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目的:观察桂枝茯苓丸对大鼠肝纤维化的拮抗作用,并探讨可能的作用机制。方法:按照随机数字表将60只Wistar大鼠分为正常组8只(A组)和造模组52只,正常组sc予生理盐水3 mL.kg-1,造模组采用等量40%四氯化碳(carbontetrachloride,CCl4)复制肝纤维化模型,共6周。再随机将造模组大鼠分为模型组(B组)、桂枝茯苓丸低、中、高剂量组(C,D,E组)每组7只大鼠。A组与B组大鼠均予生理盐水9.0 mL.kg-1ig,C,D,E组分别给予桂枝茯苓丸0.45,0.9,1.8 g.kg-1.d-1 ig治疗,共4周,并在治疗期间继续使用CCl4维持。采用免疫组化(immunohistochemistry,IH)法及实时定量PCR(real-timequantitative PCR,Real-time PCR)法检测各组大鼠肝组织α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、转化生长因子β1(transforming growing factorβ1,TGF-β1)、结缔组织生长因子(connective tissue grower factor,CTGF)、Ⅰ型胶原(CollagenⅠ,C-Ⅰ)及Ⅲ型胶原(CollagenⅢ,C-Ⅲ)的蛋白及基因表达量。结果:模型组大鼠肝组织α-SMA,TGF-β1,CTGF,C-Ⅰ,C-Ⅲ蛋白和mRNA的表达较正常组显著增多(P<0.01),治疗组均能下调大鼠肝组织α-SMA,TGF-β1,CTGF,C-Ⅰ以及C-Ⅲ蛋白和mRNA的表达,其中以D组,E组减少α-SMA,C-Ⅰ,C-Ⅲ蛋白的表达明显(P<0.05或P<0.01),以E组降低TGF-β1,CTGF蛋白表达最为显著(P<0.05),以D组,E组降低α-SMA,TGF-β1,CTGF mRNA的表达显著(P<0.01),以E组减少C-Ⅰ,C-ⅢmRNA的表达最明显(P<0.01)。结论:桂枝茯苓丸混悬液可有效减少肝组织α-SMA,TGF-β1,CTGF,C-Ⅰ,C-Ⅲ蛋白和mRNA表达量,有良好的抗肝纤维化作用。
Objective: To observe the antagonism of Guizhi Fuling capsule on hepatic fibrosis in rats and to explore its possible mechanism. Methods: Sixty Wistar rats were divided into normal group (n = 8) and model group (n = 52) according to a random number table. The normal group was given normal saline (3 mL.kg-1) Carbon tetrachloride (CCl4) replication of liver fibrosis model, a total of 6 weeks. Rats in model group were randomly divided into model group (B group), Guizhi Fuling pill low, medium and high dose group (C, D, E group) each group of 7 rats. Group A and group B rats were given normal saline 9.0 mL.kg-1ig, C, D, E were given Guizhi Fuling pill 0.45,0.9,1.8 g.kg-1.d-1ig treatment, a total of 4 weeks , And continued to use CCl4 maintenance during treatment. The expression of α-smooth muscle actin (α-SMA) in liver of each group was detected by immunohistochemistry (IH) and real-time quantitative PCR (Real- , Transforming growth factorβ1 (TGF-β1), connective tissue grower factor (CTGF), collagen Ⅰ (C-Ⅰ) and collagen Ⅲ (C-Ⅲ) Protein and gene expression. Results: The expression of α-SMA, TGF-β1, CTGF, C-Ⅰ, C-Ⅲ protein and mRNA in the model group were significantly higher than those in the normal group (P <0.01) The expression of α-SMA, C-Ⅰ and C-Ⅲ in group D and group E were significantly decreased (P <0.05) (P <0.01 or P <0.01). The expression of CTGF and TGF-β1 in E group were lower than those in E group (P <0.01) , And the expression of C-ⅠmRNA and C-ⅢmRNA were decreased obviously in E group (P <0.01). Conclusion: Guizhi Fuling pill suspension can effectively reduce the expression of α-SMA, TGF-β1, CTGF, C-Ⅰ, C-Ⅲ protein and mRNA in liver tissue and has a good anti-hepatic fibrosis effect.