连接蛋白Cx43参与冠状动脉平滑肌细胞的异质性促进冠脉再狭窄  

The role of connexin43 in coronary artery smooth muscle cells heterogeneity and coronary restenosis

作　　者：张旭敏[1] 马晓烨[1] 王小东[1] 姚义安[1] 

机构地区：[1]同济大学附属东方医院心内科,上海200120

出　　处：《中国实验诊断学》2014年第5期700-704,共5页Chinese Journal of Laboratory Diagnosis

基　　金：上海市浦东新区科委课题(PKJ2011-Y21)

摘　　要：目的研究猪冠状动脉平滑肌细胞异质性与连接蛋白Cx43关系,进一步阐明连接蛋白Cx43在冠状动脉再狭窄发生中的作用。方法分别培养正常及支架术后猪原代冠状动脉平滑肌细胞,正常组应用血小板源生长因子(PDGF-BB)进行诱导,两组细胞分别进行电子显微镜观察细胞形态及结构变化,采用Western Blotting方法及实时荧光定量PCR测定Cx43、Cx40、α-SMA、S100A4蛋白和mRNA表达;然后诱导组应用Cx43阻断剂进行干预,再次检测上述指标变化。结果正常冠状动脉平滑肌细胞可见:纺锤型(spindle-shaped,S-SMC)和长菱形(rhomboid,R-SMC)两类,以S-SMC为主,其内Cx40、α-SMA有较高的蛋白和mRNA表达,而Cx43表达较少;正常组经PDGF-BB诱导后细胞由S-SMC向R-SMC转变,同时伴随Cx43表达上调,而Cx40表达明显下降;通过反义RNA降低Cx43表达,这种变化受抑制,并且S-SMC细胞保持原有形态,同时表达a-肌动蛋白,支架组平滑肌细胞以R-SMC为主,Cx43、S100A4有较高的蛋白和mRNA表达。结论冠状动脉平滑肌细胞表型变化与连接蛋白Cx43表达密切相关,连接蛋白Cx43可能参与冠脉再狭窄的过程。Objective To investigate the correlation of connexin43 （Cx43）and coronary artery smooth muscle cells （SMC）heterogeneity,and the role in coronary restenosis.Methods SMCs from normal porcine coronary artery media and stent-induced intimalthickening cultured in vitro were induced by PDGF-BB and then intervened by Cx43 inhibitor. The cell morphology was observed by electron microscopy.The expression of Cx43、Cx40、α-SMA、S100A4 in SMCs of coronary artery were examined by Western Blotting and real time RT-PCR.Results The normal SMCs isolated from the porine coronary artery exhibit distinct phenotypes in vitro：spindle-shaped （S-SMC）and rhomboid （R-SMC）.S-SMCs were predominant in the normal media in which Cx40 andα-SMA was highly expressed ,whereas Cx43 was bare-ly detectable.PDGF-BB increased the level of Cx43,and promoted the transition of S-SMCs to R-SMCs.However,in-hibiting the expression of Cx43 decreased the level of Cx43,promoted the transition of R-SMCs to S-SMCs and ex-pressedα-SMA.R-SMCs were predominant in the stent-induced intimal thickening and that this modulation was accom-panied by the expression of Cx43 and S100A4.Conclusion The phenotype of coronary artery SMCs is closely related to the changes of the expression of Cx43 which may participate in the restenotic process.

关 键 词：支架内再狭窄 平滑肌细胞 连接蛋白 细胞表型 

分 类 号：R541.4[医药卫生—心血管疾病]