二甲双胍对非酒精性脂肪性肝病细胞模型内质网应激与自噬的影响

目的 观察二甲双胍对游离脂肪酸(FFA)诱导的非酒精性脂肪性肝病(NAFLD)HepG2细胞模型内质网应激(ERS)及自噬的影响.方法 2020年7—11月于武汉大学人民医院消化系统疾病湖北省重点实验室进行实验.分别用0(空白对照)、10、20、40、80 mmol/L的二甲双胍处理HepG2细胞24 h,评估二甲双胍...

Effects of metformin on endoplasmic reticulum stress and autophagy in a cell model of non-alcoholic fatty liver disease

Objective To observe the effect of metformin on the endoplasmic reticulum stress(ERS)and autophagy in a HepG2 cell model of non-alcoholic fatty liver disease(NAFLD)induced by free fatty acid(FFA).Methods From July 2020 to November 2020,the experiment was conducted at the Hubei Key Laboratory of Digestive System Diseases,Renmin Hospital of Wuhan University.HepG2 cells were treated with metformin at concentrations of 0,10,20,40,and 80 mM for 24 hours,and the cell counting kit was used to evaluate the effect of metformin on cell viability;the concentration within 10 mmol/L was selected for subsequent experiments.The cellswere divided into four groups:control group(BSA group),model group(FFA group),metformin low-concentration group(Met L group)and metformin high-concentration group(Met H group).HepG2 cells were treated with FFA for 24 hours to build NAFLD cell models and then metformin were added to interfere with the process.Western blot was used to detect the expression levels of ERS-related proteins phosphorylated PERK(p-PERK),ATF4 and autophagy-related proteins p62 and LC3 in HepG2 cells,and the expression of ATF4 mRNA was detected by real-time fluorescent quantitative PCR(qRT-PCR).Results Metformin can reduce the viability of HepG2 cells in a dose-dependent manner(F/P=1759.000/0.000).Compared with the control group,the expression levels of p-PERK,p62,ATF4 protein and mRNA in the model group increased(t/P=3.273/0.029,16.190/0.000,47.290/0.000,13.730/0.000),and the ratio of LC3Ⅱ/Ⅰdecreased(t/P=17.980/0.000);compared with the model group,the expression levels of p-PERK,p62,ATF4 protein and mRNA in the metformin low-concentration group and high-concentration group decreased(F/P=70.310/0.000,106.700/0.000,995.600/0.000,66.960/0.000),and the ratio of LC3Ⅱ/Ⅰincreased(F/P=166.400/0.000).Conclusion Metformin can reduce the level of ERS and increase the level of autophagy in NAFLD cell models induced by free fatty acids.