Study On The Mechanism Of Apigenin Against Nonalcoholic Fatty Liver Disease

The incidence of nonalcoholic fatty liver disease?NAFLD?is increasing year by year due to changes in people's living habits and dietary structure.NAFLD is a clinicopathological syndrome,which is characterized by excessive lipid deposition and steatosis in hepatocytes,besides liver injury caused by alcohol and other definite factors.Liver fibrosis and cirrhosis can occur in patients with severe NAFLD,which seriously endangers life and health.So far,NAFLD has not approved therapeutic drugs.In this study,NAFLD mice model was established by high fat diet and the classical model of nonalcoholic steatosis of L-02 cells was established by palmitic acid,To explore the mechanism of apigenin in anti-nonalcoholic fatty liver through in vivo and in vitro experiments.Objective Study on the anti-nonalcoholic fatty liver effect and mechanism of apigenin in vitro and in vivo.Methods Cell experiment:Recovery,digestion,passage and cryopreservation of human normal hepatocyte line L-02 cells by routine methods.Cells in logarithmic growth phase were selected for modeling and medication,lipid deposition was observed,triglyceride?TG?content was detected,and JAK2/STAT3-mediated signal pathway-related protein expression was detected by Western Blot.Animal experiment:Sixty healthy male C57 mice were randomly divided into normal group,model group and apigenin group with 20 mice in each group.The normal mice were fed with basic diet,and the model group and apigenin group were fed with high fat diet to establish the model of non-alcoholic fatty liver in mice.After 8 weeks of feeding,mice in each group were given corresponding drugs by gavage every day for 16 weeks.Apigenin group was given 200 mg·kg^-1·d^-11 0.1ml/10g apigenin by gavage,while normal group and model group were given the same dose of solvents by gavage.During the period of drug treatment,the general situation of mice was observed.After 16 weeks of successful modeling and medication,all mice fasted for 12 hours,then all mice were weighed,blood was collected from eyeballs and serum was separated by centrifuge.GLU,LDL,TG,AST,ALT,TC were detected by automatic biochemical analyzer;mice were killed and their livers were extracted.Calculate liver index;observe liver histopathological sections by HE staining and oil red O staining;detect the content of TG in liver tissue in some liver homogenates;detect the expression of JAK2/STAT3-mediated signaling pathway-related proteins by protein immunoblotting;detect the expression level of liver lipid metabolism-related and inflammation-related genes by reverse RT-PCR.Result?1?Compared with the normal group,oil red O staining showed that lipid aggregation was obvious in the model group,compared with the model group,lipid aggregation was improved in different concentration groups of apigenin,and decreased in the high concentration group;compared with the normal group,the cell TG content in the model group increased significantly?P<0.01?.Compared with the model group,the cell TG content in the apigenin high concentration group decreased significantly?P<0.01?;compared with the normal group,the expression ratios of P-JAK2/JAK2,P-STAT3/STAT3 in the model group were significantly higher?P<0.01?;compared with the model group,the expression ratios of P-JAK2/JAK2,P-STAT3/STAT3 in the high concentration apigenin group were significantly lower?P<0.01?.?2?Compared with the normal group,the liver index of mice in the model group increased significantly?P<0.01?,and the liver index of mice in the apigenin group decreased significantly?P<0.01?;compared with the normal group,the levels of serum biochemical indexes such as TG,TC,GLU and LDL in the model group increased significantly?P<0.05 or P<0.01?,Compared with the model group,the levels of serum biochemical indexes such as TG,TC,GLU and LDL in the apigenin group decreased significantly?P<0.05or P<0.01?;compared with the normal group,the levels of serum ALT and AST in the model group increased?P<0.05 or P<0.01?.Compared with the model group,the levels of serum ALT and AST in the apigenin group decreased significantly?P<0.05 or P<0.01?;compared with the normal group,the content of TG in liver tissue of model group was significantly higher?P<0.01?.Compared with model group,the content of TG in liver tissue of apigenin group was significantly lower?P<0.01?;Oil red O staining showed a large amount of lipid deposition in the liver tissue of the model group,and there was a significant improvement in the apigenin group.HE staining showed that the liver of the model group had obvious steatosis,and the apigenin group had a significant improvement;compared with the normal group,the expression ratio of P-JAK2/JAK2 and P-STAT3/STAT3 protein in the model group increased significantly?P<0.05?.Compared with the model group,the expression ratio of P-JAK2/JAK2 and P-STAT3/STAT3 protein in the apigenin group decreased significantly?P<0.05?;compared with the normal group,the expression of IL-6,TNF-?Fas and srebf-1 in the model group increased,with significant difference?P<0.05 or P<0.01?.Compared with the model group,the expression of IL-6,TNF-?,Fas and srebf-1 in the apigenin group decreased,with significant difference?P<0.05 or P<0.01?.Conclusion Apigenin inhibits nonalcoholic fatty liver disease by inhibiting JAK2/STAT3-mediated signaling pathway.