| Background Ganshuang Granule(GS)is a kind of Chinese patent medicine for the treatment of liver fibrosis.It has been widely used in the treatment of liver fibrosis caused by various liver diseases,and has the effect of reducing liver inflammation and reducing the degree of liver fibrosis.However,there are few studies on how Ganshuang Granules play an anti-fibrosis role in animal experiments,and the mechanism of action is not clear yet.Studies have reported that the imbalance of intestinal flora runs through the beginning and end of the development of liver disease,and the abundance and structure of intestinal flora,as well as the metabolism of bacteria in liver fibrosis,are disorderly.The further aggravation of intestinal inflammation leads to the development of liver fibrosis towards cirrhosis.Therefore,on the basis of the entero-hepatic axis,this study explored how Ganshuang Granules could achieve the purpose of treating liver fibrosis by regulating intestinal flora.MethodsIn this study,firstly,the influence of Ganshuang Granules on the degree of fibrosis in liver fibrosis mouse model was explored.Secondly,the changes of inflammatory factors and peroxidase in serum were studied to clarify the mechanism of inflammation.In addition,16 S r RNA sequencing technology was used to detect the feces of mice in each group after modeling and drug treatment,so as to determine the abundance and structural changes of intestinal microflora in each group.Finally,the expression of tight junction protein in intestinal mucosa was studied by immunohistochemical method to observe the change of intestinal mucosa permeability.Results1 The results of HE staining,Masson staining and α-SMA staining showed that,compared with the model group,liver cell necrosis and inflammatory cell infiltration were significantly reduced after treatment with Gan Shuang Granules,and the degree of liver fibrosis was gradually reduced with the increase of therapeutic drug concentration.Serum test results of each group showed that liver index,aspartate aminotransferase(AST)level and alanine aminotransferase(ALT)level were significantly decreased in Ganshuang Granules treatment group.Ganshuang Granule has obvious therapeutic effect on liver fibrosis in model mice.2 ELISA and q PCR were used to detect the serum and liver tissue of each group.The results showed that compared with model group,the expression levels of interleukin(IL-1β,IL-6) and tumor necrosis factor(TNF-α)in each dose group of Ganshuang Granule decreased.The activities of glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD)in liver were increased,and the level of methane-dicarboxylic aldehyde(MDA)was decreased with the increase of dose in the treatment group.3 The results of 16 S r RNA sequencing analysis on the feces of mice in each group showed that the α diversity of intestinal microflora increased and the β diversity gradually recovered after Ganshuang Granules treatment compared with the model group;Compared with the model group,the F/B ratio of Ganshuang Granules medium-dose and high-dose groups was significantly decreased;At the subclass level,Lactobacillus,Akkermansia,Bacteroides and Oscillospira were all higher than those in the model group.Allobaculum and Acinetobacter were lower than those in model group.4 Immunohistochemical staining of colon in each group showed that Occludin and ZO-1 protein in each group of Ganshuang Granules were increased compared with that in the model group,and the expression levels of the two proteins increased gradually with the dosage of Ganshuang Granules increasing.Conclusions1 Ganshuang Granules can inhibit inflammation,reduce lipid peroxidation and reduce the severity of liver fibrosis in liver fibrosis model mice.2 Ganshuang Granules can regulate the diversity and abundance of intestinal microflora in liver fibrosis model mice.3 Ganshuang granules can reduce the permeability of intestinal mucosa and strengthen the barrier function of intestinal mucosa in liver fibrosis model mice. |
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