RhoA/SRF信号通路介导Nelin诱导人血管平滑肌细胞表型转化  

Gene Nelin induced phenotypic transition of human vascular smooth muscle cells in vitro mediated by RhoA and SRF

作　　者：裴长安[1] 秦士勇[1] 王明海[1] 张曙光[1] 

机构地区：[1]山东大学附属千佛山医院血管外科,济南250014

出　　处：《中华普通外科杂志》2014年第12期908-911,共4页Chinese Journal of General Surgery

基　　金：山东省自然科学基金资助项目（ZR2009CL018）

摘　　要：目的 探讨Nelin基因对人血管平滑肌细胞（vascular smooth muscle cell,VSMC）表型转化的调控机制.方法 应用过表达慢病毒载体[Nelin-VSMC]及干扰慢病毒载体[LV-Nelin-SiRNA-VSMC]制备稳定转染的VSMC细胞模型,通过荧光定量PCR以及蛋白质印迹分析（Western blotanalysis,WB）等技术手段,观察Nelin蛋白过表达及表达抑制对人VSMC表型转化的影响及其调控机制.结果 Nelin-VSMC组细胞呈收缩表型,细胞细长,成“谷-峰”样生长,Nelin mRNA、Nelin蛋白和平滑肌分化标志蛋白平滑肌α-肌动蛋白（SMoα-actin）表达显著增加,同时SMα-actin相关调控因子-血清反应因子（SRF）入核转位,RhoA总蛋白表达上调,经Rho激酶特异性抑制剂Y-27632作用后,SRF在细胞核中表达显著减少,同时SMα-actin表达下调;LV-Nelin-SiRNA-VSMC组细胞呈合成表型,细胞体积变大,细胞极性消失,生长状态无序,Nelin mRNA、Nelin蛋白和SMα-actin蛋白表达显著降低,同时伴有SRF出核转位及RhoA总蛋白表达下调.结论 在体外培养的人VSMC中,Nelin依次激活SMα-actin蛋白相关调控因子RhoA和SRF引起SMα-actin表达增加,促进VSMC向收缩表型转换.Objective To explore the mechanism of gene Nelin on the differentiation of vascular smooth muscle cells（VSMC）.Methods Human VSMCs were stably infected with over-express lentiviral vector [Nelin-VSMC] and interference lentiviral vector [LV-Nelin-SiRNA-VSMC] by Western blotting and RT-PCR.The effect of Nelin expression on phenotypic transition of VSMCs was observed and the mechanism involved was explored.Results The Nelin-VSMC cells take on the contractile phenotype of long slender cells possessing ＂hill-valley＂ characteristics.Overexpression of Nelin can enhance SMoα-actin expression with the increase of total RhoA and nuclear serum response factor（SRF） expression.Treated with a selective Rho kinase inhibitor Y-27632,the expression of SMα-actin and nuclear SRF protein induced by Nelin was effectively reduced.Meanwhile,the opposite results were observed in the Nelin-depression VSMC.Conclusions Nelin can enhance the expression of SMα-actin and carry a pivotal role in the process of VSMCs differentiation and phenotype modulation by activating Rho and SRF.

关 键 词：基因表达调控 肌细胞 平滑肌 Nelin基因 

分 类 号：R543[医药卫生—心血管疾病]