The Endoplasmic Reticulum Stress Effect Of Liraglutide Treating Nonalcoholic Fatty Liver Cell Model Induced By In Vitro

Objectives: By observing the glucagon like peptide 1(glp-1) receptor agonist Liraglutide intervention treating nonalcoholic fatty liver induced by in vitro cell model of detection of endoplasmic reticulum stress related target GRP78 protein expression, ATF6 nucleoproteid level and the change of caspase12, explore the mechanism and effect for liraglutide treating NAFLD.Methods:1 Cell model of non- alcoholic fatty liver induced in vitro : in vitro using 10% fetal bovine serum(FBS) DMEM to culture normal human liver cell L02 cells, adding oleic acid, palmitic acid(2:1) to induce hepatic steatosis, to establish the model of nonalcoholic fatty liver disease(NAFLD).2 Cell grouping and intervention: 1) normal control group; 2) NAFLD model group; 3)NAFLD model group and liraglutide treatment group and high fat medium group: high fat medium was added to 10 nmol / L liraglutide;4) NAFLD model group and liraglutide treatment group and normal nutrient solution group: the normal nutrient liquid was added to 10 nmol / L liraglutide; 5) the normal group + liraglutide: normal culture fluid was added to 10 nmol / L liraglutide; each group was treated for 48 h and collected. To detect the content of fat and metabolism in cells.3 Application of full automatic biochemical analyzer was used to determine the content of triglyceride(TG)in cell and Alanine aminotransferase in supernatant(Alanine Aminotransferase, ALT), staining was used to measure intracellular lipid droplets by oil red "O".4 The detection of oxidative stress related indicators MDA and SOD levels.5 Western blotting and qRT-PCR were used to detect the expression of GRP78 protein in the endoplasmic reticulum stress related indicators and the changes of nucleoproteid of ATF6 in the lower reaches and the changes of apoptosis index caspase12.6 TUNNEL analysis was used to detect the changes of cell apoptosis.7 Data analysis: The data were presented as SPSS 23 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.Result:1 The liraglutide of normal L02 cells, GRP78, ATF6, TG, ALT level caspase12 mRNA and protein expression had no obvious effect(P > 0.05).2 Compared with normal control group, intracellular lipid droplets NAFLD group was obviously increased, TG、ALT levels, GRP78, ATF6, caspase12 m RNA 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 lipid droplets in the cell culture group was obviously reduced, TG, ALT levels drop, GRP78, ATF6, caspase12 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 comparison, NAFLD cell model + tthe liraglutide + the high-fat culture group in lipid droplets decrease rate is low, TG, ALT levels drop, GRP78, ATF6, caspase12 mRNA and protein expression and apoptosis index levels drop is low, differences were significant(P < 0.05).Conclusion:.1 In high fat environment, liver cell apoptosis increases, while the liraglutide can against liver cell apoptosis.2 The glp-1 receptor agonist liraglutide can reduce liver lipid deposition in cells, improve the function of liver cells.3 The glp-1 receptor agonist liraglutide may cut GRP78, through ATF6 nucleoprotein express to reduce endoplasmic reticulum stress, the steady state of cell and thus reduce the expression of caspase- 12 inhibiting liver cell apoptosis, treatment of nonalcoholic fatty liver disease.