肝细胞生长因子阻抑TGF-β1诱导大鼠韧带成纤维细胞α-SMA过表达及其机制

Hepatocyte growth factor suppresses the overproduction of α-SMA induced by TGF-β1 in rat medial collateral ligament fibroblasts

目的:研究肝细胞生长因子(HGF)能否阻抑TGF-β1诱导的大鼠内侧副韧带(MCL)成纤维细胞α-SMA过表达及其可能涉及的信号传导通路。方法:采用组织块培养法培养大鼠MCL成纤维细胞,培养液中加入TGF-β1(5ng/ml)及HGF(10—40ng/ml)。培养72h后,用RT-PCR检测各组α-SMAmRNA及Smad3mRNA的变化;细胞免疫组化检测α-SMA蛋白的表达。结果:TGF-β1能显著诱导α-SMA及Smad3的表达(P<0.01),而HGF则可以有效地阻抑其表达,其效应呈剂量依赖性(P<0.05)。结论:HGF可以通过下调Smad3的表达来阻抑TGF-β1诱导的α-SMA过表达。这为利用HGF预防和治疗MCL损伤后瘢痕及纤维化在细胞和分子水平提供了依据。

Objective：To examine the effectiveness of HGF in blocking TGF-β1-induced α-SMA production in rat medial collateral ligament （MCL） fibroblasts. Method：Fibroblasts were obtained from rat MCL. Cell culture was supplemented with 5ng/ml of TGF-β1 along with increasing doses of HGF （10-40 ng/ml）. After 72 hours incubation, the productions of α-SMA and Smad3 mRNA were assayed by RT-PCR. Expression of α-SMA protein was assessed by immunostaining. Result：Treatment with TGF-β1 significantly stimulated α-SMA and Smad3 mRNA production in MCL fibroblasts （P〈0.01）. Remarkably, the addition of HGF reduced productions of all components induced by TGF-I31 in a dose-dependent manner （P〈0.05）. Conclusion：HGF antagonizes TGF-β1 induced α-SMA production in MCL fibroblasts by down regulating Smad3. The findings provide a cellular and molecular basis for HGF＇s acting as a therapeutic agent for MCL scar and fibrosis formation.