Based On AMPK/SIRT1/UCP2 Pathway To Explore The Effect Of Shugan Huazhi Capsule On Liver Oxidative Stress In NAFLD Rats

Purpose:In this experiment,a non-alcoholic fatty liver disease(NAFLD)animal model was established by establishing high-fat diet-fed rats,and Shuganhuazhi capsules were used for drug treatment to observe the liver tissue morphology and lipids of Shuganhuazhi capsules on NAFLD rats.The effects of Shuganhuazhi Capsule on NAFLD rats and its possible mechanism were investigated.Material and method:Thirty male Sprague-Dawley rats were fed one week and randomly assigned to three groups according to the random number table method.They were normal group,model group,and traditional Chinese medicine group,10 in each group.Except for the normal group,the other two groups were fed with high-fat diet.Eight weeks later,gavage was started.The Chinese medicine group was given gavage of traditional Chinese medicine.Normal and model groups were given intragastric gavage as a control.Rats in each group were sacrificed after 4 weeks of treatment,and serum and liver tissues were collected.HE staining was used to observe the pathological changes of rat liver tissue.The serum of triglycerides(TG),total cholesterol(TC)and alanine aminotransferase were detected by automatic biochemical analyzer.(ALT),aspartate aminotransferase(AST),high-density lipoprotein-cholesterol(HDL-C),and low-density lipoprotein-cholesterol(LDL-C))Level;Enzyme-linked immunosorbent assay(ELISA)was used to determine the oxidative stress factors superoxide dismutase(SOD),methane dicarboxylic aldehyde(MDA),and glutathione peroxidase in liver tissues.,GSH-Px)level;PCR was used to detect AMP-activated protein kinase(AMPK),silent information regulator(SIRT1)and uncoupling protein in liver tissue of each group.Uncoupling protein2,UCP2)gene expression;The protein levels of AMPK,SIRT1 and UCP2 in liver tissues were detected by Western Blot.Results:1.Liver histopathology resultsObservation of HE staining: In the normal group,hepatocytes were blue-stained,the cytoplasm was uniformly red-stained,the hepatic lobules were clear,the hepatocytes were normal in structure,uniform in size,and the nucleus was normal.There was no lipid droplet and vacuole in the center,and no obvious inflammatory cells were observed.Infiltration and steatosis,necrosis.In the model group,the structure of hepatic lobule was disordered and hepatocytes were obviously swollen.There were a large number of lipid droplets of different sizes in the cytoplasm.Inflammatory cell infiltration was observed,showing moderate and severe diffuse bullous steatosis.Compared with the model group,the hepatic tissue of the Chinese medicine group had improved in the aspects of hepatic lobule structure,outline,hepatocyte morphology,and inflammatory cell infiltration.2.Biochemical Indicators2.1 Serum TG,TC,HDL-C,LDL-CThe serum levels of TG,TC,and LDL-C in the model group were significantly higher than those in the normal group(P<0.05),while the levels of HDL-C were significantly lower(P<0.05).Compared with the model group,the levels of serum TG,TC,and LDL-C in the traditional Chinese medicine group were decreased,the difference was statistically significant(P<0.05),while the level of HDL-C was increased(P<0.05).2.2Serum ALT,AST contentThe serum ALT and AST levels in the model group were significantly higher than those in the normal group(P<0.05).Compared with the model group,the contents of ALT and AST in the traditional Chinese medicine group were significantly decreased,and the difference was statistically significant(P<0.05).3.Expression of SOD,MDA and GSH-Px oxidative stress factors in liver tissueCompared with the normal group,the SOD and GSH-Px levels in the liver tissue of the model group were all decreased,the difference was statistically significant(P<0.05),but theMDA level in the rat was significantly increased,the difference was statistically significant(P<0.05);Compared with the model group,the SOD and GSH-Px levels in the liver tissue of the TCM group were significantly higher(P<0.05),while the MDA levels were lower,the difference was statistically significant(P<0.05).4.Liver tissue AMPK,SIRT1,UCP2 gene and protein expressionhe expression of AMPK and SIRT1 gene and protein in the model group were lower than those in the normal group,the difference was statistically significant(P<0.05),while the UCP2 gene and protein expression were increased,the difference was statistically significant(P<0.05);Compared with the model group,the mRNA and protein expression of AMPK and SIRT1 in the TCM group increased,the difference was statistically significant(P<0.05).The UCP2 gene and protein expression were down-regulated,and the difference was statistically significant(P<0.05).Conclusion:1.Shuganhuazhi Capsule can significantly improve the liver tissue morphology of nonalcoholic fatty liver disease model rats,and it is an effective prescription for the treatment of non-alcoholic fatty liver disease.2.Shuganhuazhi capsule can affect serum lipids(TG,TC,HDL-C,LDL-C)and improve liver function(ALT,AST),thereby improving NAFLD hepatic lipid deposition and reducing hepatic steatosis.3.Shuganhuazhi capsules can regulate the expression of related genes and proteins in the AMPK/SIRT1/UCP2 signaling pathway in liver tissue,thereby improving lipid metabolism,inhibiting hepatic steatosis,and adjusting oxidation and antioxidant imbalance.This part explains the mechanism of Shuganhuazhi Capsule in treatingNAFLD.