肝康合剂实验性治疗大鼠肝纤维化的分子机制研究

Molecular mechanism of curing liver fibrosis with Gangkang mixture in experimentinduced liver fibrosis rat

目的:探讨肝康合剂对实验性免疫诱导型肝纤维化大鼠的治疗作用及机制。方法:建立雌性Wistar大鼠免疫损伤性肝纤维化大鼠模型,将肝康合剂作为抗肝纤维化药物,通过光镜观察肝脏HE、VanGieson(VG)胶原染色,原位杂交技术及免疫组化染色等方法,观察大鼠使用肝康合剂后肝脏组织中TIMP-1、TIMP-2的表达。结果:免疫诱导肝纤维化大鼠在造模结束后肝组织学改变呈进行性加重,以造模结束后3月为著,TIMP-1、TIMP-2mRNA及蛋白呈强阳性表达,电镜见模型组有较多激活的HSC及大量胶原纤维沉积在肝窦周间隙及肝细胞间,经肝康合剂治疗后的大鼠与模型组大鼠相比肝组织结构明显好转,纤维组织明显减少,未见到激活的HSC,TIMP-1、TIMP-2mRNA及蛋白表达与模型组相比均明显降低(P<0.05),并且其效果为预防组及治疗组效果均明显优于秋水仙碱组。结论:肝康合剂可以逆转肝纤维化,抗肝纤维化机制可为:1抑制肝星状细胞活化,减少ECM的生成及TIMP分泌;2直接抑制TIMP-1、TIMP-2mRNA和蛋白的表达;二者共同作用使得TIMP分泌减少,从而降低对MMPs的抑制,有利于MMPs对ECM的降解。

Objective ：To study the therapeutic effect and mechanism of Chinese traditionary medicine Gangkangheji using immune induced liver fibrosis rat model. Methods ： Rat liver fibrosis was induced by the administration of 200g/L human serum albumin （HSA） by iv eighty rats were divided into 5 groups. In group B, chinese medicine gankang mixture was taken orally at the same time with the administration of HSA. In group C, Chinese medicine gankang mixture was taken orally immediately after the administration of HSA. In group D, chinese medicine gankang mixture was taken orally at 3 months after the administration of HSA. In group E, colchine tablets was taken orally at the same time with the administration of HAS. In group F, HSA was administrated without treatment . Group A is as untreated control. The changes of HE. VG and expression of mRNA were observed with optical microscope. Moreover,the ultrastructure was observed with electron microscope. Results：In the immune induced liver fibrosis model rats, the fibrosis was most serious at 3 months after the administration of HAS. In the gankang mixtur treatment rats, group B is better than group C. The gankang mixture treatment was superior to colchine tablets treatment. Conclusion： Animal study has confirmed that gankang mixture decoction /capsule has the ability to prevent and cure liver fibrosis . The possible mechanism may be. ①by inhibiting the activation of HSC and the inhibiting the formation of TIMP and promoting the degradation of ECM; ②by inhibiting the mRNA expression of TIMP-1, TIMP-2 and protein expression to reduce the inhibition to MMPs and to enhance the degradation of ECM.