肝宝胶囊抗免疫性肝纤维化及其机制的研究

Pharmacological study on the protective effects and its mechanism of Ganbao Capsule against immunohepatic fibrosis on a mouse model

目的:探讨肝宝胶囊对免疫性肝纤维化小鼠的治疗作用及机制。方法:选用全雌Balb/c小鼠,随机分为对照组、模型组、肝宝胶囊1.25、2.5、5 g生药/kg、双环醇0.1 g/kg,正常组小鼠尾静脉注射等体积无菌生理盐水溶液,其他各组尾静脉注射ConA 13 mg/kg造模,每周一次,连续7周;造模的同时分别灌胃药物或0.5%CMC-Na,每天一次,连续7周。末次给药后24小时,取肝脏、脾脏称湿重并计算其脏器指数,取血测定ALT等生化指标,取肝脏测定其羟脯氨酸、胶原堆积、α-SMA表达以及IL-10等细胞因子水平。结果:小鼠在注射ConA 13 mg/kg 7周后,其血清ALT、AST活性及TP、GLB、LN含量明显升高,A/G、ALB明显降低,肝脏指数和脾脏指数显著增加,肝脏组织羟脯氨酸(Hyp)、胶原堆积及α-SMA表达明显增加,肝脏IL-10、IL-12p70含量升高,而其IL-13、IL-4的含量降低,与对照组比较差异有统计学意义。肝宝胶囊1.25 g生药/kg可显著降低ConA小鼠血清ALT活性及GLB水平,升高其血清ALB水平及A/G比值,降低其肝脏羟脯氨酸含量及胶原堆积,显著降低小鼠肝脏组织的IL-10、IL-12p70、IL-13、TNFα、MCP-1以及MIP含量。肝宝胶囊2.5 g生药/kg可显著降低Con A小鼠血清ALT活性、GLB和LN水平,升高其血清ALB水平及A/G比值,降低其肝脏羟脯氨酸含量及胶原堆积,减少α-SMA表达,显著降低小鼠肝脏组织的IL-12p70、IL-13、TNFα、MCP-1以及MIP含量。肝宝胶囊5.0 g生药/kg可显著降低ConA小鼠血清ALT活性、GLB和LN水平,升高其血清ALB水平及A/G比值,降低其肝脏羟脯氨酸含量及胶原堆积,减少α-SMA表达,显著降低小鼠肝脏组织的IL-12p70、IL-13、TNFα、MCP-1、MIP以及TGFβ1含量。结论:上述研究结果表明肝宝胶囊防治ConA诱导小鼠的肝纤维化可能与其能够显著抑制和肝纤维化各阶段有关的细胞因子表达有关,而其抑制IL-10表达的作用,提示该药可能对病毒引起的慢性肝炎及肝纤维化有较好的疗效,其详细作用机理还有待进一步深入研究。

Objective： To investigate the therapeutic effect and mechanism of Ganbao Capsule against immunohepatic fibrosis. Methods： Female Balb/c mice were divided into 6 groups randomly：normal control group,ConA 13 mg/kg,bieyelol 0.1 g/kg, Ganbao 1.25 g/kg, Ganbao 2. 5 g/kg and Ganbao 5 g/kg . Mice were injection of 13 mg/kg or the same volume of normal saline once a week for 7 weeks. Meanwhile the mice ig different concentration of Ganbao Capsule（ GBC）, bieyelol or the same volume of 0.5% CMC-Na, once perday for 7 weeks. 24 h after the treatment, the livers and spleens were weighted and the indexes were calculated, the activities of ALT and AST, the levels of ALB, TP, LN in serum were measured, the content of Hyp, IL-10, IL-13, IL-12p70, IL-4, IL-5, MCP-1, MIP, TNF-α and TGF-β1 or the expression of a-smooth muscle aetin （ct-SMA） in the livers were detected in mice. Results ： After injection of ConA 13 mg/kg for 7 weeks, the activities of ALT and AST, the levels of TP, GLB and LN in serum was increased obviously, and the levels of ALB and A/G were obviously decreased in ConA treated mice. The coetfieients of liver and spleen, the content of Hyp, IL-10 and IL-12p70, the accumulation of collagen and the expression of α-SMA were obviously increased in liver in ConA treated mice . However, the content of IL-13, IL-4 in liver were decreased. GBC 1.25 g/kg could obviously decrease the activity of ALT,the level of GLB in serum,and the content of Hyp,IL-10,IL-12p70, IL-13 ,TNFα,MCP-1 ,MIP, and the accumulation of collagen in liver of ConA treated mice. This dosage also obviously increased the levels of ALB and A/G in serum in ConA treated mice. GBC 2.5 g/kg could significantly decrease the activity of ALT, the levels of GLB and LN in serum,increase the level of ALB in serum, decrease the concentration of Hyp, IL-12p70, IL-13,TNFct, MCP-1, MIP, the aeeumulation of collagen and the expression of α-SMA in liver in ConA treated mice. GBC 5.0 g/kg could significantly decrease the activity of ALT, the levels of GLB and LN in serum, increase the levels of ALB and A/G in serum, decrease the content of Hyp, IL-12p70, IL-β, TNFα, MCP-I, MIP and TGFβ1 ,the accumulation of collagen and the expression of α-SMA in liver in ConA treated mice. Conclusion： The protective effects of GBC on ConA-induced immunohepatic fibrosis may be related to inhibition of the expression of cytokines which have something to do with liver fibrosis. The action of decreasing the expression of IL-IO indicates that GBC may have an effect on virus chronic hepatitis and liver fibrosis. The detailed mechanism remains to be further in-depth study.