Experimental Study On The Effect Of Gansu Granule On Hepatic Fibrosis

Objective:Gansu granule is a Chinese patent medicine developed from Penthorum chinense Pursh.for the treatment of hepatitis.This study was to explore its role in the treatment of liver fibrosis and potential mechanism.Methods:1.Experiment of chemical hepatic fibrosis rats.Healthy SD rats were grouped into 6 groups randomly according to weight,which contained the control group,CCl₄induced model group,colchicine group（0.0001g/kg）,and Gansu Granule groups with different doses（16.7,8.4,4.2g/kg）.Animal models of liver fibrosis were constructed through intragastric administration of CCl₄,simultaneously with high fat and low protein diet Observe the general state of rats after given the corresponding medicine.Bloods were collected to detect the liver function by AST,ALT,TBIL,ALP ALB,and TP.Besides,the liver,spleen and thymus gland were dissected and weight.Then the liver was used to detect contents of HA,LN,MMP-1,and TIMP-1 by enzyme-linked immunoassay assay（ELISA）,pathological histology observation by HE staining and Masson staining.alpha-smooth muscle actin（α-SMA）and transforming growth factorβ₁（TGF-β₁）expression in the hepatic tissue was measured by immunohistochemical method.2.Experiment of immunologic hepatic fibrosis rats.Healthy SD rats were devided into 6 groups randomly according to weight,which contains control group,pig serum induced model group,colchicine group（0.0001g/kg）,and Gansu Granule groups with different doses（16.7,8.4,4.2g/kg）.Animal models of liver fibrosis were constructed through intraperitoneal injections of pig serum.Observe the general state of rats after given the corresponding medicine.Bloods were collected to detect the liver function by AST,ALT,TBIL,ALP ALB,and TP;besides,the liver,spleen and thymus gland were dissected and weight.Then the liver was used to detect contents of HA,LN,MMP-1,TIMP-1,PCⅢ,α-SMA,TGF-β₁ by enzyme-linked immunoassay assay（ELISA）,pathological histology observation by HE staining and Masson staining.3.Experiment of bile duct obstructive hepatic fibrosis rats.The bile duct obstructive hepatic fibrosis model was established by common bile duct ligation of rats.Then the model rats were divided into the model group,colchicine group（0.0001g/kg）and Gansu granule group（16.7g/kg）randomly according to weights.There was sham-operated group in which the rats were give sham operattion.After administered with drugs for 8 weeks,t liver function and liver histopathology were detected for all the groups.Results:1.Effects of Gansu granule on hepatic fibrosis model induced by CCl₄ in rats.Compared with the control group,ALT,AST,TBIL and ALP were significantly increased in the model group（P<0.01）,and the levels of HA,LN and TIMP-1 in the liver tissue were significantly increased（P<0.01）,with remarkably increase of liver,spleen coefficients and decrease of thymus gland coifficient.The liver of model rats were texture harden,extreme fibrosis and highly expression of TGF-β₁,andα-SMA in the liver.Compared with the model group,the serum levels of ALT,AST,ALP,TBIL and HA and TIMP-1 in the colchicine group and the Gansu Granule group were significantly decreased and the serum globulin was significantly increased（P<0.01）;The serum ALB level of the low dose group was significantly increased（P<0.05）;The degree of fibrosis in the treatment group was improved,and the expression of TGF-β₁andα-SMA in the liver tissue was weakened.2.Effects of Gansu Granule on immunological hepatic fibrosis rats.Compared with the control group,the levels of serum ALT,AST,ALP and TBIL were significantly increased in the model group,the ALB content was significantly decreased,the spleen coefficient was significantly increased,the thymus coefficient was significantly decreased,liver tissue HA,LN,The levels of TIMP-1,α-SMA and TGF-β₁ were significantly increased（P<0.05 or P<0.01）.The liver texture was hardened and pathological changes were observed.Pathological examination showed significant changes in fibrosis and false lobular formation.Compared with the model group,the levels of serum ALT,AST,ALP and TBIL were significantly decreased in the treatment group,and the levels of HA,LN,PCⅢand TIMP-1 in liver tissue were significantly decreased,and the high and low dose groups could significantly decrease the liver coefficient,While low-dose group can reduce the spleen coefficient;（P<0.05or P<0.01）,and the levels of TGF-β₁ in the low dose group were significantly lower than those in the control group（P<0.05 or P<0.01）The degree of fibrosis was improved in different degrees.3.Effects of Gansu Granule on bile duct obstructive hepatic fibrosis rats.Compared with the control group,the serum levels of ALT,AST,ALP,TBIL and DBIL were significantly increased in the model group,the liver coefficients were significantly increased,ALB content and white blood ratio were significantly decreased（P<0.05 or P<0.01）,HE showed mild to moderate fibrosis in liver tissue;Compared with the model group,the levels of serum ALT,AST,TBIL,ALP,DBIL and globulin were decreased and the liver coefficient was decreased.The colchicine group could decrease the spleen coefficient（P<0.05,P<0.01）ALB content and white blood ratio increased,colchicine group and liver and liver granules group of liver fibrosis and model group compared with the lesion has been reduced.Conclusions:Gansu granule is effective on hepatic fibrosis rats induced by methods of CCl₄,immunoreatction and bile duct obstruction respectively,which can significantly improve liver function and morphorlogically reduce hapatic fibrosis.These effects refers to that Gansu granule can reduce hepatic lipid peroxidation damage,inhibit th collagen deposition and improve collagen degradation in liver.Thus,synthesis of extracellular matrix and expressions ofα-SMA,TGF-β₁ were suppressed.