白藜芦醇对蛋氨酸胆碱缺乏培养基诱导肝细胞损伤的保护作用

PROTECTION OF RESVERATROL ON HEPATOCYTES AGAINST METHIONINE-CHOLINE-DEFICIENCY-INDUCED INJURIES

目的研究白藜芦醇对蛋氨酸胆碱缺乏(methionine-choline-deficient,MCD)培养基诱导的小鼠正常肝细胞(alpha monse liver 12,AML12)损伤的保护作用。方法体外培养小鼠正常肝细胞AML12,建立蛋氨酸胆碱缺乏培养基诱导的肝细胞损伤模型,用不同浓度白藜芦醇(25、50、100μmol/L)进行干预24h,检测细胞的增值活性,以及细胞培养基上清液中谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱基酶(LDH)活性和细胞中丙二醛(MDA)水平、甘油三酯(TG)含量,检测细胞中炎性细胞因子IL-1β、IL-6、TNF-α水平及其m RNA水平,并检测微管相关蛋白1轻链3(MAP1-LC3)蛋白的表达。结果与MCD损伤组相比,各白藜芦醇干预组细胞活力明显增强,白藜芦醇可以显著抑制MCD导致的肝细胞ALT,AST,LDH的释放,降低肝细胞内MDA水平和TG含量,降低炎性细胞因子水平,上调MAP1-LC3的表达。结论白藜芦醇对MCD诱导的的小鼠肝AML12细胞损伤有明显的保护作用,自噬可能在其中起到一定的作用。

Objective To investigate the effects of resveratrol on methionine-choline-deficiency-induced injuries in alpha mouse liver 12（ AML12） hepatocytes. Methods The mouse liver cell line AML12 was cultured and the methionine-choline-deficiency-induced injury model was set up. Cell viability, alanine transaminase（ALT）, aspartate transaminase（AST） and lactate dehydrogenase（ LDH） activities in culture supernatants and contents of triglyceride（TG） and malondialdehyde（MDA）in cell lysates were measured in hepatocytes incubated with resveratrol（25, 50, 100 μmol/l） for 24 h. The contents of IL-1β, IL-6 and TNF-αwere detected by ELISA methods, and the mRNA levels were determined by RT-PCR. Expression of microtubule-associated protein 1 light chain 3（MAP1-LC3） was measured by Western blot method. Results Compared with the methionine-choline-deficiency-induced injury group, resveratrol treatment increased the survival of hepatocytes, lowered activities of ALT, AST and LDH in culture supernatants, reduced the contents of MDA and TG and the levels of inflammatory factors IL-1β, IL-6 and TNF-α, increased MAP1-LC3 expression. Conclusion Resveratrol can significantly protect AML12 hepatocytes against the damages induced by methionine-choline-deficiency, and the underlying mechanism is possibly associated with the autophagy induced by resveratrol.