RhoA／ROKα反义寡核苷酸抑制TGF-β1诱导的血管外膜成纤维细胞α-SM-actin表达  

Antisense Oligonucleotides against RhoA-ROKαSignaling Pathway Attenuatesα-SM-actin Expression Induced by TGF-β1 in Adventitiai Fibroblasts

作　　者：陈闻东[1] 朱鼎良[1] 姬开达[1] 高平进[1] 

机构地区：[1]上海市血管生物学重点实验室,上海交通大学医学院附属瑞金医院,上海市高血压研究所,医学基因组学国家重点实验室,上海市200025

出　　处：《中国分子心脏病学杂志》2006年第5期272-275,共4页Molecular Cardiology of China

基　　金：科技部973项目<国家重点基础研究发展规划>(G2000056904)

摘　　要：目的α-平滑肌-肌动蛋白(a-SM-actin)的表达是血管外膜成纤维细胞/肌成纤维细胞表型转化的分子标记。本研究观察阻断RhoA-ROKα信号转导通路对于转化生长因子β1(transfor- ming growth factorβ1,TGF-β1)诱导的血管外膜成纤维细胞α-SM-肌动蛋白(actin)表达的影响,以揭示RhoA-ROKα信号通路在血管外膜成纤维细胞表型转化为肌成纤维细胞过程中的作用。方法使用贴壁法体外培养大鼠胸主动脉外膜成纤维细胞；用Western blot技术测定RhoA和ROKα在血管外膜成纤维细胞表型转化为肌成纤维细胞过程中的表达。结果RhoA-ROKα在血管外膜成纤维细胞表达,TGF-β1可以诱导RhoA表达上调。用反义寡核苷酸技术抑制RhoA或ROKα的表达后能够抑制α-SM-actin的表达。结论RhoA-ROKα信号转导通路参与了TGF-β1诱导的血管外膜成纤维细胞表型转化为肌成纤维细胞的过程。Objective Adventitial fibroblast differentiation to myofibroblasts plays an important role in vascular remodeling.Myofibroblasts proliferate,migrate and may participate in neointima formation post- vascular injury.But the molecular events underling the myofibroblast differentiation remain unknown.The aim of this study was to investigate the role of RhoA-ROKαsignaling pathway in adventitial fibroblast differ- entiation to myofibroblasts.Methods Myofibroblast differentiation was carried out by transforming growth factor 131(TGF-β1,10ng/ml)treatment to cultured adventitial fibroblasts for 24 hours.We used Western blot to detect the RhoA andα-smooth muscle-actin(α-SM-actin)protein expression in cultured adventitial fibroblasts/myofibroblasts.Results TGF-β1 up-regulated total RhoA protein expression,which peaked at 6 hours post-TGF-β1 stimulation.Interestingly,the time course of RhoA protein up-regulation was prior to the time course ofα-SM-actin protein up-regulation,the marker of myofibroblast differentiation,which peaked at 24 hours post-TGF-β1 stimulation.Antisense oligonucleotides against RhoA or ROKαbut not sense or mis- match oligonucleotides attenuated RhoA/ROKαas well asα-SM-actin expression.Conclusion RhoA- ROKαsignaling pathway participated inα-SM-actin expression induced by TGF-β1.RhoA-ROKαsignaling pathway may be required for adventitial fibroblast differentiation to myofibroblasts.

关 键 词：RHOA Rho-associated KINASE alpha 肌成纤维细胞 转化生长因子β1 反义寡核苷酸 

分 类 号：R543[医药卫生—心血管疾病]