人TFPI基因转染对兔移植静脉血栓形成和近期通畅率的影响

目的： 为减少冠状动脉旁路移植术后血栓形成，探讨人组织因子途径抑制因子(TFPI)基因转染对兔移植静脉血栓生成的影响。方法：构建真核表达质粒pCMV-(Kozak)TFPI。采用阳离子脂质体和腔内加压灌注法，转染移植静脉内皮细胞。RT-PCR、Western blotting和免疫组化法检测外源基因在兔移植静脉中的表达。病理标本、扫描电镜观察移植静脉血栓形成情况，血管多普勒观测其通畅率。结果：移植静脉中有人TFPI基因和蛋白表达。静脉移植术后3 d，基因转染组、空载体和空白对照组分别有1条、8条和7条移植静脉发生血栓，术后30 d，以上各组分别有0条、5条和5条移植静脉完全闭塞，前者静脉血栓发生率和闭塞率均低于后两者(P<0.05)。扫描电镜显示两对照组内皮表面有红细胞和血小板黏附、聚集，而转染组基本正常。结论：人TFPI基因干预，减少了兔移植静脉血栓形成，提高了近期通畅率。

Effect of human tissue factor pathway inhibitor gene transfection in vein grafts on thrombus formation and early patency rate

AIM: To reduce thrombus formation after coronary artery bypass graft，we investigated the antithrombotic effect of human tissue factor pathway inhibitor (TFPI) gene delivery on vein grafts. METHODS: The eukaryotic expressed plasmid vector pCMV-(Kozak)TFPI was constructed. Through pressurizing infusion，vein endotheliocytes were transfected with cationic liposome containing the plasmid vector pCMV-(Kozak)TFPI. After operation, vein grafts were harvested at third day for immunohistochemical, RT-PCR and Western blotting analysis of exogenous gene expression and for observation of thrombus formation by pathological method and scanning electron microscopy. At 30th days, the patency rate was recorded by vessel Doppler ultrasonography. RESULTS: Human TFPI mRNA and protein were detected in TFPI gene transferred vein grafts. Thrombosis was found in 8 animals of empty plasmid control group and in 7 animals of empty control group, but in only 1 of the TFPI group (P<0.05). Thirty days after operation, 5 vein grafts were occluded in both empty plasmid control group and empty control group, but none of vein grafts were occluded in TFPI group (P<0.05). The endothelial surfaces of the vein grafts in both control groups were covered with aggregated erythrocytes and platelets, but not in TFPI group. CONCLUSION: Human tissue factor pathway inhibitor gene transfection reduces thrombus formation and improves early patency rate in vein grafts.