Experimental Study Of Protective Mechanism Of Silibinin By Wnt/beta-catenin Signaling Pathway On The Alcoholic Liver Fibrosis Model Rats

ObjectiveTo evaluate the efficacy of Silibinin capsules in the treatment of alcohol-induced liver fibrosis in rats and explore the underlying mechanism to prevent the liver damage in ALF from anti oxidative stress and improving liver function and reversing the degree of liver fibrosis and the influence of Wnt/β-catenin signaling pathway.Methods15 healthy male SD rats were used as normal control group, the rest of the rats were copied alcohol liver fibrosis model by ethanol mixture.The successful model of the rats were randomly divided into the model group、Silibinin Capsule high-dose group(75.6mg·kg-1), middle-dose group(37.8mg·kg-1) and low-dose groups(18.9mg·kg-1)、positive group(colchicine,0.27mg·kg-1),15 rats in each group.Successive administration for 4 weeks, after the last administration, separation of serum and taking the liver.Liver tissue morphology was measured by HE staining.Hepatic fibrosis was evaluated by Masson Trichrome staining. The levels of AST,ALT were determined by using spectrophotometry.The concentration serum HA、LN、Hyp levels and liver tissue PCⅢ、Ⅳ-C、SOD、MDA、TGF-β1、TIMP-1 levels were measured by ELISA.The α-SMA expression by immunohistochemical methods in the liver of each group.The expression quantity of liver of Wnt、β-catenin、MMP-7 protein by Western blotting.Results1.HE staining was observed that the structure of liver tissue in normal group was clear and the morphology of liver cells was normal.The normal structure of liver tissue of rats in model group were damaged, liver cell swelling, degeneration, necrosis, with a large stream of water degeneration and steatosis. Hepatic cords disorders; portal areaand central vein showed a large number of inflammatory cells infiltration, fibrous connective tissue hyperplasia, and extends to the inside lobule, area of the form is incomplete.Compared with the model group, the treatment group of pathology improved significantly: liver damage, liver cell edema and the degree of steatosis were relatively eased, portal area connective tissue hyperplasia was not obvious.2.Masson staining observation can be seen that the normal group only having a little collagen deposition in the liver tissue mainly located in the portal area. Collagen fibers in liver tissue of the model rats hyperplasia was obviously. There were a lot of collagen deposition in portal area 、 central vein and the liver parenchyma, into the hepatic lobule forming incomplete interval, showed the formation of liver fibrosis.Compared with the model group, the collagen deposition in the treatment group was significantly decreased.3.Compared with the normal group,the levels of serum AST、ALT、HA、LN、Hyp were significantly higher(P<0.01) in model group, the relative expression quantity of Wnt 、 β-catenin 、 MMP-7 protein significantly increased in model group(P<0.01).Compared with model group,the levels of serum AST、ALT、HA、LN、Hyp were significantly decreased(P<0.05 or P<0.01)and the relative expression quantity of Wnt、β-catenin、MMP-7 protein significantly reduced(P<0.05 or P<0.01) in drug groups.4.Compared with the normal group,the liver tissue PCⅢ、Ⅳ-C、MDA、TGF-β1、TIMP-1levels were significantly increased(P<0.01),SOD levels dropped significantly(P<0.01)in model group.Compared with the model group,the levels of PCⅢ 、 Ⅳ-C 、 MDA 、 TGF-β1 、 TIMP-1decreased significantly( P<0.05 or P<0.01), the levels of SOD increased significantly in drug groups(P<0.01).5.Compared with the normal group, the expression of α-SMA significantly increased(P<0.01)in model group;compared with the model group,α-SMA expression was significantly decreased in drug groups(P<0.01).Conclusion1.Silibinin capsule can effectively reduce the content of serum AST, ALT, HA, LN,Hyp and improve liver function of ALF;at the same time can reduce the pathological changes in liver tissues of ALF,and then,prevent and treat the liver damage either or organic in ALF.2.Silibinin capsule can effectively reduce the content of liver tissue MDA and at the same time increase the activity of SOD.The results show that Silibinin capsule has the protection of liver damage in ALF through antioxygenation.3.Silibinin capsule can significantly reduce the content of liver tissue PCⅢ、Ⅳ-C、TGF-β1 、 TIMP-1 、 the expression of α-SMA and decreased the relative expression quantity of Wnt 、 β-catenin 、 MMP-7 protein.The mechanism of prevention and treatment of liver damage in ALF with Silibinin capsule might be related to the regulation of Wnt/β-catenin signal pathway.