组织因子途径抑制物基因对支架内再狭窄的抑制作用

The Effect of Adenovirus-Mediated Local Human Tissue Factor Pathway Inhibitor Gene on In-Stent Restenosis

目的探讨腺病毒介导的人组织因子途径抑制物基因转移抑制兔髂股动脉支架内再狭窄的有效性、安全性及相关机制。方法对36只新西兰白兔进行双侧髂股动脉球囊损伤,然后植入镍钛合金自膨胀支架,再分别局部灌注保留腺病毒介导的人组织因子途径抑制物基因、细菌β半乳糖苷酶基因和生理盐水。术后第3、7、28天分别取各组实验血管段,用X-Gal染色观察外源基因的转染效率,逆转录聚合酶链反应检测组织因子途径抑制物基因mRNA的表达,免疫组织化学染色检测血管壁细胞增殖细胞核抗原的表达情况,细胞凋亡原位检测法确定血管壁细胞的凋亡情况,组织形态学观察研究管腔的狭窄程度,血液生物化学及组织形态学评价基因治疗的安全性。结果外源组织因子途径抑制物基因成功转染至血管壁细胞并表达成mRNA;组织因子途径抑制物可抑制平滑肌细胞的增殖(P<0.05)、促进其凋亡(P<0.05),抑制支架内再狭窄形成(P<0.05),各组动物重要脏器无病理学改变。结论组织因子途径抑制物基因有效抑制了兔髂动脉支架内再狭窄的形成且无明显毒副作用,其机制与抑制平滑肌细胞增殖及促进其凋亡有关。

Aim To investigate the effect, safety and mechanism of adenovirus-mediated local human tissue factor path-way inhibitor （TFPI） gene on in-stent restenosis. Methods The ilio-femoral arteries of thirty six New Zealand white rabbits were inflated by balloon angioplasty catheter and implanted nickel titanium alloy self-expended stents. Adenovirus-mediated TFPI gene （Ad-TFPI）, bacterial β-galactosidase gene （Ad-LacZ） and saline were irrigated for 20 minutes at the site of stents respectively. At the day of 3, 7 and 28 after injuried, experimental arteries were harvested and studied the targets such as the exogenous gene transfer rate and expression were studied, and immunohistochemistry staining （proliferating cell nuclear antigen, PC- NA）, TUNEL assay （terminal dUTP nick-end labeling to detect apoptotic cells）, histomorphometry and blood biochemical detection were used. Results Genes were transferred and expressed successfully. Compared with Ad-LacZ and saline groups, Ad-TFPI group＇s vascular smooth muscle cell （VSMC） proliferation rate was significantly decreased （ P 〈 0.05 ） and VSMC apoptosis rate was significantly increased on the 7th day （ P 〈 0.05）. The mean neointimal area, the ratio of the neointimal to medial areas, and percent of stenosis in the TFPI group were all significantly reduced compared with the control groups （ P 〈 0.05）. The main organs and blood biochemical indicators of the treated animals were not changed in all groups. Conclusions Ad-TFPI gene transfer significantly reduced neointimal hyperplasia, suppressed cell proliferation, induced cell apoptosis without systemic side effects.