Effects Of Liraglutide On The Expression Of LC3??p62 In NAFLD Cell Induced By High-fat In Vitro

Objectives: The aim of this study was to investigate the effects of glucagon-like peptide 1(GLP-1) receptor agonist Liraglutide on the the expression of cell autophagy related indicators LC3? and p62, induced by high-fat in L02 cells and its possible mechanisms. Expect to provide a new theoretical basis for the application of GLP-1 in the treatment of nonalcoholic fatty liver disease(NAFLD).Methods: L02 cells were cultured in vitro with DMEM medium containing 10% fetal bovine serum(FBS). Oleic acid, palmitic acid(2:1) to induce hepatic steatosis, nonalcoholic fatty liver disease(NAFLD) model is established. The experiment was grouped as follows: 1) control group; 2) NAFLD;3) high-fat + Liraglutide groups: cultures of fatty medium, 10 nmol/L join Liraglutide;4) NAFLD cell+ Liraglutide groups+normal cultures: medium FBS proportion 10%, and Liraglutide concentration 10nmol/L;5) normal group+ Liraglutide: normal medium and Liraglutide concentration 10nmol/L; Cells of all groups were intervened for 48 h, respectively. After intervention, the levels of Triglyceride(TG), Alanine Aminotransferase(ALT), were measured. Lipid droplets in the cells were observed through oil red staining. The mRNA expressions of cell autophagy related indicators LC3? and p62 were detected by RT-PCR.The protein levels of LC3? and p62 were detected by Western blot. Using the method of TUNNEL testing the change of cell apoptotic. The data were presented as SPSS 23.0 software for statistical analysis, measurement data use mean ± standard deviation. Experimental data are normal distribution test and f test, if conform to the normal distribution using the Factor analysis of variance(one- way ANOVA),to does not conform to the normal distribution of the K- W rank-sum test.The difference was statistically significant with P<0.05.Results:1 The liraglutide of normal L02 cells,TG, ALT level,LC3??p62 mRNA and protein expression had no obvious effect(P > 0.05).2 Compared with normal control group, intracellular lipid droplets of NAFLD group was obviously increased, TG?ALT?MDA levels, LC3? mRNA and protein expression are reduced,p62 mRNA and protein expression are increased, apoptosis index increased, the differences were significant(P< 0.05).3 Compared with NAFLD cell model group, NAFLD cell model + liraglutide + high-fat culture, NAFLD cell model+ the liraglutide + normal culture lipid droplets in the cell culture group was obviously reduced, TG, ALT,MDA levels drop, LC3??p62 mRNA and protein expression decreased, apoptosis index fell, the differences were significant(P<0.05).4 Compared with NAFLD cell model + the liraglutide +the normal culture group, NAFLD cell model + the liraglutide + the high-fat culture group in lipid droplets decrease rate is low, TG, ALT,MDA levels drop, LC3??p62 mRNA and protein expression and apoptosis index levels change is low, differences were significant(P<0.05).Conclusion:1 Established the model of nonalcoholic fatty liver disease in vitro cell;2 The glp-1 receptor agonist liraglutide can reduce liver lipid deposition in cells, improve the function of liver cells.3 Liraglutide can increase L02 cell autophagy related protein expression levels, improve cell lipid metabolism, slow down the development of NAFLD.