Study On The Mechanism Of Qutan Huoxue Recipe Regulating Pink1/Parkin Signaling Pathway To Induce Autophagy Of NASH Rat Hepatocytes

Objective:To investigate the mechanism of Qutan Huoxue Recipe to Reli ef NASH rats by regulating PINK1/Parkin signaling pathway to induce h epatocyte autophagy.Methods:1.Forty sexually clean SD rats were randomly divided into two groups:control group(N = eight)and Nash Group(N = thirty-two).The control group was given ordinary feed,and the NASH Group was given high fat diet.after eight weeks to confirm the success of mold,divided the NASH group to model group,low-dose Qutan Huoxue Recipe(6.1 g/kg),middle-dose group(12.2 g/kg)and high-dose group Qutan Huoxue Recipe(24.3 g/kg)randomly.The Rats in the model group and Qutan Huoxue Recipe group were given high-fat diet,and the rats in each Qutan Huoxue Recipe group were continuously given Qutan Huoxue Recipe for 4 weeks,and the rats were killed at the end of the twelve week.2.Weigh and record the body mass and liver mass of rats;HE staining and oil red O staining to observe the lipid deposition of hepatocytes and liver inflammation;electron microscopy to observe the number of lipid droplets and autophagic vesicles in liver tissue;take the serum to determine the content of ALT,AST,TG and TC to observe the liver injury of NASH rats;3.Immunohistochemistry and Western blot experiments were performed to observe the expression of PINK1,Parkin and SQSTM1 in rat liver.Results:1.Compared with the control group,the rats in the model group had increased body weight,sluggish movement,poor spirit and greasy fur.After the administration of Qutan Huoxue Recipe,the general condition of the rats improved,the body weight and liver weight decreased and the activity increased.2.The liver function biochemical indexes and blood lipids of the rats in the model group were higher than those in the control group,and ALT,AST,TG and TC decreased after the administration of Qutan Huoxue Recipe.3.HE staining and oil red O staining of liver tissue showed that compared with the control group,the liver tissue structure of the model group rats was incomplete,with disordered cell arrangement and a large number of fat vacuoles inside.Inflammatory cell infiltration was observed locally.After treatment with Qutan Huoxue Recipe,the liver tissue structure damage and cell arrangement disorder were alleviated compared to the model group,while fat vacuoles were reduced compared to the model group.Among them,the high-dose group showed the most significant recovery of liver tissue structure and reduction of fat vacuoles;A large number of red lipid droplets were observed in the liver tissue of the model group,with visible vacuoles and swelling of some liver cells.After treatment with Qutan Huoxue Recipe,the number of lipid droplets decreased,the cell structure was clear,and no obvious vacuoles were observed.The high-dose group had the best improvement effect.4.The number of lipid droplets and autophagic vesicles in rat liver tissues were detected by transmission electron microscopy,and it was observed that no obvious autophagic vesicles were seen in the control group and the model group,while a large number of lipid droplets were seen in the model group,and the number of autophagic vesicles increased and the number of lipid droplets decreased and their size decreased in the Qutan Huoxue Recipe group.5.The immunohistochemical results that the rats in the control group had no positive expression of SQSTM1 and lower positive expression of PINK1 and Parkin in the liver;compared with the control group,the model group had no positive expression of PINK1 and Parkin and higher expression of SQSTM1;compared with the model group,the rats in the Qutan Huoxue Recipe group had increased positive expression of PINK1 and Parkin in the liver and decreased expression of SQSTM1 expression was decreased.6.The Western blot experiment showed that the expression of autophagy proteins PINK1,Parkin and SQSTM1 was reduced in the model group,and the expression of PINK1,Parkin and SQSTM1 was increased after treatment with Expectorant and Qutan Huoxue Recipe.Conclusion: Qutan Huoxue Recipe may play a role in the treatment of NASH by inducing hepatocyte autophagy through regulating the PINK1/Parkin signaling pathway and reducing hepatocyte inflammation and liver tissue lipid deposition in NASH rats.