中国实验诊断学
中國實驗診斷學
중국실험진단학
CHINESE JOURNAL OF LABORATORY DIAGNOSIS
2014年
5期
700-704
,共5页
张旭敏%马晓烨%王小东%姚义安
張旭敏%馬曉燁%王小東%姚義安
장욱민%마효엽%왕소동%요의안
支架内再狭窄%平滑肌细胞%连接蛋白%细胞表型
支架內再狹窄%平滑肌細胞%連接蛋白%細胞錶型
지가내재협착%평활기세포%련접단백%세포표형
restenosis%connexin%cell phenotype%vascular smooth muscle cell
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。
目的：研究豬冠狀動脈平滑肌細胞異質性與連接蛋白 Cx43關繫，進一步闡明連接蛋白 Cx43在冠狀動脈再狹窄髮生中的作用。方法分彆培養正常及支架術後豬原代冠狀動脈平滑肌細胞，正常組應用血小闆源生長因子（PDGF-BB）進行誘導，兩組細胞分彆進行電子顯微鏡觀察細胞形態及結構變化，採用 Western Blotting 方法及實時熒光定量 PCR 測定 Cx43、Cx40、α-SMA、S100A4蛋白和 mRNA 錶達；然後誘導組應用 Cx43阻斷劑進行榦預，再次檢測上述指標變化。結果正常冠狀動脈平滑肌細胞可見：紡錘型（spindle-shaped，S-SMC）和長蔆形（rhomboid，R-SMC）兩類，以 S-SMC 為主，其內 Cx40、α-SMA 有較高的蛋白和 mRNA 錶達，而 Cx43錶達較少；正常組經 PDGF-BB 誘導後細胞由 S-SMC 嚮 R-SMC 轉變，同時伴隨 Cx43錶達上調，而 Cx40錶達明顯下降；通過反義 RNA 降低 Cx43錶達，這種變化受抑製，併且 S-SMC 細胞保持原有形態，同時錶達 a-肌動蛋白，支架組平滑肌細胞以 R-SMC 為主，Cx43、S100A4有較高的蛋白和 mRNA 錶達。結論冠狀動脈平滑肌細胞錶型變化與連接蛋白 Cx43錶達密切相關，連接蛋白 Cx43可能參與冠脈再狹窄的過程。
목적：연구저관상동맥평활기세포이질성여련접단백 Cx43관계，진일보천명련접단백 Cx43재관상동맥재협착발생중적작용。방법분별배양정상급지가술후저원대관상동맥평활기세포，정상조응용혈소판원생장인자（PDGF-BB）진행유도，량조세포분별진행전자현미경관찰세포형태급결구변화，채용 Western Blotting 방법급실시형광정량 PCR 측정 Cx43、Cx40、α-SMA、S100A4단백화 mRNA 표체；연후유도조응용 Cx43조단제진행간예，재차검측상술지표변화。결과정상관상동맥평활기세포가견：방추형（spindle-shaped，S-SMC）화장릉형（rhomboid，R-SMC）량류，이 S-SMC 위주，기내 Cx40、α-SMA 유교고적단백화 mRNA 표체，이 Cx43표체교소；정상조경 PDGF-BB 유도후세포유 S-SMC 향 R-SMC 전변，동시반수 Cx43표체상조，이 Cx40표체명현하강；통과반의 RNA 강저 Cx43표체，저충변화수억제，병차 S-SMC 세포보지원유형태，동시표체 a-기동단백，지가조평활기세포이 R-SMC 위주，Cx43、S100A4유교고적단백화 mRNA 표체。결론관상동맥평활기세포표형변화여련접단백 Cx43표체밀절상관，련접단백 Cx43가능삼여관맥재협착적과정。
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.
