Feng Deguang, Li Quan, Zhang Kailun, Jiang Xionggang, Leng Song, Deng Heping, Feng Jian'e, Sun Tucheng, Wu Long, Zhou Cheng
Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2008 Apr;28(2):147-51. doi: 10.1007/s11596-008-0208-4. Epub 2008 May 15.
To develop a more efficient antithrombotic way after coronary artery bypass grafting (CABG), the anticoagulant effects were compared of human tissue factor pathway inhibitor (TFPI) gene transfection and aspirin oral administration (traditional method) on vein grafts. An eukaryotic expression plasmid pCMV-(Kozak) TFPI was prepared. Animal model of carotid artery bypass grafting was constructed. In operation, endothelial cells of vein grafts in TFPI group and empty plasmid control group were transfected with pCMV-(Kozak) TFPI and empty plasmid pCMV respectively, while no transfection was conducted in aspirin control group. After operation, aspirin (2 mg.kg(-1).(-1)) was administered (i.g.) in aspirin control group. Three days later, grafts (n=10) were harvested for RT-PCR, Western blotting and immunohistochemical analyses of exogenous gene expression and for pathological, scanning electron microscopic observation of thrombus. Thirty days later, the patency rates of remnant grafts (n=10) were recorded by vessel Doppler ultrasonography. Human TFPI gene products were detected in gene transferred vein grafts. Three days later, thrombi were found in 7 animals of aspirin control group and in 8 animals of empty plasmid control group, but in only 1 of TFPI group (P<0.01). Thirty days later, 5 grafts were occluded in empty plasmid control group, but none of grafts was occluded in the other groups (P<0.05). The endothelial surfaces of grafts in both of the control groups were covered with aggregated erythrocytes and platelets, and it were not seen in TFPI group. It was suggested that the anticoagulant effects on vein grafts of human TFPI gene transfection are better than those of aspirin.
为了在冠状动脉旁路移植术(CABG)后开发一种更有效的抗血栓形成方法,比较了人组织因子途径抑制剂(TFPI)基因转染和口服阿司匹林(传统方法)对静脉移植物的抗凝作用。制备了真核表达质粒pCMV-(Kozak)TFPI。构建了颈动脉旁路移植动物模型。手术中,TFPI组和空质粒对照组的静脉移植物内皮细胞分别用pCMV-(Kozak)TFPI和空质粒pCMV转染,而阿司匹林对照组未进行转染。术后,阿司匹林对照组经灌胃给予阿司匹林(2mg·kg⁻¹·d⁻¹)。三天后,采集移植物(n = 10)进行RT-PCR、Western印迹和免疫组织化学分析以检测外源基因表达,并进行病理、扫描电子显微镜观察血栓。三十天后,通过血管多普勒超声记录残余移植物(n = 10)的通畅率。在基因转染的静脉移植物中检测到了人TFPI基因产物。三天后,阿司匹林对照组7只动物和空质粒对照组8只动物发现有血栓形成,而TFPI组仅1只动物出现血栓(P<0.01)。三十天后,空质粒对照组有5只移植物闭塞,而其他组无移植物闭塞(P<0.05)。两个对照组移植物的内皮表面均覆盖有聚集的红细胞和血小板,而TFPI组未见此现象。提示人TFPI基因转染对静脉移植物的抗凝作用优于阿司匹林。