中药双甲五灵冲剂对免疫性肝纤维化大鼠肝组织中TIMPs蛋白及基因表达的影响

Effect of Chinese herb Shuangjiawuling on expression of TIMPs in rat with liver fibrosis

目的:研究在活血化淤基础上辅以软坚散节自制的双甲五灵冲剂对大鼠肝纤维化模型抗纤维化的治疗机制.方法:采用免疫诱导型肝纤维化大鼠模型,80只Wister大鼠,分为5组,A预防组造模同时开始喂食双甲五灵冲剂;B治疗组1在造模结束的同时开始喂食双甲五灵冲剂;C治疗组2在造模结束3mo后开始喂食双甲五灵冲剂;D秋水仙碱组在造模结束的同时开始喂食秋水仙碱;另设正常对照组10只.采用HE染色观察肝组织改变及Von-Gieson胶原纤维特殊染色以观察肝纤维化程度,并采用肝组织原位杂交及免疫组织化学染色观察大鼠肝脏组织中TIMP-1和TIMP-2mRNA及蛋白的表达强度.结果:免疫诱导型肝纤维化大鼠,在造模结束后肝组织学改变呈进行性加重,以造膜结束后3mo为著,其TIMP-1和TIMP-2mRNA及蛋白呈强阳性表达,而经双甲五灵冲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,TIMP-1和TIMP-2mRNA及蛋白表达明显降低(P<0.05),并且其效果为:预防组效果最好,治疗1组较治疗2组效果好,即造模同时开始喂食双甲五灵冲剂最好,造模结束同时用药好于造模结束3mo后用药,三组均明显优于秋水仙碱组.结论:中药双甲五灵冲剂治疗肝纤维化的机制可能通过抑制TIMP-1和TIMP-2mRNA及蛋白的表达而降低对MMPs的抑制,从而有利于MMPs对细胞外基质的降解.

AIM:To explore the effect of Chinese herb Shuangjiawuling in preventing hepatic fibrosis in rat with liver fibrosis. METHODS:Rat immunogenic liver fibrosis model was induced by administration human serum albumin (HSA 200 g/L) intravenously, 80 rats were randomly divided into 5 groups. Shuangjiawuling was taken orally simultaneously with (group A), immediately after (group B), or 3 months after ( group C) administration of HAS intravenously. Colchine tablets were taken orally with the administration of HAS (group D),normal rats were used as control ( Group E ). The pathological changes of liver tissue were observed by HE and Von-Gieson staining. The hepatic mRNA and protein of TIMP-1 and TIMP-2 were analyzed by in site hybridization and immunohistochemistry. RESULTS:Hepatic fibrosis developed gradually and became obviously on the third month after administration of HAS. And strong expressions of TIMP-1 and TIMP-2 were observed, however,its expression and hepatic fibrosis were reduced dramatically following administration of Shuangjiawuling and colchine tablets.The effect of Shuangjiawuling in preventing fi- brosis was superior to that of colchine tablets (P <0.05), and its effect in group A, B, and C decreased in order.CONCLUSION:Chinese herb Shuangjiawuling can inhibit the development of hepatic fibrosis and the expression of TIMPs, and promoted the effect of the degradation of the extracellular matrix.