中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
5期
925-927,封3
,共4页
裴长安%秦士勇%陈士辉%张曙光
裴長安%秦士勇%陳士輝%張曙光
배장안%진사용%진사휘%장서광
Nelin%血管平滑肌细胞%表型转化%细胞骨架%收缩
Nelin%血管平滑肌細胞%錶型轉化%細胞骨架%收縮
Nelin%혈관평활기세포%표형전화%세포골가%수축
Nelin%Vascular smooth muscle cells%Phenotypic modulation%Cytoskeleton
目的 观察Nelin对血管平滑肌细胞(VSMC)表型转化的影响.方法 血清饥饿诱导VSMC由合成型向收缩型转变,应用逆转录-聚合酶链反应(RT-PCR)技术检测不同表型VSMC的Nelin mRNA表达水平；转染Nelin-小干扰RNA(siRNA)沉默Nelin的表达,应用考马斯亮蓝染色法、乙酰胆碱(Ach)刺激法观察VSMC细胞骨架和收缩功能的变化.结果 Nelin mRNA在血清刺激组与血清饥饿24、48、72 h组表达量分别为:0.1940±0.0971、0.7114±0.1265、0.7270±0.1026、0.7841±0.1240,差异有统计学意义(P＜0.05).在血清饥饿培养过程中,VSMC细胞骨架由稀疏、模糊网状重构为均匀、致密束状,并重获收缩功能.抑制Nelin表达后,血清饥饿诱导的细胞骨架重构受阻,Ach诱发的细胞收缩消失.结论 Nelin参与VSMC骨架重塑并调节其收缩功能,对VSMC表型转化的调节、收缩表型的维持具有重要作用.
目的 觀察Nelin對血管平滑肌細胞(VSMC)錶型轉化的影響.方法 血清饑餓誘導VSMC由閤成型嚮收縮型轉變,應用逆轉錄-聚閤酶鏈反應(RT-PCR)技術檢測不同錶型VSMC的Nelin mRNA錶達水平；轉染Nelin-小榦擾RNA(siRNA)沉默Nelin的錶達,應用攷馬斯亮藍染色法、乙酰膽堿(Ach)刺激法觀察VSMC細胞骨架和收縮功能的變化.結果 Nelin mRNA在血清刺激組與血清饑餓24、48、72 h組錶達量分彆為:0.1940±0.0971、0.7114±0.1265、0.7270±0.1026、0.7841±0.1240,差異有統計學意義(P＜0.05).在血清饑餓培養過程中,VSMC細胞骨架由稀疏、模糊網狀重構為均勻、緻密束狀,併重穫收縮功能.抑製Nelin錶達後,血清饑餓誘導的細胞骨架重構受阻,Ach誘髮的細胞收縮消失.結論 Nelin參與VSMC骨架重塑併調節其收縮功能,對VSMC錶型轉化的調節、收縮錶型的維持具有重要作用.
목적 관찰Nelin대혈관평활기세포(VSMC)표형전화적영향.방법 혈청기아유도VSMC유합성형향수축형전변,응용역전록-취합매련반응(RT-PCR)기술검측불동표형VSMC적Nelin mRNA표체수평；전염Nelin-소간우RNA(siRNA)침묵Nelin적표체,응용고마사량람염색법、을선담감(Ach)자격법관찰VSMC세포골가화수축공능적변화.결과 Nelin mRNA재혈청자격조여혈청기아24、48、72 h조표체량분별위:0.1940±0.0971、0.7114±0.1265、0.7270±0.1026、0.7841±0.1240,차이유통계학의의(P＜0.05).재혈청기아배양과정중,VSMC세포골가유희소、모호망상중구위균균、치밀속상,병중획수축공능.억제Nelin표체후,혈청기아유도적세포골가중구수조,Ach유발적세포수축소실.결론 Nelin삼여VSMC골가중소병조절기수축공능,대VSMC표형전화적조절、수축표형적유지구유중요작용.
Objective To investigate the effect of Nelin on phenotypic transition of vascular smooth muscle cells (VSMC).Methods During reversion of synthetic VSMC to contractile phenotype induced by serum starvation,the expression of Nelin mRNA in different phenotypes was detected by reverse transcriptase-polymerase chain reaction (RT-PCR).VSMC was transfected by Nelin-small interfering RNA (siRNA) to knock-down the expression of Nelin,and then observe the effects on cvtoskeleton and contractibility by respectively coomassie brilliant blue dyeing and acetylcholine (ACh)-induction.Results The expression level of Nelin mRNA in VSMC Which were stimulated hy serum and serum starvation for 24,48,72 h was 0.1940 ± 0.0971,0.7114 ± 0.1265,0.7270 ± 0.1026,0.7841 ± 0.1240 respeetively(P ＜0.05).The cytoskeleton was remodulated from discrete and fuzzy network to well arranged and dense bundles after serum starvation.At the same time,ACh-induced contractility was observed.After Nelin expression was inhibited,reorganization of cytoskeleton by serum starvation was bloeked,and Ach-induced contraction was absent.Conclusion Nelin was involved in VSMC cytoskeleton reorganization and played an important role for VSMC remaining contractile phenotype and phenotypie transition.
