基础医学与临床 ›› 2008, Vol. 28 ›› Issue (7): 756-759.

• 研究论文 • 上一篇    下一篇

TIMP-3基因瞬时表达对兔血管平滑肌细胞增殖、迁移及凋亡的影响

陈巾杰 浦晓东   

  1. 福建医科大学 福建医科大学
  • 收稿日期:2007-05-28 修回日期:2007-09-20 出版日期:2008-07-25 发布日期:2008-07-25
  • 通讯作者: 陈巾杰

Effections of TIMP-3 Gene Transient Expression on Proliferation,Migration and Apoptosis of Rabbit Vascular Smooth Muscle Cells

Jin-jie CHEN, Xiao-dong PU   

  • Received:2007-05-28 Revised:2007-09-20 Online:2008-07-25 Published:2008-07-25
  • Contact: Jin-jie CHEN,

摘要: 目的 通过阳离子脂质体介导TIMP-3基因质粒转染体外兔血管平滑肌细胞,观察基因瞬时表达对细胞增殖迁移及凋亡的影响,探讨TIMP-3基因治疗冠心病支架治疗再狭窄(ISR)的前景。方法 将细胞分为正常对照组、空质粒组、重组质粒载体组进行基因转染。1.转染后24小时、48小时、72小时对各组细胞进行计数、检测细胞MMP-2含量、TIMP-3mRNA水平。2.转染后48小时检测各组细胞凋亡率。3.建立细胞迁移模型,计算转染后48小时细胞迁移数。结果 正常对照组细胞生长、MMP-2含量、TIMP-3mRNA表达情况、细胞凋亡率、细胞迁移数与空质粒组相比均无明显差别;重组质粒组各项指标与前两者相比均有显著差别,P〈0.05。结论 TIMP-3对体外血管平滑肌细胞具有明显的抑制增殖、抑制迁移及促进细胞凋亡的作用;阳离子脂质体使用安全, 应用于TIMP-3基因治疗ISR前景广阔。

关键词: 特异性金属蛋白酶组织抑制剂-3, 血管平滑肌细胞, 阳离子脂质体, 基因转染, 再狭窄

Abstract: Objective Observe effections of TIMP-3 on proliferation, migration and apoptosis of rabbit vascular smooth muscle cells(VSMC)transfected by recombinant plasmid vectors mediated by cationic liposome and then discuss the TIMP-3 gene therapy on in-stent restenosis(ISR). Methods Divide VSMC into three groups as normal control group, empty plasmid group and recombinant group,and then do transfection separately.First, at time of 24hours, 48hours and 72hours , calculate cells ,quantify MMP-2 and analyse TIMP-3 mRNA. Second, measure cell apoptosis rate after 48hours. Third, establish a migration model ,then calculate migrated cells after 48hours. Resuls There were no difference between normal control group and empty plasmid group in cells grow, MMP-2’s content , TIMP-3 mRNA expression and apoptosis rate,cells migration rate.While recombinant group had obvious difference compared with the other two,P<0.05. Conclusions It is testified TIMP-3 can strongly inhibit proliferation or migration of VSMC in vitro, but promote apoptosis on the other hand. Cationic liposome is a safe medium which can provide broad application in TIMP-3 gene therapy on ISR.

Key words: TIMP-3, vascular smooth muscle cells, cationic liposome, gene transfection, in-stent restenosis