肝宝胶囊对胆管结扎大鼠肝纤维化的机理研究

Anti-fibrotic effects and mechanism of Ganbao Capsule on bile duct ligation( BDL) rats

目的:探讨肝宝胶囊对胆管结扎大鼠肝纤维化的药效及其机制。方法:雌性SD大鼠,假手术组仅游离肝门部胆总管,其它各组以胆管结扎的方法制备大鼠胆汁淤积性肝纤维化模型,恢复性饲养5d后随机分为假手术组、模型组、双环醇0.1 g/kg、水飞蓟素胶囊0.1 g/kg、肝宝胶囊1.0、2.0、4.0原生药g/kg组。每天灌胃给药1次,连续4周。假手术组和模型组给予等体积0.5%CMC-Na溶液。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、AST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-1α等细胞因子水平。结果:胆管结扎大鼠在造模4周后,其血清ALT、AST、TBIL、DBIL、γ-GT、ALP含量明显升高,A/G、ALB明显降低,肝脏指数显著增加,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-18、MIP-1α、TGF-β1含量升高,IL-1α、IL-4、IL-10及TNF-α含量降低,与假手术组比较差异均有统计学意义。肝宝胶囊1.0g/kg可显著降低胆管结扎大鼠血清AST、ALT、TBIL、DBIL、γ-GT、ALP活性,升高其血清ALB水平及A/G比值,降低其肝脏组织Hyp、IL-18、TGF-β1含量及胶原堆积,升高IL-1α含量。肝宝胶囊2.0g/kg可显著降低胆管结扎大鼠血清AST、ALT、TBIL、DBIL、γ-GT、ALP活性,升高其血清ALB水平及A/G比值,可显著降低肝脏组织Hyp及TGF-β1含量,升高其IL-4及TNF-α含量。肝宝胶囊4.0g/kg可显著降低胆管结扎大鼠血清AST、ALT、TBIL、DBIL、γ-GT、ALP活性,升高其血清ALB水平及A/G比值,降低肝脏组织Hyp、MIP-1α、IL-18、IL-6及TGF-β1含量,升高其IL-1α、TNF-α、IL-13及PDGF-BB含量。结论:肝宝胶囊能有效地减轻胆总管结扎诱导的肝纤维化大鼠的肝损伤及纤维化程度,其作用机制可能与其能够降低大鼠肝脏组织内细胞因子IL-18及TGF-β1水平,减少HSCs活化有关;也可能与其能够减少肝脏趋化因子MIP-1α的合成与分泌,从而阻断MIP-1α所致的恶性循环,使肝纤维化逐步好转有关。

Objective：To investigate the therapeutic effect and mechanism of Ganbao Capsule （GBC） on the liver fibrosis induced by bile duct ligation （BDL） in rats. Methods： Female SD rats were BDL to induce liver fibrosis, meanwhile the sham operation group rats were just opened the abdominal cavity, bile duct separated, but not ligated 5 days later, the BDL rats were randomly divided into five groups, and were ig bicyclol 0. 1 g/kg, silymarin 0.1 g/kg, Ganhao 1.0 g/kg, Ganbao 2.0 g/kg, Ganbao 4.0 g/kg or the same volume of 0, 5% CMC separately, once a day for 4 weeks. On the 29th day,the rats were sacrificed, the activities of ALT and AST, the levels of ALB, TP, LN in serum were measured, and the livers and spleens were weighted and the indexes were calculated, the livers were taken to detect the content of Hyp, IL- 10,IL-13, IL-12pT0, IL-4, IL-6,IL-12,IL-18,IL-1α, IL-1β,MCP-1,MIP-1α, MIP-2, TNF-α, PDGF-BB, IFN-γ/and TGF-β or the expression of a-smooth muscle actin （α-SMA）. Results：After BDL 4 weeks, the activities of ALT, AST, TBIL, DBIL, γ-GT and ALP were increased significantly, the content of ALB and A/G ratio were significantly reduced, and the index of liver, accumulation of collagen and expression of α-SMA in liver were enhanced. The content of hydroxyproline （Hyp）, IL-18, MIP-1α and TGF-β1 were increased significantly meanwhile the content of IL-1α, IL-4, IL-10 and TNF-α was reduced. GBC 1.0 g/kg significantly decreased the activities of AST, ALT, TBIL, DBIL, γ-GT and ALP in serum and content of HyP, IL-18,TGF-β1 in liver, increased the levels of ALB , A/G ratio in serum and content of IL-1α in BDL rats. GBC 2.0 g/kg significantly reduced the activities of AST, ALT, TBIL, DBIL, γ-GT, ALP in serum and content of HyP and TGF-β1 in liver, increased the levels of ALB,A/G in serum and content of IL-4 and TNF-α in the liver in BDL rats. GBC 4.0g,/kg significantly reduced the activities of AST, ALT, TBIL, DBIL, γ-GT, ALP in serum and content of HyP, MIP-1α, IL-18, IL-6 and TGF-β1 in liver, increased the levels of ALB, A/G in serum, the expression of IL-1α, TNF-α, IL-13 and PDGF-BB in liver in DBL rats. Conclusion： GBC have an protective effect on the liver injury and fibrosis induced by BDL. This might be attributed to GBC can down regularing the expression of TGF-β and IL-18 to inhibit the activation of HSC, or reduce the synthesis and secretion of ehemokines MIP-1α to cut off the vicious circle induced by increased MIP-1α. The detailed mechanism remains to be further study.