This study aims to investigate the protective effects of royal jelly (RJ) and its functional components, major royal jelly proteins (MRJPs) and royal jelly lipids (RJL), against ethanol-induced oxidative damage in HepG2 cells, and to identify the core functional constituents responsible for the hepatoprotective activity of royal jelly. An oxidative damage model was established using 600 mmol/L ethanol. The effects of different concentrations (0.2, 0.4 mg/mL) of royal jelly and its extracts on cell viability, aspartate aminotransferase (AST), alanine aminotransferase (ALT), superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) levels in ethanol-exposed HepG2 cells were evaluated to systematically assess their regulatory roles in oxidative stress. Furthermore, RT-qPCR was employed to examine the influence of the bioactive hepatoprotective components of royal jelly on the mRNA expression levels of Nrf2, HO-1, TNF-α, IL-6, ADH, and PPARα, thereby elucidating the underlying molecular mechanisms of hepatoprotection. The results showed that the cell viability of the model group decreased to 45.61% after 24 hours of exposure to 600 mmol/L ethanol. At a treatment concentration of 0.4 mg/mL, RJ, MRJPs, and RJL increased the viability of ethanol-damaged HepG2 cells by 46.81%, 69.29%, and 93.57%, respectively, with the overall protective efficacy ranking as follows: RJL>MRJPs>RJ. Compared with the model group, pretreatment with RJ, MRJPs, and RJL dose-dependently inhibited the leakage of ALT and AST, and significantly elevated intracellular endogenous antioxidant levels across all treatment groups. Specifically, treatment with RJL at 0.2, 0.4 mg/mL reduced MDA levels by 43.02% and 67.07%, respectively, demonstrating a significantly stronger inhibitory effect on lipid peroxidation than RJ and MRJPs treatments at equivalent concentrations. Overall, RJL exhibited markedly superior performance in hepatocyte protection, antioxidant defense, and inhibition of lipid peroxidation compared to RJ and MRJPs, indicating that RJL may represent the principal hepatoprotective bioactive substance in royal jelly, with MRJPs playing a synergistic auxiliary role. Molecular mechanism studies revealed that RJL exerts its hepatoprotective effects primarily through direct modulation of the expression of genes associated with oxidative stress, inflammatory responses, and alcohol and lipid metabolism. This research provides a crucial theoretical basis for the development of functional hepatoprotective products and the high-value utilization of royal jelly.
[1] DEVARBHAVI H, ASRANI S K, ARAB J P, et al.Global burden of liver disease:2023 update[J].Journal of Hepatology, 2023, 79(2):516-537.
[2] 张效威, 刘宜杭, 张润东, 等.中医药治疗酒精性肝病的组方用药规律及作用机制研究[J].中药新药与临床药理, 2024, 35(8):1246-1254.
ZHANG X W, LIU Y H, ZHANG R D, et al.Analysis of prescription medication rules and mechanism of action of traditional Chinese medicine in the treatment of alcoholic liver disease[J].Traditional Chinese Drug Research and Clinical Pharmacology, 2024, 35(8):1246-1254.
[3] JAURIGUE M M, CAPPELL M S.Therapy for alcoholic liver disease[J].World Journal of Gastroenterology, 2014, 20(9):2143-2158.
[4] SONG X L, CUI W J, MENG F Y, et al.Glucopyranose from Pleurotus geesteranus prevent alcoholic liver diseases by regulating Nrf2/HO-1-TLR4/NF-κB signalling pathways and gut microbiota[J].Food & Function, 2022, 13(5):2441-2455.
[5] 褚佳豪, 居瑞军, 樊远红, 等.芦荟皮提取物对急性酒精性肝损伤的保护作用与机制[J].食品工业科技, 2023, 44(1):378-388.
CHU J H, JU R J, FAN Y H, et al.Protective effect and mechanism of Aloe bark extract on acute alcoholic liver injury[J].Science and Technology of Food Industry, 2023, 44(1):378-388.
[6] 阚默, 王继凤, 明思彤, 等.蜂王浆药理作用研究进展[J].特产研究, 2020, 42(6):55-59.
KAN M, WANG J F, MING S T, et al.Research progress of pharmacological effects of royal jelly[J].Special Wild Economic Animal and Plant Research, 2020, 42(6):55-59.
[7] CAIXETA D C, TEIXEIRA R R, PEIXOTO L G, et al.Adaptogenic potential of royal jelly in liver of rats exposed to chronic stress[J].PLoS One, 2018, 13(1):e0191889.
[8] 王金胜. 蜂王浆对CCl4导致小鼠肝损伤保护作用的研究[D].福州:福建农林大学, 2014.
WANG J S.Protective effects of royal jelly on carbon tetrachloride-induced liver injury in mice[D].Fuzhou:Fujian Agriculture and Forestry University, 2014.
[9] 冯彤彤, 张慧, 陈坤, 等.蜂王浆水溶蛋白对高糖诱导人脐静脉内皮细胞的保护作用[J].华西药学杂志, 2024, 39(4):392-394.
FENG T T, ZHANG H, CHEN K, et al.Protective effects of water-soluble royal jelly protein on high glucose induced human umbilical vein endothelial cells[J].West China Journal of Pharmaceutical Sciences, 2024, 39(4):392-394.
[10] 牛佳卉, 袁静, 张慧芳, 等.王浆酸对小鼠酒精性肝损伤的保护作用[J].食品工业科技, 2020, 41(3):291-296.
NIU J H, YUAN J, ZHANG H F, et al.Protective effect of royal jelly acid on the alcohol liver injury in mice[J].Science and Technology of Food Industry, 2020, 41(3):291-296.
[11] FOCAK M, SULJEVIC D.Ameliorative effects of Propolis and royal jelly against CCl(4)-induced hepatotoxicity and nephrotoxicity in wistar rats[J].Chemistry & Biodiversity, 2023, 20(1):e202200948.
[12] HU H, WEI Q H, SUN Z H, et al.Development of a freshness assay for royal jelly based on the temperature- and time-dependent changes of antimicrobial effectiveness and proteome dynamics of royal jelly proteins[J].Journal of Agricultural and Food Chemistry, 2021, 69(36):10731-10740.
[13] 陈伊凡. 蜂王浆脂肪酸的抗炎活性及其机制研究[D].杭州:浙江大学, 2019.
CHEN Y F.Anti-inflammatory effects and underlying mechanisms of royal jelly fatty acids[D].Hangzhou:Zhejiang University, 2019.
[14] TANG Z, ZHAN L, HE R R, et al.Hepatoprotective effect of tea composite solid beverage on alcohol-caused rat liver injury[J].Foods, 2023, 12(22):4126.
[15] RAMANATHAN A N K G, NAIR A J, SUGUNAN V S.A review on Royal Jelly proteins and peptides[J].Journal of Functional Foods, 2018, 44:255-264.
[16] 于心雨, 李珊珊, 陶凌晨, 等.蜂王浆中脂肪酸标志物检测方法的研究进展[J].食品工业科技, 2023, 44(3):499-507.
YU X Y, LI S S, TAO L C, et al.Recent advances in determination methods of fatty acid markers in royal jelly[J].Science and Technology of Food Industry, 2023, 44(3):499-507.
[17] YOU Y, MIN S, LEE Y H, et al.Hepatoprotective effect of 10% ethanolic extract from Curdrania tricuspidata leaves against ethanol-induced oxidative stress through suppression of CYP2E1[J].Food and Chemical Toxicology, 2017, 108:298-304.
[18] 兰天, 汪玲, 余贤娴, 等.蜂王浆及其提取物对失眠小鼠的睡眠改善作用[J].食品研究与开发, 2022, 43(20):83-90.
LAN T, WANG L, YU X X, et al.Effects of royal jelly and its extracts on sleep improvement in mice with insomnia[J].Food Research and Development, 2022, 43(20):83-90.
[19] 李喜艳, 王加启, 卜登攀, 等.多不饱和脂肪酸对细胞膜功能影响的研究进展[J].生物技术通报, 2009, 25(12):22-26.
LI X Y, WANG J Q, BU D P, et al.Advanced research of effect of polyunsaturated fatty acids on cell membrane function[J].Biotechnology Bulletin, 2009, 25(12):22-26.
[20] YUAN C Y, YANG S Z, GAO C M, et al.Biologically synthesized 10-Hydroxy-2-Decenoic acid activates cornified envelope formation via PPARα signaling, and regulates oxidative respiratory chain to improve mitochondrial function[J].Journal of Functional Foods, 2024, 122:106505.
[21] CHEN Y J, ZHANG X L, ZHENG Z H, et al.In silico prospecting of ADH activating peptides from Pacific oyster (Crassostrea gigas) and protective effect on ethanol-induced damage in HepG2 cells[J].Food Chemistry, 2025, 479:143777.
[22] TAKAHASHI K, SUGIYAMA T, TOKORO S, et al.Inhibitory effect of 10-hydroxydecanoic acid on lipopolysaccharide-induced nitric oxide production via translational downregulation of interferon regulatory factor-1 in RAW264 murine macrophages[J].Biomedical Research, 2013, 34(4):205-214.
[23] TANG M Y, WANG X Z, WANG S, et al.10-Hydroxy-2-decenoic acid attenuates nonalcoholic fatty liver disease by activating AMPK-α signaling pathway[J].Biochemical Pharmacology, 2025, 231:116648.
[24] FAN P, SHA F F, MA C, et al.10-hydroxydec-2-enoic acid reduces hydroxyl free radical-induced damage to vascular smooth muscle cells by rescuing protein and energy metabolism[J].Frontiers in Nutrition, 2022, 9:873892.
[25] 李宪秀, 何涛, 毛旸晨, 等.食叶草酵素对HepG2细胞酒精性损伤的保护作用及机制[J].食品科学, 2025, 46(14):179-186.
LI X X, HE T, MAO Y C, et al.Protective effect and mechanism of edible grass(Rumex patientia L.× Rumex tianschanicus A.LOS)jiaosu on alcohol-induced injury in HepG2 cells[J].Food Science, 2025, 46(14):179-186.
[26] 陈羽, 张清秀, 谷仕艳, 等.灯盏花素通过Nrf2信号通路改善HepG2细胞急性酒精性肝损伤[J].中南农业科技, 2024, 45(3):43-49;75.
CHEN Y, ZHANG Q X, GU S Y, et al.Ligustrazine improves acute alcoholic liver injury in HepG2 cells via the Nrf2 signaling pathway[J].South-Central Agricultural Science and Technology, 2024, 45(3):43-49;75.
[27] 林春燕, 赵甜甜, 刘俊, 等.鲟鱼籽酶解液对酒精诱导HepG2细胞氧化损伤的保护效果研究[J].食品与发酵工业, 2025, 51(17):93-99.
LIN C Y, ZHAO T T, LIU J, et al.Protective effect of sturgeon roe enzymatic hydrolysate powder on alcohol-induced oxidative damage of HepG2 cells[J].Food and Fermentation Industries, 2025, 51(17):93-99.
[28] ABOONABI A, SINGH I.Chemopreventive role of anthocyanins in atherosclerosis via activation of Nrf2-ARE as an indicator and modulator of redox[J].Biomedicine & Pharmacotherapy, 2015, 72:30-36.
