| To observe the effect of Yiguanjian(YGJ) combined with bone marrow mesenchymal stem cells(BMSCs) on Ras homolog gene family,member A(RhoA)/Rho-associated coiled-coil containing protein kinase 1(ROCK1) signaling pathway in the treatment of liver fibrosis.Methods:Seventy-two SD rats were randomly assigned into a normal group,a model group,a colchicine group,high-,medium,and low-dose YGJ(YGJ-H,YGJ-M,and YGJ-L,respectively) groups,a BMSCs group,and a YGJ+BMSCs group.The rat model of liver fibrosis was established by intraperitoneal injection of carbon tetrachloride(CCl4) for 6 weeks,and the rats were sacrificed after 4 weeks of treatment.Before the rats were sacrificed,we scored their behavior of yin deficiency.We then measured the content of glutamic-pyruvic transaminase(GPT) and glutamic-oxaloacetic transaminase(GOT) in the serum and stained the liver with hematoxylin-eosin(HE) and Masson to observe the pathological changes.RT-PCR and Western blotting were respectively employed to determine the mRNA and protein levels of RhoA,ROCK1,α-smooth muscle actin(α-SMA),and collagen type I(COL-1).Immunohistochemistry(IHC) was adopted to detect α-SMA and COL-1 positive areas in liver tissue sections.Results:Compared with the normal group,the modeling of liver fibrosis elevated the behavior scores of yin deficiency,caused liver inflammation and fibroplasia,up-regulated the mRNA and protein levels of RhoA,ROCK1,α-SMA,and COL-1(P<0.05),and increased the positive areas of α-SMA and COL-1(P<0.05).Compared with the model group,YGJ-H,YGJ-M,YGJ-L,BMSCs,and YGJ+BMSCs relieved the behaviors of yin deficiency,liver inflammation,and fibrosis(P<0.05),down-regulated the mRNA and protein levels of RhoA,ROCK1,α-SMA,and COL-1(P<0.05),and decreased the positive areas of α-SMA and COL-1(P<0.05).Among different treatment groups,YGJ+BMSCs group demonstrated the best performance(P<0.05).Conclusion:YGJ and BMSCs can inhibit the activation of hepatic stellate cells by inhibiting RhoA/ROCK1 signaling pathway to treat liver fibrosis,and their combination has better performance than YGJ or BMSCs alone. |
