The Effect Of Ganbao Capsule On The Prevention And Treatment Of Liver Fibrosis Induced By DMN In Rats

Purpose:The aim of the study is to evaluate the effect of Ganbao capsule on prevention and treatment of liver fibrosis induced by Dimethyl nitrosamine (DMN) in rats and explore its mechanisms of action. Meanwhile, the internal mechanism of synergy between the single herbs in the prescription was investigated, which will provide experimental data for its clinical applications.Methods:Rats were induced liver fibrosis by intraperitoneally injection of DMN10μl·kg-1into rats once a day during continuous3days in a week, for4weeks. Meanwhile, the rats were Ganbao capsule1.0,2.0,4.0g·kg-1, or equivalent solvent. And the synergistic effect between Scutellaria and schisandra was investigated using factorial design.After24h of the last administration, the rat serum was taken to measure related biochemical indicators i.e. T-BiL, AST, ALT, ALP, y-GT, TP and ALB,a2MG, HPT and Apol. Meanwhile, the rat liver was weighed, and the content of Hyp, VEGF, IL-1α, IL-1β and IL-18were measured using protein suspended chip technology. The livers were observed using HE and Masson staining to check the change of liver histomorphology and collagen thickening. Immunohistochemical method was used to evaluate the expression of TGF-β1、α-SMA in liver tissue.Results:Comparison to the control, in model rats, the body weight decreased after1week of treatment, the TP and ALB values in serum were obviously decreased at the end of experiment. The levels of T-BiL, ALP, y-GT, AST, ALT in serum and HyP in liver tissue were pronouncedly increased. The content of IL-la and IL-1(3in liver tissue markedly decreased, whereas Leptin, VEGF and IL-18prominently increased. The fibrosis in rat liver was clearly observed after HE staining, and the ratio of collagenous tissue content was significantly increased.The body weight of DMN treated rat significantly increased in the administration of Ganbao capsules1.0,2.0g·kg-1for3weeks, and Ganbao capsules at various dosages could improve the DMN-induced rat weight after4weeks of administration. Ganbao capsule at different dosages significantly antagonist the decrease of the hepatic index induced by DMN in rats.Ganbao capsules1.0g·kg-1could increase level of ALB, decreased the levels of T-BiL, y-GT, AST and ALT in the serum in DMN treated rats.Ganbao capsule1.0g·kg-1significantly lowered the levals of T-BiL, AST, ALT in serum and HyP in liver tissue in DMN treated rats. Ganbao capsule4.0g·kg-1apparently decrease the levels of T-BiL, AST, ALT and y-GT in serum as well as HyP content in liver tissue in DMN treated rats. After HE staining, the improvement of fibrosis was observed after the administration of Ganbao capsule1.0,2.0and4.0g·kg-1compared with the model group. The collagen volume fraction (CVF) was obtained through Masson staining, and there were significant difference between Ganbao capsule at various dosages with the model group in terms of CVF values, suggesting they were capable of restricting the appearance of collagenous fiber in DMN-induced rat liver. Ganbao capsule2.0g·kg-1could promote the expression of IL-1β in DMN treated rat liver tissue, Ganbao capsule4.0g·kg-1significantly increased the level of IL-1β and IL-1α, whereas lowered the expression of IL-18in DMN treated rat liver. Ganbao capsule2.0,4.0g·kg-1significantly decreased the positive area value of TGF-β1expression in DMN-induced rats, which exhibited significantly different in comparison to model group.The results of synergistic effect of Schisandra and Scutellaria on prevention and treatment of liver fibrosis induced by DMN showed that Schisandra2.0g·kg-1apparently improved the rat weight after3weeks of administration, Ganbao capsule4.0g·kg-1and Schisandra2.0g·kg-1significantly increased the body weight of DMN treated rats. In addition, Ganbao capsule4.0g·kg-1and Schisandra2.0g·kg-1could obviously suppress the decrease of liver index induced by DMN. Ganbao capsule4.0g·kg-1significantly lowered the level of T-BiL, AST, ALT and γ-GT in serum and HyP content in liver tissue in DMN treated rat, Schisandra2.0g·kg-1significantly increased the ALB content and decreased the activity of T-BiL,γ-GT, AST and ALT in serum in DMN treated rat.After HE staining, the improvement of liver fibrosis was observed after the administration of Schisandra2.0g·kg-1or Ganbao capsule4.0g·kg-1in comparison to model group. The results of Masson staining showed that Ganbao capsule4.0g·kg-1, Schisandra2.0g·kg-1and2.0g·kg-1Scutellaria could significantly restrict appearance of the liver collagenous fiber induced by DMN. Compared with model groups, Schisandra2.0g·kg-1significantly increased IL-1β level and lowered the expression of Leptin in the lever in DMN treated rats. Schisandra2.0g·kg-1significantly lowered the positive area value of TGF-β1expression. Scutellaria2.0g·kg-1decreased the positive area value and integrated optical density of TGF-β1expression in the liver in DMN treated rats.Schisandra and Scutellaria exhibited synergistic effect on increasing the level of ALB in serum and decreasing the content of HyP in liver tissue, as well as show synergistic effect on decreasing the expression of IL-18and TGF-β1in liver tissue.Conclusion:The results of the experiments exhibited Ganbao capsule had prevention and treatment effect on the liver fibrosis induced by DMN. The mechanism of action of Ganbao capsule involved in suppression of expression of IL-18, TGF-β1and a-SMA as well as promotion of IL-1β and IL-1α expression, which would restrict activation and proliferation of HSC to decrease the generation of ECM.