人血栓调节蛋白基因转染内皮祖细胞预防球囊扩张术后动脉再狭窄  

Transplantation of human thrombomodulin gene-modified endothelial progenitor cells to prevent vascular restenosis after balloon injury

作　　者：侯居攀[1] 吴志平[1] 金晖[1] 秦永林[1] 柏志斌[1] 柳东芳[1] 丁洁[1] 程科萍[1] 邓钢[1] 

机构地区：[1]东南大学附属中大医院介入与血管外科江苏省分子影像与功能影像重点实验室,南京210009

出　　处：《中华实验外科杂志》2013年第5期908-910,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金资助项目（81171433/H1816）;江苏省自然科学基金资助项目（BK2010395）

摘　　要：目的探讨血栓调节蛋白基因（hTM）转染的内皮祖细胞（EPCs）移植至兔颈总动脉球囊损伤局部预防动脉冉狭窄的可行性。方法兔外周血分离培养EPCs；脂质体介导基因转染，片以Fe2O3磁性纳米颗粒标记EPCs；高脂饮食联合球囊扩张制作兔双侧颈总动脉损伤模型共16只，随机分成2组，双腔Fogarty球囊阻断法分别将转染EPCs、未转染EPCs及等量生理盐水移植至兔颈总动脉损伤局部；移植后1、3、7、14d分别行磁共振（MR）扫描，观察损伤局部信号强度改变；4个月后行血管组织切片苏木素-伊红（HE）染色及弹力染色，观察再狭窄情况。结果脂质体可以成功地将外源hTM基因转染至EPCs并可获得10％～15％的转染效率；Fogarty球囊阻断法可以使部分EPCs黏附于血管损伤局部，引起局部MR信号改变；基凶修饰组、基因未修饰组、单纯损伤组内膜中膜比（N／M）分别为0．30±0．02、0．62±0．05、1．58±0．10，3组间差异有统计学意义（P〈0．01）。结论血栓调节蛋白基因转染内皮祖细胞可以有效预防球囊扩张术后动脉再狭窄。Objective To explore the feasibility of transplantation of human thrombomodulin （hTM） gene-modified endothelial progenitor ceils （EPCs） to rabbit carotid artery balloon injury model lo- cally to prevent vascular restenosis. Methods Rabbit peripheral blood mononuclear cells were isolated and EPCs were modified with hTM gene using X-tremeGENE HP DNA Transfection Reagent. Genetically modified EPCs were incubated with Fe2O3-arginine. Sixteen adult New Zealand white rabbits were divided into two groups, hTM gene-modified EPCs, non-gene modified EPCs and equal volume of normal saline were transplanted to the balloon injury local via Fogarty balloon catheter, respectively. Serial nuelear magnetie resonance （MRI） was performed on 1st, 3rd, 7th and 14th day after transplantation to assess the loss of MRI signal intensity at the injured sites. Four months later, the carotid arteries were isolated and cut for hematoxylin and eosin （HE） staining and elastic tissue staining to evaluate restenosis. Results The hTM gene could be successfully transfected into EPCs using Transfection Reagent and the efficiency rate was about 10%-15%. EPCs could be partly adhered to the injured site locally via Fogarty balloon catheter and result in the loss of magnetic resonance （MR） signal intensity at the injured sites. The neointimal/media ratio （N/M） in gene modified group, non-gene modified group and injured group was 0. 30 ± 0. 02, 0. 62 ± 0. 05 and 1.58 ± 0. 10, respectively （P 〈 0. 01）. Conclusion hTM gene-modified EPCs could effectively prevent vascular restenosis after balloon injury.

关 键 词：血栓调节蛋白 内皮祖细胞 球囊损伤 再狭窄 

分 类 号：R541.4[医药卫生—心血管疾病]