Discussing The Molecular Mechanism About The Intervention By Clearing Heat、Expelling Damp And Promoting Blood Circulation For Hepatic Fibrosis Based On Wnt/β-catenin Signaling

Introduction:Liver fibrosis is a reversible wound healing response which refers to a variety of pathogenic factors that leading to hepatocytes inflammation and necrosis,the imbalance between degradation and deposition of collagen and other extracellular matrix (ECM),leading to abnormal proliferation of liver connective tissue in the pathological state.At the progress of hepatic fibrosis,the activation of hepatic stellate cells secrete and contribute to the excessive deposition of I type collagen and other extracellular matrix.The emerging research of Wnt/p-catenin signaling shows that there is a certain relationship among the Wnt/β-catenin pathway and activation of hepatic stellate cells and liver fibrosis.The Chinese Medicine etiology and pathogenesis of liver fibrosis are complicated and changeful,the key pathogenesis is the damp-heat exist,so that the damp-heat and the blood stasis fit together with each other,the liver collaterals is damaged.Our previous clinical research shows that the molecular mechanism about the intervention for hepatic fibrosis of Modified Yinshao Powder which is based on clearing heat and expelling damp and promoting blood circulation,is effective for the treatment of chronic hepatitis and liver fibrosis,can alleviate and reverse the progression of hepatic fibrosis.So establishing the rules of treatment and exploring the mechanism about the intervention for hepatic fibrosis,can preferably provide theoretical basis for clinical treatment.Aim:Through observing the serum levels of ALT、AST、HA、IV type collagen、PIIINP、 LN of the rats,the liver tissues pathological changes,the expression lever of mRNA and protein of TGF-β、β-catenin,further discussed the molecular mechanism about the intervention of clearing heat and expelling damp and promoting blood circulation for hepatic fibrosis in rats,looking for new targets for TCM prevention and treatment of liver fibrosis and way.Methods:60 adult male Wistar rats divided randomly into the drug groups and the model group. We made models by giving subcutaneous injection of carbon tetrachloride,supplemented by a high-fat diet and 30% alcohol as the only drinking water,16 weeks.At the fifth week,begin to give intragastric administration with Traditional Chinese Medicine Yinshao powder and Colchicine to intervene the process of hepatic fibrosis,after 12 weeks sacrificed,detected the serum levels of ALT、AST、HA、IVtype collagen、PIIINP、 LN of the rats,observed the liver tissues pathological changes by HE staining and Masson staining,detect each rat liver tissue TGF-P and β-catenin expression situation by Immunohistochemical,detected the expression level of mRNA and protein of TGF-P and β-catenin by RT-PCR and Western Blot.Results:After copying the animal models and lavage intervention,the liver function and the four serum indices which reflecting the progress of liver fibrosis of the Chinese traditional medicine group is improved obviously;compared with colchicine group,serum ALT、AST,IV type collagen,HA,PⅢNP of the high dose group were significantly lower. We can observe that the structure of the rat liver tissues of the normal group are integrated,cell morphology are normal,liver cells is radial around the central vein,there are small amount of collagen fibers surrounding the cells;the structure of the liver tissues of the model group rats are destroyed,cells arrange are disordered,with a large number of vacuoles degeneration,a large amount of collagen surrounding the cells,there are also green stuff can be seen in the cells by HE staining and Masson staining.The damage degree of the structure of high dose group,and medium dose group is less,the lipid drpplets and the collagen fibers are produced,the lever is less than the model group and the Colchicine group.The Immunohistochemical results show that the positive expression of TGF-β、β-catenin of the model group are higher than drug group and colchicine group;the positive expression of TGF-β、β-catenin of the model group are higher than the chinese medicine groups and colchicine group、the positive expression of TGF-β、β-catenin of the colchicine group are higher than the high does group.Compared with the Colchicine group and model group,the expression lever of mRNA and protein of TGF-β and P-catenin in hepatic tissue of Traditional Chinese Medicine group is lower than models group and Colchicine group.Conclusion:The Modified Yinshao Powder which is based on clearing heat and expelling damp and promoting blood circulation,can alleviate the progression of hepatic fibrosis and protect the damaged liver function.The molecular mechanism about the intervention for hepatic fibrosis is related to inhibition or down-regulation of the expression of TGF-β and P-catenin that blocking the activation of TGF-β signaling pathway and Wnt/p-catenin signaling pathway.