Mechanism Of ZaoZhuYinChen Decoction Improve Non-Alcoholic Steatohepatitis In Rats Based On PINK1-Parkin Mediated Mitophagy

Objective:To observe the effect of ZaoZhuYinChen Decoction(ZZYCF)improve steatosis and inflammation in NASH induced by high-fat diet,to explore the effect of ZZYCF on mitophagy in NASH rats,and to verify the changes of PINK 1/Parkin signal pathway.Methods:After 1 week of adaptive feeding,28 male Sprague-Dawley rats were randomly divided into high fat diet model group and normal diet control group(group C,n=7).The rats in the high-fat diet group were fed with high-fat diet for 16 weeks.From the 9th week,the rats in the high-fat group were randomly divided into the following three groups(n=7):model group(M group),ZaoZhuYinChen Decoction group(ZZYCF group)and pioglitazone group(PH group).At the end of the 16th week,the levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),liver triglyceride(TG),total cholesterol(T-CHO)and low-density lipoprotein(LDL),were measured,liver histology was observed by HE staining,fat accumulation was observed by oil red O staining,and the expression level of MDA was detected.The expression levels of PINK1,Parkin,p62 and LC3 were detected by Western blotting,and the hepatocyte mitophagy was located by immunofluorescence staining,and the data were statistically analyzed.Results:Compared with group C,the levels of fat weight,fat-body weight ratio,liver wet weight and liver index in group M were significantly higher than those in group C,with diffuse steatosis and swelling of hepatocytes,necrosis,nuclear fragmentation or dissolution of some hepatocytes with inflammation,and serum ALT(P<0.001),AST(P<0.001)levels significantly increased.Liver fat accumulation was increased,triglyceride(P<0.001),low density lipoprotein(P<0.001)and total cholesterol(P<0.001)were significantly increased,oxidative stress injury was aggravated and liver MDA level was increased(P<0.001).The abundance of PINK1,Parkin and LC3 significantly decreased,the expression of p62 protein increased,the co-localization of LC3,PINK1 and COX Ⅳ decreased,and the level of mitophagy was inhibited.After intervention with ZaoZhuYinChen Decoction,the levels of fat weight,fat/body weight ratio,liver wet weight and liver index were significantly decreased,hepatocyte balloon degeneration and inflammatory infiltration area were significantly reduced,serum ALT and AST levels were significantly decreased,fat deposition was alleviated,triglyceride(P<0.001),low density lipoprotein(LDL)and total cholesterol(P<0.001)were significantly lower than those in group M,oxidative stress injury was improved and liver MDA level was decreased.The expression levels of PINK1,Parkin and LC3 in liver mitochondria significantly increased,and the accumulation of p62 significantly decreased.The co-localization of LC3,PINK1 and COX Ⅳ in group M was more than that in group M.Conclusion:ZZYCF can improve the state of steatosis and inflammatory injury in non-alcoholic steatohepatitis.And maintain the normal homeostasis of mitochondria,alleviate the disorder of liver steatosis and lipid metabolism by activating mitophagy mediated by PINK1/Parkin signal pathway.