复方鳖甲软肝片对肝纤维化大鼠肝脏低氧诱导因子1α及其下游分子mRNA表达的影响

Effects of Fufang Biejia Ruangan Pian on mRNA expression of hypoxia-inducible factor-1α and its downstream molecules in livers of rats with hepatic fibrosis

[目的]观察复方鳖甲软肝片对肝纤维化大鼠肝组织低氧诱导因子1α(HIF-1α)及其下游分子血管内皮生长因子(VEGF)及结缔组织生长因子(CTGF)mRNA表达的影响,探讨复方鳖甲软肝片抗肝纤维化的作用机制。[方法]采用40%CCl4诱导大鼠肝纤维化模型,同时以1g/(kg.d)的复方鳖甲软肝片灌胃干预6周,常规苏木精-伊红和Masson染色观察大鼠肝组织炎症和纤维化程度,RT-PCR法检测肝组织HIF-1α、VEGF、CTGF基因mR-NA表达。[结果]模型组大鼠肝组织炎症和纤维化程度计分较正常组明显增高(P<0.01),肝组织HIF-1αmR-NA、VEGF mRNA及CTGF mRNA的表达较正常组明显增强(P<0.01,<0.05),肝组织纤维化程度与HIF-1αmRNA、VEGF mRNA及CTGF mRNA表达均呈明显正相关(均<0.01),肝组织HIF-1αmRNA与VEGF mR-NA、CTGF mRNA表达也呈明显的正相关(均P<0.01);应用复方鳖甲软肝片干预后,大鼠肝组织炎症和纤维化程度显著减轻(P<0.01,<0.05),肝组织HIF-1αmRNA、VEGF mRNA及CTGF mRNA表达明显下降(P<0.05)。[结论]复方鳖甲软肝片下调肝纤维化大鼠肝组织HIF-1α及其下游分子mRNA表达可能是其抗肝纤维化作用的机制之一。

[Objective]To evaluate the effectiveness of Fufang Biejia Ruangan Pian on mRNA expression of hypoxia-inducible factor-1 （HIF-1a）and its downstream molecules vascular endothe- lial growth factor（VEGF）and connective tissue growth factor（CTGF） in rats with hepatic fibro- sis, and to explore its possible mechanism of anti-hepatic fibrosis. I-Methods]Rats were subcutane- ously given injection of 40% carbon tetrachloride （3 ml·kg-1· d-1, twice every week for 6 weeks, double in the first dose）to induce the model of hepatic fibrosis, and were prevented by Fu- fang Biejia Ruangan Pian（1g· kg-1 · d-l）. The degrees of rat liver inflammation and fibrosis were observed by HE and Masson staining,while the mRNA expression of HIF-1a, VEGF and CTGF in liver were measured by reverse transcription polymerase chain reaction. [Results]The degrees of liver inflammation and hepatic fibrosis in the model group were higher than that in the normal group（P^0.01）. The hepatic HIF-la, VEGF and CTGF gene expression in the model group was higher than those in the normal group（P〈0.01, P%0.05）. The degree of hepatic fibrosis was positively correlated with the mRNA expression of HIF-1α , VEGF and CTGF（All P 〈0.01）. HIF-1α mRNA expression was also positively correlated with VEGF and CTGF mRNA expression（All P〈0.01）. After treatment with Fufang Biejia Ruangan Pian, the degrees of liver inflammation and hepatic fibrosis were remarkably depressed（P%0.01,P〈0.05）, and the mRNA levels of HIF-1α, VEGF and CTGF were obviously decreased（P〈0.05）. [Conclusion]Fufang Biejia Ruangan Plan can obviously depress the expression of hepatic HIF-1α and its downstream molecules,and it may be one mechanism for Fufang Biejia Ruangan Pian to treat hepatic fibrosis.