余甘子多酚对体外酒精性肝损伤的保护作用

Hepatoprotective Effect of Polyphenols of Phyllanthus emblica L. against Alcohol-Induced Oxidative Damage in Vitro

目的:研究余甘子多酚(PPE)对酒精性肝细胞的保护作用,以及核因子相关因子2(Nrf2)/血红素氧合酶-1(HO-1)信号通路在其中的调控作用。方法:建立BRL-3A细胞酒精性损伤模型,用试剂盒检测PPE干预前、后细胞培养上清液中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、谷胱甘肽(GSH)含量;用Q-PCR法检测Nrf2和HO-1的mRNA表达水平,Western印迹法检测细胞蛋白表达。结果:余甘子多酚可降低MDA、ALT、AST含量,提高SOD和GSH水平;降低Nrf2的mRNA水平和蛋白质含量,阻止Nrf2的过度表达;上调HO-1的m RNA水平和蛋白质含量,促进细胞抗氧化酶基因HO-1的表达,增强细胞自身抗氧化能力,抑制酒精性肝细胞氧化损伤。结论:余甘子多酚通过调控Nrf2/HO-1信号通路抑制酒精性肝细胞损伤,其主要作用成分为没食子酸、柯里拉京和鞣花酸。

Objective: This study was aimed to investigate the protective effects of polyphenols from Phyllanthus emblica L.(PPE) against ethanol-induced oxidative damage in rats liver cell, and further reveal the molecular mechanisms by examining the Nrf2/HO-1 pathway. Methods: BRL-3 A mice were used to evaluate the hepatoprotective effect of PPE on liver injury induced by ethanol exposure. The biochemical parameters(MDA, SOD, ALT, AST and GSH) were measured using commercial kits. HO-1 mRNA level was determined by Q-PCR. Protein levels were measured by western blot. Results: A decreased amount of MDA, ALT and AST release were observed, whereas PPE pretreatment significantly increased the leakage of SOD and GSH in BRL-3A cells. Furthermore, PPE pretreatment apparently up-regulated HO-1 expression, and prevented the overexpression of Nrf2 expressions. Conclusion: These results demonstrate that PPE-mediated up-regulation of HO-1 via Nrf-2 signaling pathway may provide a pivotal mechanism for its hepatoprotective activity.The phenolic compounds including gallic acid, corilagin and ellagic acid present in PPE might be playing a major role against alcohol-induced oxidative damage in rat cells.