中药复方对肝纤维化大鼠细胞因子的影响

Effect of HanDanGanLe on the cytokines in fibrotic rats

目的 研究结缔组织生长因子(CTGF)mRNA与转化生长因子 β1 (TGFβ1)mRNA在免疫性大鼠肝纤维化中的表达状况及中药复方(汉丹肝乐)对其的影响作用。 方法 用猪血清腹腔内注射复制免疫损伤性肝纤维化模型,造模同时给予汉丹肝乐口服作为肝纤维化防治组;免疫攻击12周后处死动物。取肝作HE染色显示肝纤维化形成状况,原位杂交方法检测肝内CTGF mRNA、TGFβ1 mRNA表达,并对指标作半定量评分。 结果 12周后模型组大鼠形成典型的肝纤维化,肝内CTGF mRNA与TGFβ1 mRNA表达均显著增强,同时二者阳性分布部位相近,表达程度呈正相关性(r=0.799,P<0.05);汉丹肝乐组,肝纤维化程度明显减轻,CTGF mRNA和TGFβ1 mRNA表达的阳性指数分别为12.5±2.3和25.4±3.2,模型组分别为28.8±1.4和37.3±5.4,t=5.208、5.655,P<0.01。Spearman等级相关分析显示,CTGF mRNA表达与组织病理上肝纤维化变化呈正相关性(r=0.822、0.808,P<0.05)。 结论 CTGF与TGFβ1基因的增高表达与肝纤维化形成有密切关系;汉丹肝乐能有效抑制CTGF与TGFβ1基因的表达。通过在转录环节上阻断肝纤维化相关的细胞内信号转导途径,可能是该药物作用的重要分子机制之一。

Objectives To study the expression of connective tissue growth factor (CTGF) mRNA and transforming growth factor β1 (TGF β1) mRNA in immunity-induced liver fibrosis rats and the effect of HanDanGanLe on them. Methods Male wistar rats were given intraperitoneal injection of porcine serum twice a week. At the beginning, the rats in HanDanGanLe-treatment group were feed with HanDanGanLe for precaution. The rats were killed after twelve weeks, then CTGF mRNA and TGF β1 mRNA were detected in liver samples with in situ hybridization, and the formation of liver fibrosis was observed with HE stain. The semi-quantitative results of the two genes expression were analysed along with the stages of hepatic fibrosis. Results Typical liver fibrosis developed in the model group rats, and the positive stain of CTGF mRNA and TGFβ1 mRNA increased, which was distributed in the areas where fibrosis occurred. There was obvious correlation between the expression strength of CTGF mRNA and TGF β1 mRNA (γ = 0.799, P < 0.05). In the rats receiving HanDanGanLe, CTGF mRNA expression index decreased markedly (12.5 ± 2.3 vs 28.8 ± 1.4, t = 5.208, P < 0.01), so did TGF β1 mRNA expression index (25.4 ± 3.2 vs 37.3 ± 5.4, t = 5.655, P < 0.01). There was also significant correlation between the scores of CTGF mRNA expression and the stages of hepatic fibrosis (rs =0.822, 0.808 in the model group and HanDanGanLe-treatment group, P < 0.05). Conclusions The expression of CTGF mRNA and TGF β1 mRNA is correlated closely with hepatic fibrosis degree. HanDanGanLe can effectively prevent the expression of CTGF and TGFβ1. One of the mechanisms of the intervention may be its blocking the intracellular signalling pathways involved in liver fibrogenesis.