾脏和睾丸中的脂质过氧化。由于所研究的器官对顺铂的毒性作用极为敏感，所得结果提示蜂花粉对缓解顺铂所致的副作用具有潜在的有效性。值得注意的是，在这种情况下，蜂花粉被证明是有效的，即使在顺铂应用后。来自马来西亚的科学家进行的实验支持了上述发现。作者认为，马来西亚无刺蜜蜂花粉甲醇提取物具有明显的剂量依赖性抗氧化活性，对细胞增殖也有抑制作用。这种效应在癌症(MCF-7)和正常(L929)的病例中都可以观察到，癌细胞的 IC50值要低得多。然后观察顺铂单独及与蜂花粉提取物联合处理对 MCF-7细胞的影响。顺铂对细胞增殖有明显的抑制作用，蜂花粉对顺铂作用有潜在的抑制作用。接下来，利用 CompuSyn 软件分析蜂花粉和顺铂联合作用对 MCF-7细胞系的影响，评价这些药物是否以拮抗、协同或加和的方式发挥作用。统计分析允许作者提出，这两种物质相互作用的方式。总之，研究人员建议应用研究的蜂花粉的可能性，能够增强治疗的有效性，并允许减少化学保护药物的剂量(表3)。
|Source 资料来源Toxic factor毒性因素Harmful effects of a toxic factor 毒性因素的有害影响The type of bee pollen and the way of application 蜂花粉的种类及应用方法Effects of bee pollen coadministration 蜂花粉共管效应的研究Mitigation effect of bee pollen on chemiotherapeutic agents 蜂花粉对化疗药物的缓解作用 Huang et al. [黄等Cisplatin-induced toxicity in rats, (8 mg/kg b.w.目的: 探讨顺铂对大鼠的毒性作用i.p.in single dose) on the 7th day of the 12-day-experiment 12天实验的第7天↑ MDA and iNOS: liver and kidney MDA 与 iNOS: 肝、肾|
↓ SOD, CAT, and GSH: liver and kidney 和 GSH: 肝脏和肾脏Schisandra chinensis 五味子 bee pollen extracted with 70% ethanol, 400, 800, and 1200 mg/kg b.w.蜂花粉用70% 乙醇、400、800和1200毫克/千克 b.w. 提取p.o., 假释官, 12 days12天↓ MDA in liver 4. 肝脏里的 MDA∗∗ 第 x 次/次 and kidney 还有肾脏
↓ iNOS in liver and kidney 4. 肝脏和肾脏的 iNOS∗∗ 第 x 次/次
↑ SOD in liver 肝脏中∗∗ 第 x 次/次 and kidney 还有肾脏∗∗ 第 x 次/次
↑ CAT in the liver and kidney 肝肾中的 CAT ↑
↑ GSH in the liver and kidney 肝、肾中谷胱甘肽↑Tohamy et al. [ 托哈米等人Cisplatin-induced toxicity in male mice (2.8 mg/kg b.w. 顺铂对雄性小鼠的毒性作用(2.8 mg/kg b.w.)i.p. twice/week for 3 weeks) 。每星期两次，连续三星期)↑ Lipid peroxidation in liver, kidney and testis 肝、肾和睾丸中↑脂质过氧化
↓ CAT and GSH in the liver, kidney, and testis 4、肝脏、肾脏和睾丸中的过氧化氢酶和谷胱甘肽Water, Egyptian bee pollen extract, 140 mg/kg b.w. once a day orally, during the last 2 weeks of cisplatin exposure 水，埃及蜂花粉提取物，140毫克/公斤每天一次口服，在顺铂暴露的最后2周↓ Lipid peroxidation in the kidney, liver, and testis 4、肾脏、肝脏和睾丸的脂质过氧化
↑ CAT and GSH in the kidney, liver, and testis 肾、肝和睾丸中 CAT ↑和 GSH 的表达Mitigation effect of bee pollen on other toxic agents 蜂花粉对其他有毒物质的缓解作用 Ferreira et al. [ 费雷拉等人Tebuconazole-exposed fish (catfish jundiá), 0.88 mg/L (16.6% of 96 h LC 戊唑醇暴露鱼(鲶鱼俊迪亚) ，0.88 mg/L (96 h LC 的16.6%)50)
96 hours 96小时↑ Lipid peroxidation in the liver, kidney, and brain 会影响肝脏、肾脏和大脑中的脂质过氧化
↓ SOD in liver 4. 肝脏里的 SOD
↑ CAT in the liver and brain 脑和肝中的 CAT ↑Bee pollen; 0.01, 0.03, and 0.05 g/L, environmental exposure 蜂花粉; 0.01,0.03,0.05 g/L，环境暴露↓ Lipid peroxidation in the liver, kidney 4. 肝脏、肾脏的脂质过氧化∗∗ 第 x 次/次, and brain ，以及大脑∗∗ 第 x 次/次
↑ SOD in the liver肝脏中的 SOD∗∗ 第 x 次/次
↓ CAT in liver 在肝脏的↓ CAT∗∗ 第 x 次/次: low and high doses 低剂量和高剂量
↓ CAT in brain 在脑海中的↓ CAT∗∗ 第 x 次/次Yıldız et al. [等人Carbon tetrachloride-induced hepatotoxicity in rats (0.85 mL/kg b. w. 四氯化碳致大鼠肝毒性(0.85 mL/kg · b · wi.p., 7 days) . ，7天)↑ plasma ALT and AST 血浆 ALT、 AST
↑ MDA in liver, RBC and plasma; 肝、红细胞和血浆中 MDA ↑ ;
↓ SOD in plasma, RBC and liver 4. 血浆、红细胞和肝脏中的 SODBee pollen collected during flowering season in Turkey (Western Black Sea region) with dominant component chestnut 在土耳其(黑海西部地区)的花期采集了以板栗为主要成分的蜂花粉sativa 大麻纤维卷 pollen (>45%), 200 mg/kg/day orally, 400 mg/kg/day orally, 7 days 花粉(> 45%) ，口服200mg/kg/d，口服400mg/kg/d，7d↓ Plasma ALT: high dose 4、海洛因: 海洛因
↓ Plasma AST 4. Plasma ↓ AST
↓ MDA in the plasma, RBC, and liver 4. 血浆中的 MDA，红细胞和肝脏Almaraz-Abarca et al. Bromobenzene-induced hepatotoxicity in mice, 94.211 溴苯致小鼠肝毒性，94.211μg/mL in oil, 200 每毫升油，200克μL orally L 口服↑ Lipid peroxidation liver 脂质过氧化Bee pollen from mesquite ( 豆科牧豆科植物的蜂花粉Prosopis juliflora 牧豆树) collected in April in Mexico, extracts of two flavonol concentration (9.794 ) ，其中两种黄酮醇的提取物(9.794μg/mL and 21.751 克/毫升及21.751克/毫升μg/mL), 200 克/毫升) ，200毫升μL orally L 口服↓ Liver lipid peroxidation—only the higher dose 4. 肝脏的脂质过氧化作用ーー只是剂量较高Ketkar [ 凯特卡Chronic exercise-induced oxidative stress in rats, 4 weeks 慢性运动诱导的大鼠氧化应激，4周↓ Gastrocnemius muscle SOD and GSH 4. 腓肠肌的 SOD 和 GSH
↑ Gastrocnemius muscle MDA and NO 腓肠肌和 NO
↓ Weight of gastrocnemius muscle and body 4. 腓肠肌和身体的重量The neat and processed (1 mg of bee pollen : 500 mg of Captex 355 : 750 mg of Tween 80) monofloral Indian mustard bee pollen, 100, 200, or 300 mg/kg daily, orally 单花印度芥菜蜂花粉，每日100、200或300 mg/kg↑ SOD and GSH in gastrocnemius muscle 腓肠肌中的 SOD 和 GSH
↓ MDA in gastrocnemius muscle: neat 腓肠肌的↓ MDA: 干净利落∗∗ 第 x 次/次: high dose, processed: all doses 高剂量，加工过的，全部剂量
↓ NO in gastrocnemius muscle neat 公司的名字↓在腓肠肌∗∗ 第 x 次/次: higher ones, processed: all doses 高剂量的，加工过的，全部剂量
↑ Body weight 体重
↑ Gastrocnemius muscle weight neat 腓肠肌重量∗∗ 第 x 次/次: high one, processed 高一，加工完毕∗∗ 第 x 次/次: higher doses : 更高剂量
The positive effects increase along with the increase in the dose 随着剂量的增加，正效应增加
|ALT: alanine aminotransferase; AST: aspartate aminotransferase; CAT: catalase; GSH: reduced glutathione, iNOS: inducible nitric oxide synthase; MDA: malondialdehyde; NO: nitrogen oxide; RBC: red blood cell; SOD: superoxide dismutase; ↓: decrease; ↑: increase; 谷丙转氨酶: 丙氨酸氨基转移酶; 天冬氨酸氨基转移酶; CAT: 过氧化氢酶; GSH: 谷胱甘肽，iNOS: 诱导型一氧化氮合酶; MDA: 丙二醛; NO: 氮氧化物; 红细胞: 红细胞; SOD: 超氧化物歧化酶; ↑ : 减少; ↑ : 增加; ↑ :∗∗ 第 x 次/次the effect depended on used dose. 效果取决于使用剂量|
Table 3 表三The protective effects of bee pollen against prooxidant action of different harmful factors. 蜂花粉对不同有害因子促氧化作用的保护作用
3.4. Bee Pollen as a Protective Agent against Prooxidants’ Toxicity
Antioxidant properties of bee pollen encouraged the research concerning its application as an agent preventing or alleviating harmful oxidative processes occurring in organisms or caused by different factors. The performed studies included interesting, diverse issues and resulted in considerably promising findings.
Turkish scientists investigated bee pollen as a protective agent against carbon tetrachloride hepatotoxicity. The results were additionally compared with those obtained for silibinin, an active component of silymarin (a plant-origin substance used in hepatic disorder cure). All the applied treatments caused a decrease in body weight gain. However, this effect was the least in the case of the coadministration of CCl4 and the higher dose of bee pollen. A significant enhancement of liver injury markers—plasma activity of transferases ALT and AST observed in CCl4-exposed rats—was considerably alleviated in animals cotreated with both silibinin and bee pollen. Furthermore, the effect of the higher bee pollen dose was not markedly different from that observed in the case of silibinin. Liver and plasma MDA were found to be significantly increased by CCl4 exposure, and again, both studied protective agents were capable of reversing this effect, although that in liver silibinin was more effective. The SOD activity in plasma, RBC, and liver was depressed in CCl4-exposed rats, and in this case, both silibinin and bee pollen proved to lack protective influence. As chestnut bee pollen was found to contain antioxidants and a substantial antioxidant capacity measured by FRAP and DPPH methods, the authors suggested the possibility of replacing silibinin by bee pollen in liver disorders cure, all the more because the harmful effects were observed in silibinin-treated animals—decrease in body weight gain, severe diarrhea, and, consequently, mortality  (Table 3).
土耳其科学家研究了蜂花粉作为一种保护剂对四氯化碳肝毒性。这些结果还与水飞蓟宾的结果进行了比较，水飞蓟宾是水飞蓟素(一种用于肝病治疗的植物来源物质)的活性成分。所有应用的处理都导致体重增加减少。但是，在四氯化碳和蜂花粉高剂量的共同作用下，这种作用最小。水飞蓟宾和蜂花粉联合处理能显著减轻 ccl_4暴露大鼠肝损伤标志物——血浆转移酶 ALT 和 AST 活性的升高。此外，较高的蜂花粉剂量的影响并没有显着不同的情况下观察水飞蓟宾。接触四氯化碳可显著增加肝脏和血浆中的丙二醛含量，同样，研究中的两种保护剂都能够逆转这种作用，尽管水飞蓟宾对肝脏的保护作用更为有效。四氯化碳染毒大鼠血浆、红细胞和肝脏中 SOD 活性降低，水飞蓟宾和蜂花粉对肝脏中 SOD 活性均无保护作用。由于栗子蜂花粉含有抗氧化剂，并通过 FRAP 和 DPPH 法测定其抗氧化能力，作者认为用蜂花粉替代水飞蓟宾治疗肝病是可行的，特别是水飞蓟宾治疗动物的不良反应明显，体重下降，严重腹泻，死亡率降低(表3)。
Bee pollen was also shown to alleviate aflatoxin–induced oxidative processes in spleen by a decrease in the H2O2 level accompanied by GSH enhancement and NO proper generation .
蜂花粉也被证明可以减轻黄曲霉毒素引起的脾脏氧化过程，降低 H2O2水平，同时还可以增强谷胱甘肽和 NO 的适当生成。
In another animal study, bee pollen was investigated with regard to its possible application as an agent alleviating stress induced by exercise. Taking into account the fact that nutrients contained in bee pollen are barely absorbed in the gastrointestinal track due to the tough coat enclosing the components inside and making digestion difficult to proceed, the authors carried out a very interesting comparison of the neat and processed monofloral Indian mustard bee pollen. The processed one was obtained by mixing with an edible lipid-surfactant mixture (Captex 355 and Tween 80 in different ratios). The composition of the lipid-surfactant mixture influenced the total polyphenol content in the obtained processed samples, with the ratio of 1 mg of bee pollen : 500 mg of Captex 355 : 750 mg of Tween 80 showing the highest value. For this reason, that sample was chosen for further studying using an animal model as a protective agent against oxidative changes caused by exercise. In rats subjected to chronic exercise, the distinct evidence of oxidative stress was shown as SOD, and GSH were found to be markedly decreased, while MDA and NO significantly increased in the gastrocnemius muscle. Additionally, exercised animals displayed significantly decreased body and gastrocnemius muscle weights compared to the control. All these exercise-induced changes were partially reversed by oral treatment with both neat and processed bee pollen in a dose-dependent way, and the processed one proved to have much greater efficiency. Furthermore, the processed bee pollen, given alone to nonexercised animals, generally did not substantially affect the studied parameters in comparison with the control. The authors concluded that processing improved the availability of bee pollen nutrients and subsequently all beneficial effects  (Table 3).
在另一项动物研究中，研究了蜂花粉作为运动应激缓解剂的可能应用。考虑到蜂花粉中含有的营养物质在胃肠道中几乎不被吸收，这是由于蜂花粉中的成分被坚硬的外壳包裹，使得消化难以进行，作者对纯的和经过加工的单花印度芥菜蜂花粉进行了一次非常有趣的比较。将可食性脂质-表面活性剂混合物(Captex 355和 Tween 80按不同比例混合)加工得到。脂质-表面活性剂混合物的组成对处理后的样品中总多酚含量有影响，蜂花粉1mg: Captex 355500mg: Tween 80750mg 的比例最大。基于这个原因，选择这个样本进行进一步的研究，使用动物模型作为保护剂，对抗运动引起的氧化变化。在长期运动的大鼠中，氧化应激的明显证据表现为 SOD，GSH 明显降低，而 MDA 和 NO 在腓肠肌中明显升高。此外，与对照组相比，运动过的动物体重和腓肠肌显著下降。所有这些运动诱导的变化都通过口服纯蜂花粉和加工蜂花粉治疗得到部分逆转，并且呈剂量依赖性，加工蜂花粉治疗的效果更好。此外，处理过的蜂花粉，单独给予未运动的动物，一般没有实质上影响研究的参数相比，对照。作者得出结论，加工提高了蜂花粉营养物质的利用率，随后产生了所有有益的效果(表3)。
Bee pollen has also been found to show some protective effect against oxidative damage observed in fish environmentally exposed to tebuconazole (a fungicide of high toxicity to aquatic organisms), but the obtained results were not unambiguous. Tebuconazole caused a considerable intensification of lipid peroxidation in chosen organs and a decrease in liver SOD. Additional bee pollen considerably reversed these effects. However, in the case of CAT, the obtained results were not so promising. Tebuconazole alone enhanced its activity in the liver and brain and showed no significant effect in kidney. The cotreatment with bee pollen displayed diverse effects, depending on dose and organ, with no determined tendency. But the most important fact was a significant CAT decrease versus control observed in the kidney and brain of fish exposed to higher concentrations of bee pollen. Furthermore, bee pollen alone also depressed CAT in those organs compared to control .
研究还发现，蜂花粉对环境暴露于戊唑醇(一种对水生生物具有高毒性的杀菌剂)的鱼体内的氧化损伤有一定的保护作用，但所得结果并不明确。戊唑醇引起选定器官中脂质过氧化的相当强化和肝 SOD 的减少。额外的蜂花粉相当程度上逆转了这些影响。然而，对于计算机辅助翻译，所得到的结果并不那么令人满意。戊唑醇单独作用于肝脏和脑组织，对肾脏无明显影响。蜂花粉辅助处理的效果因剂量和器官的不同而有所差异，但无明显变化趋势。但最重要的事实是，与对照组相比，暴露在高浓度蜂花粉环境中的鱼类，其肾脏和大脑中的 CAT 明显减少。此外，与对照组相比，单独使用蜂花粉也会抑制这些器官中的 CAT。
Almaraz-Abarca et al.  investigated the properties of bee pollen from mesquite (Prosopis juliflora) collected in Mexico and also received inconsistent results. Bee pollen extracts of two flavonol concentrations prevented lipid peroxidation observed in the liver of mice exposed to bromobenzene but the results reached statistical significance only in the case of higher dose. However, the extract of higher concentration given alone caused a significant intensification of liver lipid peroxidation in mouse, comparably with the one observed in bromobenzene-treated animals. Interestingly, the extract of lower concentration showed a great antioxidant effect as lipid peroxidation in this case was even decreased when compared to the control with no treatment. The authors concluded that in the absence of any oxidative stress-inducing factor, the administration of the high concentration of flavonols itself may induce oxidative damage occurrence. According to the authors, the confirmation of such an assumption could be the reports revealing that both polyphenols and an acknowledged antioxidant vitamin C may act as prooxidants in the presence of transient metal ions. Despite the similarity of the effects, the authors do not postulate the similarity of the mechanism of prooxidant influence of flavonoids and vitamin C. However, these outcomes show the necessity of proper precaution in the application of bee pollen, particularly in considering the dose (Table 3).
Almaraz-Abarca 等人调查了在墨西哥采集的牧豆树(mesquite)蜂花粉的特性，也得到了不一致的结果。蜂花粉提取物的2种黄酮醇浓度抑制了溴苯暴露小鼠肝脏中观察到的脂质过氧化，但结果只有在较高剂量的情况下才达到统计学意义。然而，单独给予较高浓度的提取物会导致小鼠肝脏脂质过氧化明显增强，与溴苯处理的动物相比。有趣的是，较低浓度的提取物表现出很强的抗氧化作用，因为在这种情况下，与未经治疗的对照组相比，脂质过氧化甚至减少了。作者得出结论，在没有任何氧化应激诱导因素的情况下，高浓度的黄酮醇本身可能诱导氧化损伤的发生。根据作者的说法，这一假设的证实可能是报告揭示了多酚和公认的抗氧化剂维生素 c 可能在短暂的金属离子存在时起促氧化剂的作用。尽管效果相似，作者并没有假定黄酮类化合物和维生素 c 促氧化作用的机制相似，但是，这些结果表明在使用蜂花粉时，特别是在考虑剂量时，有必要采取适当的预防措施(表3)。
Similar conclusions were drawn from the experiment performed by Sousa et al. . Studying bee pollen from Echium plantagineum L., they used three different substances: its extract enriched in flavonols (fraction I), its extract enriched in anthocyanins (fraction II), and the combination of both extracts (the whole extract). Both I and II extracts contained kaempferol glucosides, and additionally in fraction II anthocyanins, glucosides of delphinidin, petunidin, and malvadin were detected. All three extracts were studied in vitro considering their influence on viability, reactive species, and antioxidants in Caco-2 cells. Interestingly, the extracts exhibited varied activity under different conditions. As for cells not subjected to any factor, neither fraction I nor fraction II caused any changes in cell viability measured by MTT assay, while the whole extract containing the combination of flavonols and anthocyanins, used in high concentration (20 mg/mL), caused a significant cellular viability depression. The effect of pretreatment with bee pollen extracts on the viability of cells induced by butyl hydroperoxide (t-BHP), whose presence stimulated reactive oxygen species production, was also evaluated. Herein, fraction II used in higher concentrations (2.5–20 mg/mL) and the whole extract (2.5–10 mg/mL) intensified t-BHP-induced harmful effect, while the highest concentrations of fraction I showed some insignificant but distinct protective effect. Additionally, the percentage values of cell viability, obtained for the whole extract, applied in concentration of 20 mg/mL, were the same, regardless of t-BHP presence or absence. These outcomes allowed the authors to suggest that anthocyanins acted as prooxidants, while flavonols supported antioxidant barrier but, in their oxidized forms, might also contribute to prooxidant processes. Lower concentrations of fractions I and II were also efficient at decreasing reactive species level. Interestingly, high concentration (20 mg/mL) prevented reactive species production only after a short t-BHP exposure. Along with the exposure, lengthening fraction II was proved to be ineffective, while fraction I began to act as a prooxidant as reactive species generation was shown to be enhanced. The whole extract did not show a significant antioxidant efficacy, and the higher concentrations exerted even prooxidant effect, particularly after a longer time of t-BHP exposure. Next, reduced glutathione was determined in the cells treated with different concentrations of the studied extracts, with and without subsequent exposure to t-BHP. The only effects were observed in cells exposed to t-BHP—a significant GSH increase after pretreatment with 20 mg/mL of the whole extract and a slight enhancement after using higher doses of fraction I. The authors tried to explain those complex observations pointing to GSH participation in t-BHP defusing, the bioactivation of t-BHP by cytochrome P450 as well as the inhibition of the influence of flavonoids (mainly anthocyanins) on cytochrome P450 enzymatic activity, the capacity of anthocyanins for being transformed into radicals, as well as the ability of t-BHP to form species more active than those produced by the process of its biotransformation. Concluding, the authors underlined the necessity of taking the proper precautions in using bee pollen, particularly considering the dose.
从 Sousa 等人的实验中也得出了类似的结论。研究车前叶蓝蓟的蜂花粉时，他们使用了三种不同的物质: 富含黄酮醇的提取物(分数 i) ，富含花青素的提取物(分数 II) ，以及两种提取物的组合(整个提取物)。在 i 和 II 提取物中均含有山奈酚葡萄糖苷，此外还检测到分数 II 中的花青素、飞燕草苷、小豆豆苷和马尔瓦丁的葡萄糖苷。考虑到三种提取物对 Caco-2细胞活力、活性物质和抗氧化剂的影响，在体外进行了研究。有趣的是，提取物在不同条件下表现出不同的活性。对于没有受到任何因子影响的细胞，MTT 法测定的分数 i 和分数 II 都没有引起细胞活力的变化，而含有黄酮醇和花青素的高浓度(20mg/ml)的整个提取物引起了显著的细胞活力下降。研究了蜂花粉提取物预处理对过氧化氢(t-BHP)诱导的细胞活力的影响，t-BHP 的存在促进了活性氧类的产生。高浓度(2.5ー20mg/ml)和全提取物(2.5ー10mg/ml)处理组分 II 对 t-bhp- 有明显的保护作用，高浓度组分 II 对 t-bhp- 有明显的保护作用。此外，不论 t-BHP 是否存在，在20mg/ml 浓度下，全部提取物的细胞活力百分比值都是相同的。这些结果使作者认为花青素起到了促氧化剂的作用，而黄酮醇支持抗氧化屏障，但是，在它们的氧化形式中，也可能有助于促氧化过程。低浓度的组分 i 和 II 对降低反应物种水平也有效。有趣的是，高浓度(20毫克/毫升)仅仅在短时间的 t-BHP 接触后就阻止了活性物质的产生。随着暴露时间的延长，延长部分 II 被证明是无效的，而部分 i 开始作为一种促氧化剂，因为活性物种的产生被证明是增强的。整个提取物没有显示出明显的抗氧化功效，更高的浓度甚至产生了促氧化作用，特别是在较长时间的 t-BHP 暴露之后。接下来，用不同浓度的研究提取物处理细胞，测定细胞中的谷胱甘肽含量，以及随后是否接触 t-BHP。在 t-bhp 染毒的细胞中观察到了唯一的影响ーー预处理20mg/ml 提取物后 GSH 明显增加，高剂量 i 染毒后 GSH 略有增加。作者试图解释这些复杂的观察结果，指出谷胱甘肽参与 t-BHP 化解，细胞色素 P450对 t-BHP 的生物活化，黄酮类化合物(主要是黄酮类化合物)对细胞色素 P450酶活性的影响，花青素转化为自由基的能力，以及 t-BHP 形成物种的能力比其生物转化作用过程中产生的更活跃。最后，作者强调了在使用蜂花粉时采取适当预防措施的必要性，特别是考虑到剂量。
3.5. Bee Pollen in Cosmetics
The beneficial influence of bee pollen on skin condition has been known from ancient times. Recently, as the return to natural medicine agents is being observed, some studies have been carried out with the aim of clarifying the mechanism of these effects.
Sun et al. , in their experimental study, confirmed the possibility of bee pollen using in skin therapy, connected with its antioxidant ability. Chinese scientists studied composition, antioxidant activity, as well as its influence on the melanogenesis of two rape bee pollen extracts containing free or bound phenolics. Both extracts exhibited antioxidant capacity measured by DPPH, ABTS, and FRAP methods, although the free phenolic one proved to be much more effective as an antioxidant. In the next stage of the study, the effect on melanin synthesis was investigated. Tyrosinase activity was inhibited in a dose-dependent way by both extracts with the free one being more effective. Since some phenolic compounds also showed the same effect, the authors assumed the existence of some relationship between phenolic profile and inhibitory influence on tyrosinase activity. The tyrosinase inhibition was studied considering the fact that melatonin, despite its protective role for the skin, in excessive amounts, may itself exert harmful effects like reactive oxygen species generation and pigmentation. The effect of free phenolic extract of bee pollen on melanogenesis was also studied using B16 mouse melanoma cells. The studied substance decreased intracellular tyrosinase activity and melanin relative content in a very distinct, dose-dependent way. Moreover, taking into account the connection between the melanogenesis and generation of oxygen reactive species and consequently the role of intracellular reducing activity in melanogenesis regulation, GSH/GSSG value was measured. Free phenolic bee pollen extract again showed a high effectiveness at increasing reducing power by enhancing intracellular GSH/GSSG ratio, indirectly contributing to the depression of melanin synthesis. All the presented results point to the usefulness of bee pollen in protecting the cell against abnormal melanogenesis, which cannot be overrated as melanin is responsible for numerous skin disorders, from freckles up to malignant melanoma . The usefulness of bee pollen in manufacturing cosmetics protecting the skin against hyperpigmentation and oxidative stress was also confirmed by Korean scientists .
在他们的实验研究中，证实了蜂花粉用于皮肤治疗的可能性，这与蜂花粉的抗氧化能力有关。中国科学家研究了含有自由或结合酚的两种油菜蜂花粉提取物的成分、抗氧化活性及其对黑素形成的影响。这两种提取物都表现出抗氧化能力测定 DPPH，ABTS，和 FRAP 方法，虽然游离酚一被证明是更有效的抗氧化剂。在下一阶段的研究，对黑色素合成的影响进行了调查。两种提取物均有剂量依赖性地抑制酪氨酸酶活性，游离提取物对酪氨酸酶活性的抑制作用更强。由于一些酚类化合物也表现出同样的作用，作者认为酚类化合物的组成与抑制酪氨酸酶活性的影响有一定的关系。考虑到褪黑激素虽然对皮肤有保护作用，但过量的褪黑激素本身可能会对皮肤产生有害的影响，如活性氧类和色素沉着，所以研究了酪氨酸酶抑制。用 B16小鼠黑色素瘤细胞研究了蜂花粉中游离酚类提取物对黑素合成的影响。所研究的物质降低细胞内酪氨酸酶活性和黑色素相对含量的非常明显，剂量依赖的方式。此外，考虑到黑素生成和氧活性物质的产生之间的联系以及细胞内还原活性在黑素生成调节中的作用，测定了 GSH/GSSG 值。游离酚类蜂花粉提取物通过提高细胞内 GSH/GSSG 比值，再次显示出较高的还原能力，间接导致黑色素合成的抑制。所有提交的结果都指出蜂花粉在保护细胞免受异常黑素生成方面的作用，不能过高估计黑色素，因为黑色素是许多皮肤疾病的罪魁祸首，从雀斑到黑色素瘤。韩国科学家也证实了蜂花粉在制造化妆品中的用途，这种化妆品可以保护皮肤免受色素沉着和氧化应激的伤害。
4. Royal Jelly
Royal jelly is a secretion from mandibular and hypopharyngeal glands of young bees of the Apis mellifera species . This is a white or yellowish cream substance that makes food for young bee larvae (but no longer than three days, and then they are fed with a mixture of pollen, nectar, and honey) and the only food for the queen in both the larval and adult stages [95–97]. This difference in the way of feeding is considered as the main factor responsible for the differentiation in the development of bee workers and the queen. In comparison to the workers’ food, the royal jelly contains less water and four times more sugars, more proteins, and different concentrations of some mineral salts . This unique composition of royal jelly leads to changes in gene expression (most probably through epigenetic mechanisms) which allows, for instance, full ovarian development to proceed . Thanks to royal jelly, the queen could live up to five years (workers usually live about 45 days) and lay about 2000–3000 eggs a day . For commercial use, royal jelly is collected from the queen’s cells, as they are the richest sources of this product—it is produced in a much larger amount than queen larvae are able to consume . According to some sources, the annual production of royal jelly amounts to several thousand tons—about 2000 tons are produced only in China .
蜂王浆是意大利蜜蜂幼蜂下颌腺和下咽腺的分泌物。这是一种白色或淡黄色的乳脂状物质，为幼蜂幼虫提供食物(但不超过三天，然后用花粉、花蜜和蜂蜜的混合物喂养) ，是蜂王在幼虫和成虫期的唯一食物[95-97]。这种取食方式的差异被认为是造成工蜂和蜂王发育分化的主要因素。与工人的食物相比，蜂王浆含有更少的水和四倍多的糖，更多的蛋白质和不同浓度的矿物盐。这种蜂王浆的独特成分导致了基因表达的改变(最有可能是通过表观遗传机制) ，例如，允许卵巢完全发育。多亏了蜂王浆，蜂王可以活到5年(工蜂通常活45天) ，每天产2000-3000个卵。为了商业用途，蜂王浆是从蜂王的细胞中收集的，因为它们是这种产品最丰富的来源ーー蜂王浆的产量远远超过蜂王幼虫能够消耗的量。据有关人士透露，王浆年产量达数千吨，仅中国一国就有约2000吨。
4.1. Royal Jelly Composition
From the chemical point of view, the royal jelly is an emulsion of proteins, sugars, and lipids in water. Moreover, it contains about 1.5% mineral salts (mainly copper, zinc, iron, calcium, manganese, potassium, and sodium salts) and small amounts of flavonoids, polyphenols, and vitamins (biotin, folic acid, inositol, niacin, pantothenic acid, riboflavin, thiamine, and vitamin E) [95, 99–101]. Among RJ, flavonoids can be distinguished: flavanones (hesperetin, isosakuranetin, and naringenin), flavones (acacetin, apigenin, and its glucoside, chrysin, and luteolin glucoside), flavonols (isorhamnetin and kaempferol glucosides), and isoflavonoids (coumestrol, formononetin, and genistein) .
从化学角度来看，蜂王浆是水中蛋白质、糖和脂类的乳化物。此外，它还含有大约1.5% 的矿物盐(主要是铜、锌、铁、钙、锰、钾和钠盐)和少量的黄酮、多酚和维生素(生物素、叶酸、肌醇、烟酸、泛酸、核黄素、硫胺和维生素 e)。在 RJ 中，黄酮类化合物可以区分为: 黄酮类化合物(橙皮素、异樱草素和柚皮素)、黄酮类化合物(acacetin、芹菜素及其葡萄糖苷、白杨素和木犀草素葡萄糖苷)、黄酮醇类化合物(异鼠李素和山奈酚葡萄糖苷)和异黄酮类化合物(香豆素、甲醛酮和染料木素)。
The water content in the royal jelly is 50–70% [23, 95]. The total sugar content fluctuates between 7 and 21.2% and mainly consists of fructose and glucose [95, 99, 103]. Fructose and glucose together account for 90% of all sugars [99, 104]. Sucrose is always present but often in variable concentrations (2.86% according to Kanelis et al. , 2.1% according to Kolayli et al. , 0.5–2% according to Oršolić , and 0.2% according to Wytrychowski ). Studies have also revealed the presence of other oligosaccharides, such as trehalose, maltose, gentiobiose, isomaltose, raffinose, erlose, and melezitose [103, 104].
蜂王浆中的水分含量为50-70% [23,95]。总糖含量在7% ー21.2% 之间波动，主要由果糖和葡萄糖组成[95,99,103]。果糖和葡萄糖加起来占所有糖的90% [99,104]。蔗糖总是存在，但经常在不同的浓度(2.86% 根据 Kanelis 等人 ，2.1% 根据 Kolayli 等人 ，0.5-2% 根据 or oli  ，和0.2% 根据 Wytrychowski )。研究还发现了其他低聚糖的存在，例如海藻糖、麦芽糖、龙胆糖、异麦芽糖、棉子糖、麦角糖和多糖[103,104]。
The total protein content in the royal jelly, according to different researchers, varies between 8 and 9% [23, 104, 105]. The electrophoretic analysis of royal jelly from two bee subspecies—Apis cerana japonica and Apis mellifera—revealed 21 different bands of proteins on the gel of which 14 bands were common to both subspecies . The so-called major royal jelly proteins (MRJPs) represent about 90% of the total protein content . Interestingly, according to Kamakura , the ability of the royal jelly to modulate the development of female larvae may be partially related to the presence the most abundant protein—major royal jelly protein 1 (MRJP1). Silici et al.  pointed out that very important components of the royal jelly are free amino acids. Using the LC/MS method, they indicated that it contained amino acids such as lysine (the biggest amount—62.43 mg/100 g); proline (58.76 mg/100 g); cystine (21.76 mg/100 g); aspartic acid (17.33 mg/100 g); and less than 5 mg/100 g of valine, glutamic acid, serine, glycine, cysteine, threonine, alanine, tyrosine, phenylalanine, hydroxyproline, leucine-isoleucine, and glutamine. According to these scientists, the antioxidant activity of royal jelly may be related to the biological effect of free amino acids.
根据不同研究人员的研究，蜂王浆中的总蛋白质含量在8% 到9% 之间。对中华蜜蜂(Apis cerana japonica)和意大利蜜蜂(Apis mellifera)两个蜜蜂亚种的蜂王浆进行电泳分析，发现凝胶上有21条不同的蛋白带，其中14条带与两个亚种相同。所谓的主要蜂王浆蛋白(MRJPs)约占总蛋白质含量的90% 。有趣的是，根据镰仓 ，蜂王浆调节雌性幼虫发育的能力可能部分与存在最丰富的蛋白质ーー主要蜂王浆蛋白1(MRJP1)有关。Silici 等人指出，蜂王浆中非常重要的成分是游离氨基酸。用液相色谱-质谱联用技术(LC/MS)测定，其中赖氨酸(最大含量为62.43 mg/100g)、脯氨酸(58.76 mg/100g)、胱氨酸(21.76 mg/100g)、天冬氨酸(17.33 mg/100g) ，以及少于5mg/100g 的缬氨酸、谷氨酸、丝氨酸、甘氨酸、半胱氨酸、苏氨酸、丙氨酸、酪氨酸、丙氨酸、羟脯氨酸、亮氨酸-异亮氨酸和苯胺。据这些科学家说，蜂王浆的抗氧化活性可能与游离氨基酸的生物效应有关。
The total content of fats and fatty acids in the royal jelly is estimated to be in the range of 7–18% [95, 104]. Instead of carboxylic acids with 14–20 carbon atoms commonly found in animals and plants, the royal jelly contains short hydroxy fatty acids with 8–12 carbon atoms in the chain and dicarboxylic acids. About 80–90% of the fatty substance fraction is an extremely rare free fatty acid with an unusual structure. The major fatty acid is 10-hydroxydecanoic acid (10-HDA), whose presence has not been reported in any other natural raw material or even in any other product of apiculture . According to different investigators, the content of 10-HDA in the royal jelly ranges from 0.75 to 3.39% . It is noteworthy that this acid is considered as one of the most important components from which the royal jelly biological activity derives. Other carboxylic acids are 10-hydroxy-2-decenoic acid (10H2DA) and sebacic acid (SA) (Figure 4) .
据估计，蜂王浆中的脂肪和脂肪酸总含量在7-18% [95,104]。蜂王浆不同于动物和植物中常见的含14-20个碳原子的羧酸，而是含有链中含8-12个碳原子的短羟基脂肪酸和二羧酸。大约80-90% 的脂肪物质部分是一个非常罕见的游离脂肪酸具有不寻常的结构。主要的脂肪酸是10- 羟基癸酸(10-HDA) ，其存在尚未在任何其他天然原料或甚至在任何其他养蜂产品中被报道。根据不同的研究结果，蜂王浆中10-HDA 的含量在0.75ー3.39% 之间。值得注意的是，这种酸被认为是一个最重要的组成部分，从蜂王浆的生物活性来源。其他羧酸是10- 羟基 -2- 癸烯酸(10H2DA)和癸二酸(SA)(图4)。
Considering the antioxidant activity, very important ingredients of the royal jelly are flavonoids and phenolic compounds. According to Nabas et al. , the royal jelly contains 23.3 ± 0.92 GAE μg/mg total of phenolics and 1.28 ± 0.09 RE μg/mg of total flavonoids. Interestingly, Liu et al.  found higher contents of polyphenolic compounds (and also proteins) in the royal jelly harvested 24 hours than in that harvested 48 or 72 hours after collection from the larvae. Hence, the authors suggest that the harvesting time of royal jelly can affect the content of antioxidant compounds and, thus, the therapeutic potential of the product. The GC/MS analysis performed by Kanbur et al.  showed that the main phenolic compounds contained in the royal jelly were pinobanksin as well as organic acids and their esters, for example, octanoic acids, 2-hexenedioic acid and its esters, dodecanoic acid and its ester, 1,2-benzenedicarboxylic acid, and benzoic acid.
考虑到蜂王浆的抗氧化活性，其重要成分是黄酮类和酚类化合物。据 Nabas 等 ，蜂王浆中酚类化合物总量为23.3 ± 0.92 μg/mg，黄酮类化合物总量为1.28 ± 0.09 μg/mg。有趣的是，Liu 等人发现，在蜂王浆收获24小时后，其多酚化合物(以及蛋白质)的含量高于收获48或72小时后的蜂王浆。因此，作者认为，蜂王浆的收获时间可以影响抗氧化剂化合物的含量，从而影响产品的治疗潜力。对蜂王浆中的酚类化合物进行了 GC/MS 分析，结果表明，蜂王浆中的酚类化合物主要是粉碎素和有机酸及其酯类，如辛酸、2- 己二酸及其酯类、十二烷酸及其酯类、1,2- 苯二甲酸和苯甲酸。
4.2. Royal Jelly as a Scavenger of Free Radicals
In the available data, there are some reports confirming the role of royal jelly as a scavenger of free radicals [104, 110, 112, 113]. For instance, Liu et al.  investigated the antioxidant properties of the royal jelly expressed as the radical-scavenging effect upon DPPH, hydroxyl, and superoxide radicals. The researchers also evaluated its reducing power, inhibition effect upon linoleic acid oxidation, and superoxide dismutase activity. The obtained results were compared depending on the larval age (1-, 2-, or 3-day old) and time of harvest after the larval transfer from the queen cell cups to the bee hives (24, 48, and 72 h). The authors noted DPPH radical-scavenging effect (in the range of 43.0–62.8%) as well as the inhibitory effect on the superoxide radical formation (ranging from 23.9 to 37.4%) and on hydroxyl radical formation (48–68%). Moreover, the royal jelly sample demonstrated an inhibitory effect on linoleic acid peroxidation (8.6–27.9%). In all cases, the strongest scavenging effect of RJ was noted in the samples taken from the youngest larvae (1 day old) transferred into bee hives for the shortest time (24 h). In addition, the same royal jelly samples proved to have the strongest reducing power. On the other hand, the SOD activity of the royal jelly collected at 72 h after larval transferring of 3-day old larvae was significantly higher than that of the others. Accordingly, the authors suggested that the superoxide radical-scavenging effect of the royal jelly might be attributed to antioxidative compounds different from SOD.
在现有的数据中，有一些报告证实蜂王浆具有清除自由基的作用[104,110,112,113]。例如，Liu 等人研究了蜂王浆的抗氧化特性，表现为对 DPPH、羟基和超氧自由基的清除作用。研究人员还评估了它的还原能力、对亚油酸氧化的抑制作用和超氧化物歧化酶活性。取得的结果进行了比较，取决于幼虫年龄(1-，2-，或3天)和收获时间的幼虫从蜂王细胞杯转移到蜂巢(24,48，和72小时)。作者注意到 DPPH 自由基清除作用(43.0ー62.8%) ，对超氧阴离子自由基形成的抑制作用(23.9ー37.4%)和对羟自由基形成的抑制作用(48ー68%)。此外，蜂王浆样品对亚油酸过氧化具有抑制作用(8.6-27.9%)。在所有情况下，从最小幼虫(1天大)转移到蜂巢的最短时间(24小时)取样中发现 RJ 的清除作用最强。此外，同样的蜂王浆样品被证明具有最强的还原力。另一方面，3日龄幼虫转移后72小时采集的蜂王浆 SOD 活性明显高于其他幼虫。因此，作者认为蜂王浆清除超氧自由基的作用可能来源于不同于 SOD 的抗氧化化合物。
Guo et al.  found strong antioxidant properties of peptides obtained after the hydrolysis of royal jelly proteins using protease N. The antioxidative properties of the obtained peptides were examined in terms of mechanisms such as hydrogen peroxide, superoxide, and hydroxyl radical-scavenging activities and metal-chelating activity. Twelve obtained peptides showed strong hydroxyl radical-scavenging activity, and three dipeptides containing Tyr residues at their C-termini (Lys-Tyr, Arg-Tyr, and Tyr-Tyr) had strong hydrogen peroxide-scavenging activity. However, in this study, no significant metal-chelating and superoxide anion-radical-scavenging activities of the isolated peptides were noted. The authors concluded that di- and tri-peptides could possess greater antioxidative activity than their constituent amino acids.
郭等人发现，用蛋白酶核磁共振水解蜂王浆蛋白后得到的肽具有强大的抗氧化性能。从过氧化氢、超氧阴离子自由基、羟自由基清除和金属螯合等机理考察了所得多肽的抗氧化性能。所获得的十二种肽具有强的羟基自由基清除活性，其中3种含 Tyr 残基的二肽(lys-Tyr、 arg-Tyr 和 Tyr-Tyr)具有强的过氧化氢清除活性。然而，在这项研究中，没有发现明显的金属螯合和超氧阴离子自由基清除活性的分离肽。作者认为二肽和三肽具有比其组成氨基酸更强的抗氧化活性。
4.3. Antioxidant Effect of Royal Jelly in Human and Animal Diabetes Mellitus Model
Despite antioxidant properties of the royal jelly found in both in vitro and in vivomodels, there are only a few human studies confirming its effectiveness. The research recently conducted concerned its influence on the parameters associated with diabetes and oxidative stress in people with diabetes mellitus type 2 [114, 115]. In the study conducted by Pourmoradian et al. , 50 female volunteers with type 2 diabetes were randomly supplemented with RJ (1000 mg once a day) or placebo for 8 weeks. Before and after the intervention, glycemic and antioxidative-oxidative blood parameters were determined. After the supplementation decreased fasting blood glucose (FBG) and serum glycosylated hemoglobin (HbA1c) levels as well as increased insulin concentration were noticed in the royal jelly-supplemented group in comparison with the placebo one. Moreover, the supplementation caused a significant increase in erythrocyte SOD and GPx activities as well as a decrease in MDA concentration. Similar results were reported by Shidfar et al. . In their study, 46 type 2 diabetic patients were randomly assigned to royal jelly (1000 mg, 3 times a day, for 8 weeks) or placebo -supplemented groups. In the supplemented group, decreased homeostasis model assessment for insulin resistance (HOMA-IR) and increased total antioxidant capacity in comparison with the placebo group were noted. Also in studies using an animal model of diabetes, the improvement of oxidative-antioxidant (MDA, CAT, and ferric-reducing properties of plasma (FRAP)) and biochemical parameters (alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and fasting blood glucose (FBG)) as well as histopathological changes (tubular differentiation index, mononuclear immune cells, tunica albuginea thickness, seminiferous tubules diameter, Johnsen’s score, spermiogenesis index, Sertoli cell index, and meiotic index) were observed after royal jelly supplementation [116, 117]. The authors suggested that their results confirmed the role of reactive oxygen species, even if only secondary, in the pathogenesis of type 2 diabetes. According to them, the royal jelly can ameliorate insulin resistance via antioxidant effect. Based on their results, the authors stated that supplementation with the royal jelly might be beneficial for diabetic patients, but further studies are necessary to clarify the exact mechanism of RJ influence on diabetic parameters.
尽管在体外和体内模型中都发现了蜂王浆的抗氧化特性，但只有少数人类研究证实了它的有效性。这项研究最近关注的是它对糖尿病患者和氧化应激相关参数的影响[114,115]。在 Pourmoradian 等人进行的研究中，50名患有2型糖尿病的女性志愿者随机补充 RJ (每天1000毫克)或安慰剂8周。干预前后测定血糖和抗氧化血液参数。补充蜂王浆组空腹血糖(FBG)和糖化血红蛋白(HbA1c)水平下降，胰岛素浓度增加，与安慰剂组相比。此外，补充剂能显著提高红细胞 SOD 和 GPx 活性，降低 MDA 含量。Shidfar 等人也报道了类似的结果。在他们的研究中，46名2型糖尿病患者被随机分配到蜂王浆(1000毫克，每天3次，持续8周)或安慰剂补充组。在补充组中，与安慰剂组相比，胰岛素抵抗的稳态模型评估(HOMA-IR)下降，总抗氧化能力增加。在糖尿病动物模型的研究中，观察了补充蜂王浆后氧化抗氧化剂(MDA、 CAT 和血浆铁还原特性(FRAP)、生化指标(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)和空腹血糖(FBG)的改善以及组织病理学变化(肾小管分化指数、免疫细胞、卵巢白膜厚度、曲细精管直径、 sen 评分、精子发生指数、 Sertoli 细胞指数和减数分裂指数)。作者认为他们的结果证实了活性氧类在2型糖尿病发病机制中的作用，即使只是继发性的。据他们说，蜂王浆可以通过抗氧化作用改善胰岛素抵抗。基于他们的结果，作者指出补充蜂王浆可能对糖尿病患者有益，但进一步的研究有必要澄清 RJ 对糖尿病参数影响的确切机制。
4.4. Antioxidant and Neuroprotective Effects of Royal Jelly
There are several studies that focused on the relationship between the antioxidant and neuroprotective effects of royal jelly, in the literature data. Mohamed et al.  investigated the possible neurotoxic effect of tartrazine, a commonly used synthetic azo dye, as well as the potential modulatory role of royal jelly. The group of rats receiving only tartrazine showed not only disturbances of antioxidant biomarkers but also numerous apoptotic cells in the brain cortex and significant decrease in the concentration of the brain neurotransmitters (GABA, dopamine, and serotonin). The authors revealed that the cotreatment of rats with royal jelly improved antioxidant biomarkers as well as neurotransmitter levels. Interestingly, royal jelly also had an activating effect on the central nervous system represented by the reduced degree of damage and apoptosis of brain tissue. The authors concluded that a component responsible for these changes could be 10-hydroxy-2-decenoic acid, because it was demonstrated that in addition to its antioxidative properties, 10H2DA could support the generation of neurons.
文献数据表明，蜂王浆的抗氧化作用与神经保护作用之间存在着一定的关系。Mohamed et al. 调查了酒石黄(一种常用的合成偶氮染料)可能的神经毒性作用，以及蜂王浆的潜在调节作用。仅服用柠檬黄的那组大鼠不仅表现出抗氧化剂生物标志物的紊乱，而且大脑皮层中有大量的凋亡细胞，大脑神经递质(GABA、多巴胺和5- 羟色胺)浓度显著降低。研究人员发现，与蜂王浆配合使用可以提高抗氧化剂生物标志物和神经递质水平。有趣的是，蜂王浆对中枢神经系统也有激活作用，表现为减少脑组织的损伤和凋亡。作者得出结论，引起这些变化的成分可能是10- 羟基 -2- 癸烯酸，因为研究表明，除了它的抗氧化特性之外，10H2DA 还可以支持神经元的生成。
The relationship between the neutralizing effect of royal jelly on oxidative stress and neurotoxicity was also sought by Aslan et al. . The researchers revealed that royal jelly diminished the secondary neuronal damage after experimental spinal cord injury in rabbits. In this study, the authors noticed that the treatment with royal jelly prevented lipid peroxidation and augmented endogenous enzymic or nonenzymic antioxidative defense systems levels (Table 4). Moreover, royal jelly treatment significantly decreased the apoptotic cell number induced by spinal cord injury. Because the authors noted statistically higher levels of ascorbic acid in the royal jelly group (laminectomy +100 mg/kg RJ p.o.) in comparison with the control group (laminectomy + single dose of 1 mL/kg saline p.o.), they suggested that the protective effect of royal jelly against oxidative stress might be related to restoration of ascorbic acid availability.
蜂王浆对氧化应激的中和作用和神经毒性之间的关系也被 Aslan 等人研究过。研究人员发现，蜂王浆减少了兔子在实验性嵴髓损伤后继发性神经元损伤。在这项研究中，作者注意到使用蜂王浆的治疗可以防止脂质过氧化和增加内源性酶或非酶的抗氧化系统水平。此外，蜂王浆治疗显著降低了嵴髓损伤诱导的凋亡细胞数量。因为作者注意到蜂王浆组(椎板切除 + 100mg/kg RJ p.o.)与对照组(椎板切除 + 1 mL/kg 生理盐水 p.o.)相比抗坏血酸水平有统计学上的提高，他们提示蜂王浆对氧化应激的保护作用可能与恢复抗坏血酸的可用性有关。
|Source资料来源Toxic/harmful factor 有毒/有害因素Harmful effects of an applied factor 应用因素的有害影响The dose and the way of application of royal jelly or its ingredients 蜂王浆及其成分的用量和使用方法Effects of royal jelly or its ingredient coadministration 蜂王浆及其配方共同使用的效果Neuroprotective effect of royal jelly 蜂王浆的神经保护作用Mohamed et al. [ Mohamed 等人Tartrazine-induced neurotoxicity in rats (500 mg/kg酒石黄致大鼠神经毒性(500mg/kg)p.o.假释官, 30 days) ，30天)↑ MDA; ↓ SOD, CAT, and GSH in brain tissue 4、脑组织中的 SOD、 CAT 和 GSHRJ: 300 mg/kgRJ: 300毫克/千克p.o. 假释官, 30 days 30天↓ MDA; ↑ SOD, CAT, and GSH in brain tissue ↓ MDA; ↑脑组织中的 SOD、 CAT 和 GSHAslan et al. [等人Neuronal damage after experimental spinal cord injury (laminectomy) in rabbits 兔实验性椎板切除术后神经元损伤的嵴髓损伤↓ Nitrate and nitrite in serum 4. 血清中的硝酸盐和亚硝酸盐|
↓ SOD and GPx; ↑ CAT in erythrocytes 红细胞中的 SOD 和 GPx; ↑ CAT
↑ MDA, nitrite, and nitrate, 丙二醛、亚硝酸盐、硝酸盐,
↓ GSH in cerebrospinal fluid 公司的↓ GSH 脑嵴液
↑ MDA and GSH in brain tissue 脑组织中 MDA ↑和 GSH 的表达RJ: 100 mg/kg b.w. RJ: 100毫克/千克 b.wp.o.假释官 after trauma 外伤后↓ MDA, ↑GSH in whole blood ↓ MDA，↑全血中的 GSH
↑ Nitrate, Vit. C, retinol, and维特河，视黄醇，和β-carotene in serum – 血清中胡萝卜素
↑ SOD, CAT, and GPx in erythrocytes 红细胞中 SOD、 CAT 和 GPx
↓ MDA and nitrite; ↑ GSH in cerebrospinal fluid ↓ MDA 和亚硝酸盐; ↑脑嵴液谷胱甘肽
↓ MDA; and ↑ GSH in brain tissue 脑组织↓ MDA 和↑ GSHTeixeira et al. [ 特谢拉等人Resistant and cold stress condition 抗冷应力条件↑ TBARS brain, cerebellum, cerebral cortex, and hippocampus 脑、小脑、大脑皮层和海马
↓ GPx, GR, G6PDH, and GSH in the brain and striatum 在大脑和纹状体中↓ GPx，GR，G6PDH 和 GSHRJ: 200 mg/kg by gavage, 14 days RJ: 灌胃200毫克/千克，14天↓ TBARS level in the brain, cerebellum, striatum, and hippocampus 4、大脑、小脑、纹状体和海马体的 TBARS 水平
↑ GPx, GR, G6PDH, and GSH concentration in cerebral cortex and striatum 大脑皮层和纹状体中 GPx、 GR、 G6PDH 和 GSH 的浓度Inoue et al. [ 井上等人6-Hydroxydopamine- (6OHDA-) induced cell death; human neuroblastoma SH-SY5Y cells 6-Hydroxydopamine-(6OHDA -)诱导细胞死亡人神经母细胞瘤 SH-SY5Y 细胞↑ ROS generation 活性氧调节子的产生RJ fatty acid derivative—HPO-DAEE: 50 RJ 脂肪酸衍生物ー hpo-daee: 50μM↑ Expression of HO-1 mRNA HO-1 mRNA 的↑表达
↑ Cell viability ↑细胞活力
↓ ROS generation 公司名片↓Mitigation effect of royal jelly on chemotherapeutic agents 蜂王浆对化疗药物的缓解作用Silici et al. [ 斯里里里等人Cisplatin-induced spermiotoxicity in rats (7 mg/kg b.w. 顺铂诱导的大鼠精子毒性(7mg/kg b.wi.p.in single dose) 单次剂量)↑ MDA 温度
↓ SOD, CAT, and GPx in testis tissues 4. 睾丸组织中的 SOD、 CAT 和 GPxRJ pretreatment and posttreatment: 50 or 100 mg/kg b.w. RJ 预处理和后处理: 50或100mg/kg b.wp.o. 假释官once a day, for 10 days 一天一次，持续10天↓ MDA 4. ↓ MDA
↑ SOD, CAT, and GPx in testis tissues 睾丸组织中 SOD、 CAT 和 GPxSilici et al. [ 斯里里里等人Cisplatin-induced nephrotoxicity in rats (7 mg/kg 顺铂所致大鼠肾毒性(7mg/kgi.p. in single dose) 单次剂量)↑ MDA 温度
↓ SOD, CAT, and GPx in renal tissues 4. 肾组织中的 SOD，CAT 和 GPxRJ pretreatment and posttreatment: 50 or 100 mg/kg b.w. RJ 预处理和后处理: 50或100mg/kg b.wp.o. 假释官once a day, for 10 days 一天一次，持续10天↓ MDA 4. ↓ MDA
↑ SOD, CAT, and GPx in renal tissues 肾组织中 SOD、 CAT 和 GPx 的表达Amirshahi et al. [ 阿米尔沙希等Bleomycin-induced spermiotoxicity in rats (10 mg/kg b.w., 48 days, twice a week, 博莱霉素致大鼠精子毒性(10mg/kg b.w. ，48 d，每周2次，每次1次)i.p.)↑ MDA in testicular tissue 睾丸组织中 MDA ↑RJ: 100 mg/kg b.w., RJ: 100毫克/千克 b.w. ,p.o. 假释官, 48 days 48天↓ MDA in testicular tissue 睾丸组织中的 MDAKaynar et al. [等人Methotrexate-induced oxidative stress in rats (20 mg/kg b.w. 甲氨蝶呤诱导大鼠氧化应激(20mg/kg b.wi.p., single dose) 、单次剂量)↑ MDA and ↓ SOD and GPx in plasma 血浆中的酶活性和↓ SOD 和 GPxRJ: 50 or 100 mg/kg b.w., RJ: 50或100毫克/千克 b.w. ,p.o, 10 days 10天↓ MDA and ↑ SOD ↓ MDA 和↑ SOD∗∗ 第 x 次/次 and GPx 还有 GPx∗∗ 第 x 次/次 in plasma 等离子体Malekinejad et al. [ 马来贾德等人127]Paclitrexal-induced cardiotoxicity in rats (7.5 mg/kg b.w. Paclitrexal 致大鼠心脏毒性(7.5 mg/kg b.wi.p., weekly, 7 weeks) ，每周，7星期)↓ TAC in serum 4. 血清中的 TAC
↑ MDA and NO in heart tissue 心肌组织中 MDA ↑和 NORJ: 50, 100, or 150 mg/kg b.w.,RJ: 50,100，或150毫克/千克 b.w. ,p.o. 假释官, 28 days 28天↑ TAC∗∗ 第 x 次/次 in serum在血清中
↓ MDA 4. ↓ MDA∗∗ 第 x 次/次 and NO in heart tissue 和心脏组织中的一氧化氮Delkhoshe-Kasmaie et al. [ Delkhoshe-Kasmaie 等Taxol-induced damage of the testis (7.5 mg/kg b.w. 紫杉醇诱导的睾丸损伤(7.5 mg/kg b.wi.p., weekly, 4 weeks) 、每周、4星期)↑ MDA and NO and ↓ TTM in testis tissue 精巢组织中的 MDA ↑、 NO 和 TTM ↓RJ: 50, 100, or 150 mg/kg b.w., 4 weeks RJ: 50,100，或150毫克/千克 b.w. ，4周↓ MDA and NO 4. ↓ MDA 和 NO∗∗ 第 x 次/次 and ↑ TTM 和↑ TTM∗∗ 第 x 次/次 in testis tissue 在睾丸组织中Mitigation effect of royal jelly on other toxic agents 蜂王浆对其他有毒物质的缓解作用Kanbur et al. [等人.Sodium fluoride-induced oxidative stress in mice (200 ppm fluoride 氟化钠诱导小鼠的氧化应激(200ppm 氟化物p.o. 假释官, 7 days) ，7天)↑ MDA in erythrocytes and liver tissue 红细胞和肝组织中 MDA ↑
↓ SOD, CAT, and GPx in erythrocytes 4. 红细胞中的 SOD、 CAT 和 GPx
↑ GPx, ↓ CAT, and SOD in the liver tissue 4. 肝组织中的氧化酶RJ: 50 mg/kg b.w. by gavage for 7 days RJ: 50毫克/千克每小时，灌胃7天↓ MDA in erythrocytes and liver tissue 4. 红细胞和肝组织中的 MDA
↑ SOD and CAT in erythrocytes and liver tissue 红细胞和肝组织中的↑ SOD 和 CAT
↓ GPx in erythrocytes 4. 红细胞↓ GPxCemek et al. [等人Carbon tetrachloride-induced acute liver damage in rats (0.8 mL/kg b.w. 四氯化碳致大鼠急性肝损伤(0.8 mL/kg b.ws.c. 南卡罗来纳, 20 days) 20天)↑ MDA in the whole blood, liver, brain, kidney, lung, and heart tissues 全血、肝、脑、肾、肺、心组织中 MDA ↑
↓ GSH in the whole blood ↓全血中的谷胱甘肽
↓ Vit. C, 4. ↓,β-carotene, and retinol in serum – 胡萝卜素及视黄醇RJ: 50, 100, and 200 mg/kg b.w., RJ: 50,100，和200毫克/千克 b.w. ,p.o. 假释官, 20 days 20天↓ MDA in the whole blood, liver 4. 全血、肝脏中的 MDA∗∗ 第 x 次/次, brain ，大脑∗∗ 第 x 次/次, kidney, lung, and heart tissues 肾，肺，心脏组织
↓ GSH in the whole blood ↓全血中的谷胱甘肽∗∗ 第 x 次/次
↑ GSH in the liver and brain tissues 脑组织和肝组织中的↑型谷胱甘肽∗∗ 第 x 次/次
↑ Vit. C, ↑,β-carotene, and retinol in serum – 胡萝卜素及视黄醇Ahmed et al. [ 艾哈迈德等124]Azathioprine-induced toxicity in rats (50 mg/kg b.w. 硫唑嘌呤(azathioprine)对大鼠(50mg/kg b.wi.p., single dose) 、单次剂量)↑ MDA and GSH in the liver tissue 肝组织中 MDA ↑和 GSH 的表达RJ: 200 mg/kgRJ: 200毫克/千克p.o. 假释官, 7 days 7天↓ MDA and ↑ GSH in the liver tissue after 24 h and 2 weeks of posttreatment ↓ MDA 和↑ GSH 在治疗后24小时和2周肝组织中的表达Ghanbari et al. [ 甘巴里等人Streptozotocin-induced diabetes mellitus (60 mg/kg b.w., 链脲佐菌素诱导的糖尿病(60mg/kg b.w. ,i.p.)↑ MDA, ↓ CAT, and FRAP in the liver and pancreas 4、肝脏和胰腺中的↓ CAT 和 FRAPRJ: 200 mg/kg b.w., RJ: 200毫克/千克 b.w. ,p.o. 假释官,
6 weeks 6周↓ MDA in the liver and pancreas 肝脏和胰腺中的 MDA
↑ CAT and FRAP in the liver and pancreas 肝脏和胰腺中 CAT 和 FRAP 的表达Ghanbari et al. [ 甘巴里等人Streptozotocin-induced diabetes mellitus (50 mg/kg b.w., 链脲佐菌素诱导的糖尿病(50mg/kg b.w. ,i.p.)↓ CAT and FRAP in testicular tissue 睾丸组织的↓ CAT 和 FRAPRJ: 200 mg/kg b.w., rJ: 200毫克/千克 b.w. ,p.o. 假释官,
6 weeks 6周↑ CAT and FRAP in the testicular tissue 睾丸组织中↑ CAT 和 FRAP 的表达Sugiyama et al. [ 杉山等人LPS- and interferon- lPS 和干扰素β-induced NO generation; RAW264 murine macrophage cell line 诱导 NO 产生; RAW264鼠巨噬细胞系↑ Nitrate↑硝酸盐
↑ iNOS promoter activity iNOS ↑启动子活性
↑ NF-κB activation and TNF- B 激活与肿瘤坏死因子-αproduction生产RJ fatty acid (1 mM, 2 mM, 4 mM 10H2DA) RJ 脂肪酸(1mm，2mm，4mm，10H2DA)↓ Nitrate ↓硝酸盐
↓ iNOS promoter activity 4. iNOS ↓启动子活性
↓ NF- ↓-κB activation and TNF- B 激活和 TNF-αproduction 生产Takahashi et al. [ 高桥等人Interferon- 干扰素-γ-induced NO production; RAW264 murine macrophage cell 小鼠巨噬细胞系 RAW264↑ Nitrate↑硝酸盐
↑ iNOS promoter activation iNOS 基因座↑启动子激活
↑ NF-κB activation and TNF- B 激活和 TNF-αproduction生产RJ fatty acid (1 mM, 2 mM, 5 mM 10H2DA) RJ 脂肪酸(1mm，2mm，5mm，10H2DA)↓ Nitrate ↓硝酸盐
↓ iNOS promoter activation and NF- iNOS ↓启动子激活和 NF-κB activation B 激活∗∗ 第 x 次/次 and TNF- 肿瘤坏死因子-αproduction 生产
|10H2DA: 10-hydroxy-trans-2-decenoic acid; CAT: catalase; FRAP: iron reduction capacity; G6PDH: glucose-6-phosphate dehydrogenase; GPx: glutathione peroxidase; GR: glutathione reductase; GSH: reduced glutathione; G6PDH: glucose-6-phosphate dehydrogenase; HPO-DAEE: hydroperoxy-2-decenoic acid ethyl ester; HO-1: heme oxygenase-1; iNOS: inducible nitric oxide synthase; MDA: malondialdehyde; NO: nitric oxide; NF- 10H2DA: 10- 羟基反式 -2- 癸烯酸; CAT: 过氧化氢酶; FRAP: 铁还原能力; G6PDH: 葡萄糖-6-磷酸脱氢酶; GPx: 谷胱甘肽过氧化物酶; GR: 谷胱甘肽还原酶; GSH: 谷胱甘肽; G6PDH: 葡萄糖-6-磷酸脱氢酶; HPO-DAEE: 氢过氧化2- 癸烯酸乙酯; HO-1: 血红素氧合酶 -1; iNOS: 诱导醛; MDA: 丙二醛; NO: 氧化物;-κB: nuclear factor kappa-light-chain-enhancer of activated B cells; Nrf2/ARE: nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant responsive elements (AREs); SOD: superoxide dismutase; TAC: total antioxidant capacity; TBARS: thiobarbituric acid reactive substances; TNF- B: 活化 b 细胞核因子 kappa-light-chain-enhancer; Nrf2/ARE: nuclear factor erythroid related factor 2(Nrf2)/antioxidresponsive elements (AREs) ; SOD: 超氧化物歧化酶; TAC: total antioxidance capacity; TBARS: thiobbitumoic acid reactive substances; TNF-α: tumor necrosis factor alpha; TTM: total thiol molecules; ↓: decrease; ↑: increase; 总巯基分子↓ : 减少; ↑ : 增加; ↑ : 肿瘤坏死因子-α∗∗ 第 x 次/次the effect depended on used dose. 效果取决于使用剂量|
Table 4 表四The protective effects of royal jelly against prooxidant action of different harmful factors. 蜂王浆对不同有害因子促氧化作用的保护作用
A study conducted by Teixeira et al.  also suggested the existence of antioxidant and neuroprotective effects of royal jelly but in the resistant and cold stress condition (Table 4). The authors postulated that the antioxidant activity of royal jelly, observed in the region of striatum, might correspond to adenosine monophosphate (AMP) N1-oxide—the unique compound of RJ—which can regulate neuronal functions through receptors predominantly expressed in striatum (A2A adenosine receptors). They speculated that the activation of these receptors can prevent radical formation and apoptosis.
Teixeira 等人进行的一项研究也表明蜂王浆具有抗氧化和神经保护作用，但是在抗寒和冷应激条件下(表4)。作者推测，在纹状体区域观察到的蜂王浆的抗氧化活性可能与一氧化二单磷酸腺苷(AMP) n1-oxide 相对应，这是 rj 的独特化合物，通过纹状体中主要表达的受体(A2A 腺苷受体)调节神经元功能。他们推测这些受体的激活可以阻止自由基的形成和细胞凋亡。
4.5. Alleviating Effect of Royal Jelly on Oxidative Stress
Studies evaluating the alleviating effect of royal jelly on oxidative stress were performed using several in vivo models. The antioxidant effect of royal jelly in cisplatin-induced spermiotoxicity and nephrotoxicity in rats was investigated by Silici et al. [100, 121]. The researchers attributed antioxidant properties of royal jelly to the presence of substances such as 10-hydroxy-2-decenoic acid and free amino acids including proline (which is suggested to act as an antioxidant due to hydroxyl radical-scavenging activity) as well as cystine and cysteine (participating in the synthesis of effective cellular antioxidant—glutathione). The influence of royal jelly on adverse effects generated by the administration of sodium fluoride at high doses in mice was assessed by Kanbur et al. . The authors explained that the antioxidant royal jelly effect could be associated not only with radical-scavenging effect but also with another indirect effect based on the inhibition of enzymes that catalyze the peroxidation of endogenous lipids as well as the gene expression of cytochrome P450, which is one of the intracellular source of H2O2, O2·− and HO· radicals . Other studies also demonstrate the antioxidant effect of royal jelly in an animal model under oxidative stress condition induced by substances like carbon tetrachloride (industrial solvent) , azathioprine (immunosuppressive drug) , bleomycin , methotrexate , paclitaxel , taxol  (chemotherapeutic agents), and oxymetholone (synthetic androgen analogue) . In all mentioned studies, the antioxidant effect of royal jelly consisted of a positive effect on the oxidative-antioxidative parameters (Table 4). In addition, some researchers reported other therapeutic effects of royal jelly such as hepatoprotective [123, 124], cardioprotective , or anti-inflammatory [113, 130] ones. The postulated hypotheses explaining the antioxidant effect are the restoration of ascorbic acid availability by royal jelly, regulation of retinol loss , antioxidant effect of some free amino acids , or radical-scavenging activities of RJ and its component .
用几种体内模型评价蜂王浆对氧化应激的缓解作用。用 siliii 等[100,121]研究蜂王浆对顺铂所致大鼠精子毒性和肾毒性的抗氧化作用。研究人员将蜂王浆的抗氧化特性归因于存在的物质，如10- 羟基 -2- 癸烯酸和游离氨基酸，包括脯氨酸(由于羟基自由基清除活性被认为是一种抗氧化剂) ，以及胱氨酸和半胱氨酸(参与合成有效的细胞抗氧化剂ー谷胱甘肽)。蜂王浆对小鼠高剂量氟化钠所产生的不良反应的影响是由 Kanbur 等人评估。作者解释了抗氧化蜂王浆效应不仅与自由基清除作用有关，而且还与抑制内源性脂质过氧化的酶以及细胞色素 P450基因表达有关，后者是 H2O2、 O2和 HO 自由基的胞内来源之一。其他研究也证明了蜂王浆在四氯化碳(工业溶剂)、硫唑嘌呤(免疫抑制药)、博来霉素、甲氨蝶呤、紫杉醇、紫杉醇(化疗药物)和氧甲硅酮(合成雄激素类似物)诱导的动物模型中的抗氧化作用。在所有提到的研究中，蜂王浆的抗氧化作用包括对氧化-抗氧化参数的积极影响(表4)。此外，一些研究人员还报道了蜂王浆的其他治疗作用，例如保护肝脏、保护心脏或抗炎作用。解释抗氧化作用的假设是蜂王浆恢复抗坏血酸的有效性，调节视黄醇丢失 ，一些游离氨基酸的抗氧化作用，或 RJ 及其成分的自由基清除活性。
The antioxidant effect of royal jelly has also been confirmed by in vitro studies. For instance, the aim of the study by Inoue et al.  was to investigate protective effects of royal jelly fatty acid derivative (4-hydroperoxy-2-decenoic acid ethyl ester (HPO-DAEE)) on oxidative stress-induced cell death using human neuroblastoma SH-SY5Y cells (Table 4). The researchers noted that the pretreatment with HPO-DAEE protected against 6-hydroxydopamine- (6OHDA-) induced cell death by increasing the expression of antioxidant enzyme—heme oxygenase-1 (HO-1) mRNA—through Nrf2-ARE signaling. Interestingly, the authors revealed that the treatment with HPO-DAEE rapidly induced reactive oxygen species generation in SHSY5Y cells. In conclusion, these results suggested that sublethal oxidative stress caused by HPO-DAEE is essential for the activation of this pathway, which is aimed at antioxidant defense. Moreover, the authors observed that HPO-DAEE promoted the phosphorylation of eukaryotic initiation factor 2a (eIF2a), and the subsequent nuclear accumulation of the activating transcription factor-4 (ATF4). The ATF4 pathway is known to be activated under several stress conditions. It is supposed that the interaction of the Nrf2-ARE pathway with the eIF2a-ATF4 pathway augments HO-1 expression.
蜂王浆的抗氧化作用也得到了体外研究的证实。例如，Inoue 等人的研究目的是探讨蜂王浆脂肪酸衍生物(4- 氢过氧 -2- 癸烯酸乙酯(HPO-DAEE))对人神经母细胞瘤 SH-SY5Y 细胞氧化应激诱导的细胞死亡的保护作用(表4)。研究人员注意到，HPO-DAEE 预处理通过增加抗氧化酶ー血红素氧合酶 -1(HO-1) mrna 的表达，通过 Nrf2-ARE 信号转导，对6- 羟基多巴胺(6OHDA -)诱导的细胞死亡起到保护作用。有趣的是，作者发现 HPO-DAEE 治疗能迅速诱导 SHSY5Y 细胞产生活性氧类。总之，这些结果表明 HPO-DAEE 引起的亚致死性氧化应激对于这一途径的激活是必不可少的，这一途径的目的是为了抗氧化防御。此外，作者观察到 HPO-DAEE 促进了真核起始因子2 a (eIF2a)的磷酸化，以及随后的转录激活因子 -4(ATF4)的核聚集。ATF4通路已知在几种应力条件下被激活。我们认为 Nrf2-ARE 通路与 eIF2a-ATF4通路的相互作用增加了 HO-1的表达。
The investigation of protective effect of royal jelly in the redox state of ovine oocytes matured in vitro, and embryonic development following in vitrofertilization was performed by Eshtiyaghi et al. . The authors explained that the improvement of oocyte maturation in the case of cells supplemented with royal jelly might be associated with the improvement of redox status. One of the objectives of this study was to examine the effect of different concentrations of royal jelly (2.5, 5, and 10 mg/mL of maturation media) on the in vitro maturation and glutathione (GSH) level of ovine oocyte as well as the abundance mRNA of antioxidant enzymes in both oocyte and cumulus cells. Moreover, the authors investigated glucose metabolism-related genes in cumulus cells. The parameters were evaluated following 24 hours of in vitromaturation. The authors noted that the dose of 10 mg/mL of RJ not only led to an increase in the number of oocytes but also caused an increase in the intracellular GSH content compared to the control group and the group receiving the lowest dose of royal jelly. In addition, supplementation with 10 mg/mL of royal jelly increased the mRNA GPx in both oocyte and cumulus cells as well as SOD expression in the cumulus cells. However, royal jelly supplementation did not influence mRNA CAT level in both oocyte and cumulus cells. Moreover, the increased expression of phosphofructokinase and glucose 6-phosphate dehydrogenase in the cumulus cells after the addition of royal jelly to the maturation media indicated that the observed protective effect of royal jelly might be related to the activation of glucose metabolic pathways in the surrounding cumulus cells.
研究了蜂王浆对体外成熟的绵羊卵母细胞氧化还原状态的保护作用，以及对体外人工受精后胚胎发育的保护作用。作者认为，添加蜂王浆的卵母细胞成熟度的提高可能与氧化还原状态的改善有关。研究了蜂王浆(2.5、5、10mg/ml)对绵羊卵母细胞体外成熟和谷胱甘肽(GSH)水平的影响，以及卵母细胞和卵丘细胞中抗氧化酶 mRNA 丰度的影响。此外，作者还研究了卵丘细胞中的葡萄糖代谢相关基因。在体外成熟24小时后对这些参数进行评估。作者注意到，与对照组和蜂王浆最低剂量组相比，10mg/ml 的 RJ 不仅使卵母细胞数量增加，而且使细胞内 GSH 含量增加。此外，添加10mg/ml 蜂王浆可增加卵母细胞和卵丘细胞中 GPx 的 mRNA 表达和卵丘细胞中 SOD 的表达。补充蜂王浆对卵母细胞和卵丘细胞的过氧化氢酶 mRNA 水平均无影响。此外，在成熟培养基中添加蜂王浆后，卵丘细胞中磷酸果糖激酶和葡萄糖-6-磷酸脱氢酶的表达增加，表明蜂王浆的保护作用可能与周围卵丘细胞中葡萄糖代谢途径的激活有关。
The negation of the above results seems to be a study carried out by Filipič et al. . The purpose of this study was to investigate the influence of royal jelly and its bioactive component—10H2DA—and human interferon-alpha (HuIFN-αN3—a protein with antiviral, antiproliferative, and antitumor activities—on the proliferation of human colorectal adenocarcinoma cells (CaCo-2) and the oxidative stress parameters—GSH and MDA concentration. Royal jelly and HuIFN-αN3 applied at 2 : 1 ratio and royal jelly applied in combination with 10H2DA (2 : 1 ratio) caused a decrease in the level of GSH and increase in lipid peroxidation indicator level (MDA) in CaCo-2 cells in comparison with the control group. On the other hand, it was observed that these combinations had the highest antiproliferative effect. The authors suggested that antiproliferative effects of RJ, HuIFN-αN3, and 10H2DA on the CaCo-2 cells could be connected not only with the induction of apoptosis and cytotoxicity but also with their influence on the prooxidative-antioxidative balance.
对上述结果的否定似乎是由 filipi 等人进行的研究。本研究旨在探讨蜂王浆及其生物活性成分ー10h2da ー和人 α 干扰素(huifn ー αn3ー a 抗病毒、抗增殖、抗肿瘤活性蛋白ー对人大肠腺癌细胞(CaCo-2)增殖及其氧化应激参数ー gsh 和 MDA 浓度的影响。与对照组相比，蜂王浆和 huifn αn3以2:1的比例施用，蜂王浆与10H2DA (2:1的比例)联合施用，可使 CaCo-2细胞 GSH 水平降低，MDA 脂质过氧化水平升高。另一方面，观察到这些联合具有最高的抗增殖作用。作者认为 RJ、 HuIFN-αN3和10H2DA 对 CaCo-2细胞的抗增殖作用不仅与诱导凋亡和细胞毒性有关，而且与其对过氧化 — 抗氧化平衡的影响有关。
Attempts to explain the alleviating mechanism of royal jelly’s action on nitrosative stress were made by Sugiyama et al. . Researchers examined the ability of 10H2DA to inhibit LPS-induced nitric oxide (NO) generation using the RAW264 murine macrophage cell line. Their study was based on the fact that LPS stimulates the production of interferon- (IFN-) β, induction of IFN regulatory factor-1, and activation of IFN-stimulated response element. These factors are required for iNOS (nitric oxide synthases) induction. The authors noted that 10H2DA not only inhibited LPS-induced nitric oxide (NO) generation but also restrained IFN-β-induced nuclear factor- (NF-) κB activation and tumor necrosis factor- (TNF-) α production. The authors concluded that 10H2DA inhibited LPS- and IFN-β-induced NO productions via the inhibition of NF-κB activation induced by LPS or IFN-β. Similar studies have been carried out by Takahashi et al. . Using the same cell line, authors noted the inhibition of interferon-γ-induced NO production by 10H2DA through the inhibition of interferon regulatory factor-8 induction (Table 4).
杉山等试图解释蜂王浆对硝化应激的缓解机制。研究人员利用 RAW264小鼠巨噬细胞系检测了10H2DA 抑制 lps 诱导的一氧化氮(NO)生成的能力。他们的研究是基于 LPS 刺激干扰素 β 的产生，干扰素调节因子1的诱导和干扰素刺激反应元件的激活。这些因子是诱导 iNOS (一氧化氮合酶)所必需的。作者指出，10H2DA 不仅抑制 lps 诱导的一氧化氮(NO)的产生，而且还抑制了 ifn-β 诱导的核因子 κb 活化和肿瘤坏死因子-α-(TNF -) α 的产生。10H2DA 通过抑制 LPS 和 ifn β 诱导的 NF-κB 活化，抑制 LPS 和 ifn β 诱导的 NO 产生。Takahashi 等人也进行了类似的研究。利用同一细胞系，作者注意到10H2DA 通过抑制干扰素调节因子 -8的诱导而抑制 γ- 干扰素诱导的一氧化氮生成(表4)。
5. Comparison of the Antioxidant-Related Potential of Propolis, Bee Pollen, and Royal Jelly
Numerous in vitro studies using DPPH, ABTS+, FRAP, ORAC methods, and so on have confirmed the antioxidant potential of bee products [3, 19, 20, 31, 35, 36, 38, 79, 81, 85, 110]. In this part, we will try to compare the antioxidant capacity of propolis, bee pollen, and royal jelly among each other.
利用 DPPH、 ABTS + 、 FRAP、 ORAC 等方法进行的大量体外研究证实了蜂产品的抗氧化潜力[3,19,20,31,35,36,38,79,81,85,110]。在这一部分，我们将尝试比较蜂胶、蜂花粉和蜂王浆之间的抗氧化能力。
It is well known that the antioxidant capacity of bee products is strongly dependent on their chemical composition. In general, the antioxidant activity of poplar propolis is believed to be largely influenced by both total polyphenol and total flavonoid contents, while the Brazilian one by phenolic compounds but different ones than flavonoids [3, 34–36, 38]. In the case of bee pollen, research has shown divergent results; some have shown a strong positive correlation between the total content of phenolic compounds and antioxidant capacity [79, 81], whereas others have found no considerable relationships [71, 87, 136]. Leja et al.  suggested, in turn, that not phenolic compounds but bee pollen phenylpropanoids are responsible for the inhibition of linoleic acid peroxidation. As for royal jelly, its antioxidant properties are mainly attributed to the presence 10-hydroxydecanoic acid and free amino acids including proline as well as cystine and cysteine following phenolic compounds [100, 102, 121].
众所周知，蜂产品的抗氧化能力很大程度上取决于它们的化学成份。一般认为杨木蜂胶的抗氧化活性主要受总多酚和总黄酮含量的影响，而巴西蜂胶的抗氧化活性主要受酚类化合物的影响，但不同于黄酮类化合物[3,34-36,38]。以蜂花粉为例，研究表明了不同的结果; 一些研究表明，总酚类化合物含量与抗氧化能力之间存在很强的正相关性[79,81] ，而另一些研究则没有发现相当大的关系[71,87,136]。Leja 等人提出，反过来，不是酚类化合物而是蜂花粉苯丙素负责抑制亚油酸过氧化作用。至于蜂王浆，其抗氧化特性主要是由于酚类化合物[100,102,121]之后存在10- 羟基癸酸和游离氨基酸，包括脯氨酸、胱氨酸和半胱氨酸。
Basing on literature data, propolis seems to be the most powerful antioxidant among all the analyzed bee products. The comparison of the phenolic compounds’ content seems to confirm the above thesis—the highest amount of both total phenols and flavonoids has been found in propolis followed, in order, by bee pollen and royal jelly. However, it should be emphasized that royal jelly also contains other compounds possessing antioxidant character. The next confirmation of the above thesis are the results obtained by Nakajima et al. . In their study, the rank order of antioxidant potencies measured by the hydrogen peroxide, superoxide anion, and hydroxyl radical-scavenging capacities was as follows: water propolis extract, ethanol propolis extract, and ethanol pollen extract, but neither royal jelly nor 10-hydroxy-2-decenoic acid (10-HDA) had any effects. So both propolis extracts showed much greater antioxidant activity than the bee pollen one, and surprisingly, the water propolis extract was much more effective that the ethanol one.
根据文献资料，蜂胶似乎是所有分析蜂产品中最有效的抗氧化剂。酚类化合物含量的比较似乎证实了上述论点ー蜂胶中总酚类和黄酮类化合物含量最高，其次是蜂花粉和蜂王浆。但应该强调的是，蜂王浆还含有其他具有抗氧化特性的化合物。上述论点的下一个证实是中岛等人的结果。在他们的研究中，抗氧化剂的抗氧化能力测定的过氧化氢，超氧阴离子和羟基自由基清除能力顺序如下: 水蜂胶提取物，乙醇蜂胶提取物，和乙醇花粉提取物，但没有任何影响蜂王浆和10-羟基 -2- 癸烯酸(10-HDA)。两种蜂胶提取物的抗氧化活性都比蜂花粉提取物强得多，令人惊讶的是，水蜂胶提取物的抗氧化活性比乙醇提取物强得多。
On the other hand, in the in vivo study, waterborne bee products (including royal jelly, bee pollen, and propolis) were shown to be able to reverse the oxidative damage caused by exposure to tebuconazole. They affected the brain, kidney, and liver lipid peroxidation, protein carbonylation, and antioxidant markers. All studied products were effective, but the effects were dependent on doses and organs, and no clear trend was observed . Similar results were observed by Turkish researchers  who noticed the highest antioxidant capacity as well as total phenolic and flavonoid content in propolis, followed, in order, by pollen, honey, and royal jelly. Despite very significant differences in both phenolic content and antioxidant activities measured using FRAP and DPPH methods, in an in vivo study, all products revealed similar hepatoprotective activity against CCl4-induced hepatic damage in rats. All of them had the very similar effect on liver parameters and antioxidant/oxidant markers with only a very slight advantage in the case of propolis. The authors suggested that these results could be explained through their bioavailability to the treated animals. It should be underlined that in the case of bee pollen, research has shown not only its antioxidative but also its prooxidative action. For example, in LPS-stimulated macrophages, the bee pollen extract was shown to efficiently scavenge nitric oxide, although against superoxide, it behaved as antioxidant at lower concentrations and as prooxidant at higher concentrations . To the best of our knowledge, prooxidative properties of propolis and royal jelly have not been found. On the other hand, some studies have found that bee pollen along with propolis exhibits strong antioxidant effects, while royal jelly had no effect.
另一方面，在体内研究中，水性蜂产品(包括蜂王浆、蜂花粉和蜂胶)被证明能够逆转暴露于戊唑醇所造成的氧化损伤。它们影响了大脑、肾脏和肝脏的脂质过氧化、蛋白质羰基化和抗氧化标记物。所有研究的产品都是有效的，但效果取决于剂量和器官，没有明显的趋势观察到。土耳其研究人员也观察到类似的结果，他们注意到蜂胶中抗氧化能力最强，总酚和黄酮含量最高，其次是花粉、蜂蜜和蜂王浆。尽管使用 FRAP 和 DPPH 方法测定的酚类物质含量和抗氧化活性有很大的差异，但在一项体内研究中，所有产品都显示出对四氯化碳引起的大鼠肝损伤具有相似的肝保护活性。它们对肝脏参数和抗氧化/氧化标志物的影响非常相似，对蜂胶的影响则非常微弱。作者认为，这些结果可以通过对治疗动物的生物利用度来解释。需要强调的是，研究表明蜂花粉不仅具有抗氧化作用，还具有促氧化作用。例如，在 lps 刺激的巨噬细胞中，蜂花粉提取物被证明能有效清除一氧化氮，尽管它对抗超氧化物，在低浓度时表现为抗氧化剂，在高浓度时表现为促氧化剂。据我们所知，蜂胶和蜂王浆的蜂胶氧化特性还没有被发现。另一方面，一些研究发现蜂花粉和蜂胶具有很强的抗氧化作用，而蜂王浆没有作用。
As for the mechanism underlying the potential antioxidant-related effects, the most studied is propolis. It has been shown the treatment with propolis and its common compound pinocembrin generates an increase in at least one of the enzymatic antioxidant pathway, namely, it induces the translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) to the nucleus and subsequent expression of antioxidant response element- (ARE-) mediated antioxidant genes such has HO-1 and γ-GCS [6, 62]. Moreover, propolis or pinocembrin was found to regulate the expression, on both the mRNA and protein levels, of genes encoding other antioxidant markers, including LOX-1, γ-GCS, GCLM, GCLC, and TrxR1 [6, 49, 56, 63, 66]. The royal jelly was noticed to affect HO-1 expression through Nrf2-ARE pathway as well as the expression of cytochrome P450, GPx, and SOD [122, 131, 132]. As for bee pollen, to the best of our knowledge, there is no information in the literature data on its influence on the expression of antioxidant-related genes. Furthermore, the mechanisms of bee products for the reversal of oxidative damage appear to involve the reduction of lipid peroxidation (MDA and TBARS) and oxidant parameters (e.g., ROS) as well as the augmentation of antioxidant enzyme activities (e.g., CAT, SOD, GPx, and GST). But this assumption is based on the fact that the treatment with bee products affects the level of the above parameters in blood and studied organs.
至于潜在的抗氧化作用的机制，研究最多的是蜂胶。研究表明，蜂胶及其普通化合物松黄素至少增加了一条酶促抗氧化途径，即诱导核因子红系2相关因子2(Nrf2)向细胞核移位，并随后表达抗氧化反应元件-(ARE -)介导的抗氧化基因，如 HO-1和 γ-gcs [6,62]。此外，还发现蜂胶或 pinocemrin 在 mRNA 和蛋白水平上调节其他抗氧化标记基因的表达，包括 LOX-1、 γ-gcs、 GCLM、 GCLC 和 TrxR1[6,49,56,63,66]。蜂王浆通过 Nrf2-ARE 途径影响 HO-1的表达，并影响细胞色素 P450、 GPx 和 SOD [122,131,132]的表达。至于蜂花粉，据我们所知，没有文献资料表明它对抗氧化相关基因表达的影响。此外，蜜蜂产品逆转氧化损伤的机制似乎包括降低脂质过氧化(MDA 和 TBARS)和氧化剂参数(如 ROS) ，以及增加抗氧化酶活性(如 CAT、 SOD、 GPx 和 GST)。但这一假设是基于蜂产品治疗影响血液和研究器官中上述参数的水平这一事实。
Propolis is also the most studied in cell and animal research. It is credited with having antioxidant–related neuroprotective and cardioprotective actions and is thus suggested as a protective agent against Alzheimer’s [47, 48] and Parkinson’s diseases  as well as atherosclerosis . The neuroprotective effect of royal jelly was also noted. All analyzed bee products have been tested in relation to the reduction of the negative effect of chemotherapy—in each case, the obtained results were promising [4, 5, 51, 90, 94, 121, 125, 127, 128]. The rest studies are focused on estimating the protective action of bee products against various harmful factors and drugs causing oxidative stress.
Although in vitro and animal studies seem to confirm the antioxidant-related protective effect of bee products, there are only a few studies performed on humans in the literature data [44–46, 114, 115]. The existing ones aim at evaluating the supplementation effect in healthy population or type 2 diabetic patients and involve propolis as well royal jelly. Herein, the obtained results are inconsistent. For example, propolis supplementation had a positive effect in men but not in women . As for diabetic patients, royal jelly seems to have a higher potential than propolis as it was shown to regulate the parameter associated with diabetes HOMA-IR , while propolis affected only oxidative-related parameters .
尽管体外和动物研究似乎证实了蜂产品的抗氧化相关保护作用，但文献数据中只有少数针对人类的研究[44-46,114,115]。现有的研究主要是评价蜂胶和蜂王浆对健康人群和2型糖尿病患者的补充效果。在此，得到的结果是不一致的。例如，蜂胶补充剂对男性有积极作用，但对女性没有。对于糖尿病患者来说，蜂王浆似乎比蜂胶有更高的潜力，因为蜂王浆可以调节与糖尿病相关的 HOMA-IR 参数，而蜂胶只影响氧化相关的参数。
Undoubtedly, there is a gap in the literature data considering the evaluation of the bee products’ potential in human population. This results from the fact that bee products are characterized by very complicated biological matrix. Moreover, their composition varies depending on many factors, which are sometimes very difficult to control, that is, temperature. In fact, the chemical composition of each sample should be tested before being included in an in vivostudy. This is very tedious, time-consuming, and troublesome. Therefore, researchers today are more likely to use the commercially available equivalents, for example, CAPFE or pinocembrin. This is not exactly a good direction since we should remember that the complex products may exert synergistic effects. For example, Almaraz-Abarca et al.  evaluated the potential of bee pollen and chosen phenolic compounds in the inhibition of lipid peroxidation in microsomal preparations of mouse liver and showed much higher effectiveness of bee pollen (about 5.5 times as high as quercetin alone, 3 times as its glucoside (quercitrin) alone, and 2.4 times as caffeic acid alone).
毫无疑问，考虑到蜜蜂产品在人类种群中的潜力的评估，文献数据是有差距的。这是因为蜂产品是非常复杂的生物拥有属性。此外，它们的组成因许多因素而异，这些因素有时很难控制，即温度。事实上，在进行体内研究之前，每个样本的化学成份都应该进行测试。这是非常乏味、耗时和麻烦的。因此，今天的研究人员更倾向于使用商业上可以买到的替代品，例如 CAPFE 或 pinocemrin。这确实不是一个好的方向，因为我们应该记住，复杂的产品可能发挥协同作用。例如，Almaraz-Abarca 等人评估了蜂花粉和选择的酚类化合物在小鼠肝微粒体制剂中抑制脂质过氧化的潜力，结果显示蜂花粉的有效性要高得多(大约是单独槲皮素的5.5倍，单独是其葡萄糖苷(槲皮苷)的3倍，单独是咖啡酸的2.4倍)。
The aforementioned in vitro and animal studies seem to confirm the usefulness of using bee products (propolis, bee pollen, and royal jelly) as natural agents capable of counteracting the effects of oxidative stress underlying the pathogenesis of numerous diseases or disorders, such as neurodegenerative disorders, cancer, diabetes, and atherosclerosis, as well as negative effects of different harmful factors and drugs (e.g., cytostatic agents). However, studies on their role in humans are very limited, and the existing ones have aimed mostly at evaluating the effect of the supplementation of commercially available extracts of propolis or royal jelly in healthy people or type 2 diabetes. Unfortunately, in the available literature, there is a lack of studies considering this issue in the context of neurodegenerative disorders or cancers, although promising results were obtained in animal studies. This may result from the fact that particular samples of bee products may have different compositions, so it is difficult to draw a general conclusion concerning their potential therapeutic application without a detailed chemical analysis.
In conclusion, future studies concerning the question if bee products could be a promising adjuvant in the therapy of oxidative stress-related disorders or diseases in human seem to be advisable.