季德胜蛇药抗四氯化碳致小鼠肝纤维化的作用及机制研究

目的: 观察季德胜蛇药对四氯化碳(CCl₄)致小鼠肝纤维化的拮抗作用,探讨其抗纤维化的作用机制。方法: ICR小鼠48只随机分为正常对照组、模型组、季德胜蛇药(0.25,0.37 汪g·kg^-1)2组。除正常组外各组小鼠均以40%CCl₄ sc共5周。自造模日起,各治疗组小鼠每日分别给予季德胜蛇药ig,正常对照组及模型组小鼠每天灌胃等量容积蒸馏水。5周后,放射免疫法检测小鼠血清透明质酸(HA)和层黏蛋白(LN)水平;Masson三色染色法观察肝组织病变。制备季德胜含药兔血清,不同体积分数的含药血清培养肝星状细胞(HSC),CCK-8法检测HSC的增殖,流式细胞仪检测HSC凋亡。结果: 季德胜蛇药0.25,0.37 g·kg^-1组小鼠肝组织纤维化程度及计分均较模型组明显减轻(P<0.05);季德胜蛇药0.25 g·kg^-1组血清HA,LN水平均明显低于模型组(P<0.05,P<0.01),且0.37 g·kg^-1组均低于0.25 g·kg^-1组(P<0.01);与对照组比较,以10%和12.5%季德胜蛇药含药血清培养的HSC细胞的增殖率明显降低(P<0.05,P<0.01),而细胞凋亡率显著增加(P<0.01)。结论: 季德胜蛇药能抑制HSC的增殖和促进HSC凋亡,具有一定的抗肝纤维化作用。

Effect and its Mechanism of Jidesheng Sheyao on Hepatic Fibrosis Induced by Carbon Tetrachloride in Mice

Objective: To observe the protective effect of Jidesheng Sheyao, a Chinese patent medicine, on liver fibrosis of mouse caused by carbon tetrachloride(CCl₄) and to explore the action mechanism of the drug. Method: Forty-eight ICR mice were randomly divided into four groups:the normal control group, the model group, treatment group 1 and treatment group 2.All mice were subcutaneously injected with 40%CCl₄ for 5 weeks in order to prepare liver fibrosis model. Meanwhile, the mice of two treatment groups were given with Jidesheng Sheyao in different doses and the normal the mice of control group were given with distilled water in the same volume, by intragastric administration. The concentrations of hyaluronic acid (HA) and laminin (LN) in serum of mouse were measured with the method of radioimmunoassay. Masson trichrome staining was adopted to prepare hepatic tissue section. The pathological variations in hepatic tissue were observed under light microscope and the score of hepatic fibrosis was evaluated. The rabbits were given with Jidesheng Sheyao by intragastric administration to prepare medicated serum. The hepatic stellate cell strain (HSC-T6) was taken as the model for study in vitro. HSC cells were cultured with medicated serum in different concentrations. The proliferation of HSC was assessed with CCK-8 assay. Labeled with annexin V/PI, the apoptosis of HSC was analyzed with flow cytometry. Result: Compare with mice of model group, the severities of fibrosis in mice in treatment groups were obviously alleviative,the levels of fibrosis score and the serum content of HA, LN of mice in treatment groups were all lower. After cultured with the medicated serum respectively in the concentration of 10% and 12%, the proliferations of HSC were obviously inhibited and the apoptosis rates of the cells were significantly increased, compared with the cells of control group cultured with normal rabbit serum. Conclusion: Jidesheng Sheyao can inhibit the proliferation of HSC and promote the apoptosis of HSC, so the drug has some efficacy of anti-hepatic fibrosis.