TFPI基因转染对血管平滑肌细胞中细胞凋亡抑制蛋白的调控  

The Regulation on the Expression of Inhibitor of Apoptosis Proteins by TFPI Gene Transfer in Vascular Smooth Muscle Cells

作　　者：马丹丹[1] 李辉[1] 傅羽[1] 赵勇[2] 刘越[1] 尹新华[1] 

机构地区：[1]哈尔滨医科大学附属第一医院心内科,黑龙江省哈尔滨市150001 [2]哈尔滨医科大学附属第一医院普外科,黑龙江省哈尔滨市150001

出　　处：《中国动脉硬化杂志》2015年第8期769-773,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学青年基金项目(81200143;81200235);黑龙江省自然科学青年基金项目(QC2012C015);哈尔滨市科技局优秀学科带头人项目(2012RFXXS008);黑龙江省中医药管理局科技项目(ZHY12-W035)

摘　　要：目的研究组织因子途径抑制物(TFPI)基因转染对大鼠血管平滑肌细胞中细胞凋亡抑制蛋白(IAP)表达的影响,探讨TFPI诱导细胞凋亡的可能机制。方法将含有人TFPI基因的重组腺病毒或含β-半乳糖苷酶(LacZ)基因的重组腺病毒或DMEM在体外分别转染大鼠血管平滑肌细胞,用ELISA方法测定转染后细胞中TFPI蛋白的表达水平,用RT-PCR方法测定基因转染后不同时间点细胞中c-IAP1mRNA的表达,用Western-blot方法检测基因转染后不同时间点细胞中survivin的表达。结果基因转染后1天在血管平滑肌细胞中即可检测到TFPI蛋白表达,峰值出现在转染后第3天;基因转染后3天和7天,TFPI组c-IAP1 mRNA的表达与对照组相比明显减少(P<0.05);基因转染后3、5、7天,TFPI组survivin的表达与对照组相比明显减少(P<0.05),且具有明显的时间依赖性。结论 TFPI可能通过抑制IAP的表达来发挥诱导平滑肌细胞凋亡的作用,从而抑制冠状动脉介入治疗后再狭窄发生。Aim To investigate the effect of tissue factor pathway inhibitor（ TFPI） gene transfer on the expression of inhibitor of apoptosis proteins（ IAP） in vascular smooth muscle cell（ VSMC）. Methods Human TFPI or LacZ recombinant adenovirus or DMEM was used to infect rat aortic VSMC in vitro respectively. ELISA was used to examine the expression of exogenous TFPI gene. RT-PCR was used to detect the expression of c-IAP1 mRNA and western blot was used to detect the expression of survivin at different time points after gene transfer. Results TFPI protein expression was detected in VSMC at 1st day after gene transfer and the peak expression was at the 3rd day. The expression of cIAP1 mRNA in TFPI group was reduced compared with those in the Lac Z and DMEM groups（ P〈0. 05）. The expression of survivin in TFPI group was also decreased compared with those in Lac Z and DMEM groups（ P〈0. 05） in a time dependent manner. Conclusions TFPI might induce VSMC apoptosis through the inhibition on the expression of IAP,and therefore reduce the occurrence of restenosis after percutaneous coronary intervention.

关 键 词：基因转染 组织因子途径抑制物 血管平滑肌细胞 细胞凋亡抑制蛋白1 SURVIVIN 

分 类 号：R363[医药卫生—病理学]