Effect Of Metformin On Acute Liver Injury Induced By Carbon Tetrachloride In Mice By Inhibiting PI3K-AKT-mTOR Signaling Pathway

Objective:Using the hepatotoxic drug carbon tetrachloride(CCL4)to construct the mouse model of acute liver injury,to investigate the effect of metformin on CCL4-induced acute liver injury in mice by inhibiting the PI3K-AKT-m TOR signaling pathway,and to provide a new idea for clinical targeted treatment of acute liver injury.MethodsSelected 18 only six to eight weeks of healthy male C57 BL / 6 mice random grouping: normal control group,CCL4 model group and CCL4 + metformin intervention group(n = 6),model group and intervention group by intraperitoneal injection of 40%CCL4(5 m L/kg)acute liver injury in mice model was constructed,and the normal control group given isotonic saline,intraperitoneal injection of intervention group metformin before building(soluble in saline solution,according to the weight)400mg/kg doses in mice preconditioned 6 h,the rest of the two groups of isotonic saline.After 18 h,the anesthetized mice were selected for eyeball and blood collection to detect ALT and AST.The ratio of liver weight to body weight was calculated.Paraffin embedded liver tissue was used to prepare sections,and HE staining was used to observe the pathological changes of mouse liver.The expression levels of inflammatory cytokines(IL-6,TNF-?,IL-10)and their gene m RNA in liver tissues of mice were detected by ELISA and RT-q PCR.The expression levels of inflammatory cytokines(IL-6,TNF-?,IL-10)and their gene m RNA in liver tissues of mice were detected by ELISA and RT-q PCR.The phosphorylation levels of PI3 K,AKT,and m TOR proteins,as well as the expression levels of the transcription factor ROR-? t and Foxp3 proteins in Th17 cells and Treg cells in mouse liver tissues were detected by Western-blot.One-way anova and LSD-t were used for statistical analysis.Results:Compared with normal control group,CCL4 model group serum transaminase(ALT,AST),liver weight and liver inflammation factors(IL-6,TNF-?,IL-10)expression level was significantly increased(P < 0.05);Balloon degeneration and necrosis of numerous hepatocytes and inflammatory infiltration of liver tissue;Phosphorylation levels of PI3 K,AKT,m TOR,ROR-?t,and Foxp3 were all increased in liver tissues(P<0.05).In the CCL4 model + metformin intervention group,compared with the CCL4 model group,serum transaminase and liver weight ratio decreased significantly.The denatured and necrotic areas in the liver were significantly reduced,and the infiltration of inflammatory cells was also significantly reduced.The level of inflammatory cytokines(TNF-?,IL-6)in liver homogenate decreased,while the level of inflammatory cytokines(IL-10)increased.In liver tissues,the phosphorylation levels of PI3 K,AKT,and m TOR,as well as the expression of ROR-?t protein all decreased,and the expression of Foxp3 increased significantly(P<0.05).Conclusion:1.40%CCL4(5 m L/kg)was intraperitoneally injected into mice for 18 hours to establish a stable mouse model of acute liver injury.2.Metformin can regulate the inflammatory response by inhibiting the PI3KAKT-m TOR signaling pathway,so as to reduce the CCL4 induced acute liver inju ry in mice,providing a new idea for the clinical treatment of drug-induced liver injury.