组织因子途径抑制物基因转移对血管平滑肌细胞迁移的影响  

Effect of Tissue Factor Pathway Inhibitor Gene on the Migration of Vascular Smooth Muscle Cell

作　　者：傅羽[1] 尹新华[1] 张一娜[2] 李呼伦[3] 周令望[4] 

机构地区：[1]哈尔滨医科大学附属第二医院心血管内科 [2]哈尔滨医科大学附属第二医院老年病科 [3]哈尔滨医科大学神经生物学教研室,黑龙江省哈尔滨市150086 [4]哈尔滨医科大学克山病研究所,黑龙江省哈尔滨市150086

出　　处：《中国动脉硬化杂志》2006年第2期111-114,共4页Chinese Journal of Arteriosclerosis

基　　金：黑龙江省自然科学基金(D00-20);黑龙江省教育厅海外学人资助(1053HQ021)

摘　　要：目的研究组织因子途径抑制物基因转移对兔血管平滑肌细胞迁移的影响,探讨其抑制再狭窄的机制。方法将含有人组织因子途径抑制物基因的重组腺病毒和含β-半乳糖苷酶基因的重组腺病毒在体外分别转染兔血管平滑肌细胞,用X-gal染色法检测腺病毒的转染率,用逆转录聚合酶链反应方法检测转染后平滑肌细胞中组织因子途径抑制物mRNA的表达,于第13、、5及7天用玻片法检测血管平滑肌细胞迁移距离。结果感染指数为100,腺病毒的转染率可达到95%;基因转移后3天在血管平滑肌细胞中检测到组织因子途径抑制物mRNA的表达;同一浓度下转染组织因子途径抑制物基因重组腺病毒组和对照组相比迁移距离显著减少(P<0.001),并具有浓度依赖性。结论腺病毒具有较高的转染培养的血管平滑肌细胞的能力,组织因子途径抑制物基因转移能够显著抑制体外培养的兔血管平滑肌细胞迁移,并且这种抑制作用具有浓度依赖性。Aim To investigate the effect of tissue factor pathway inhibitor （TFPI） gene transfer on the migration of vascular smooth muscle cell and the mechanism of its inhibition on the restenosis. Methods Human tissue factor pathway inhibitor recombinant adenovirus and LacZ recombinant adenovirus were used to infect rabbit aortic vascular smooth muscle cell respectively in vitro. The transfection efficiency was examined by X-gal staining. The exogenous tissue factor pathway inhibitor gene expression in transfected cell was detected by reverse transcription-polymerase chain reaction （RT-PCR） and its effect on migration was tested with slide method. Results When multiplicity of infection was 100, the transfection efficiency was 95 percent. Expression of tissue factor pathway inldbitor mRNA was detected in TFPI gene transfected vascular smooth muscle cell at 3rd day after gene transfer. At the same concentration, the migration distances were remarkably reduced in tissue factor pathway inhibitor gene transfected groups compared with the control group （ P 〈 0. 001 ）, and had a dependence of concentrations. Conclusions Adenovirus has a higher transfection efficiency on the cultured vascular smooth muscle cell. Tissue factor pathway inhibitor gene transfer mediated by adenovirus can greatly inhibit the migration of cultured rabbit vascular smooth muscle cell and have a dependence of concentration.

关 键 词：内科学 血管成形术后再狭窄的基因治疗 逆转录聚合酶链反应 组织因子途径抑制物 血管平滑肌细胞 基因转移 细胞培养 

分 类 号：R5[医药卫生—内科学]