组织因子途径抑制物基因转移对动脉血栓形成的抑制作用

Inhibition of Tissue Factor Pathway Inhibitor Gene Transfer on the Arterial Thrombus Formation

目的:研究日本凝血病毒(HVJ)-人工病毒包膜(AVE)脂质体介导的组织因子途径抑制物(TFPI)基因转移对损伤血管局部血栓形成抑制作用的有效性及安全性。方法:在兔球囊损伤髂动脉局部分别转染含TFPI基因(TFPI组)或pcDNA3．1质粒(空质粒组)的HVJ-AVE脂质体复合物或HVJ-脂质体(空载体组)或生理盐水(盐水组),1,3和7天后用逆转录多聚酶链反应和免疫组化法检测血管局部TFPI表达;组织病理学和扫描电镜观察比较各组血栓形成情况;各组实验动物观察6个月评价这种疗法的安全性。结果:TFPI基因转移后1天即检测到TFPI mRNA表达,第3天为高峰;TFPI组血栓形成例数明显少于其它3组(P<0.05);体内凝血指标及生化指标无明显变化;重要脏器组织病理学检查未见明显变化;各器官未见到HVJ感染及复制迹象。结论:HVJ-AVE脂质体介导TFPI基因转移能够安全而有效地抑制损伤血管局部血栓形成。

Objective ： To investigate the validity and feasibility of the inhibition of hemagglutinating virus of Japan （ HVJ ） -artificial viral envelope（AVE） liposome mediated tissue factor pathway inhibltor（TFPI） gene transfer on thrombosis of injured vessels. Methods： local iliac artery injury was induced by balloon denudation. Infusion of an HVJ -AVE liposome containing TFPI gene （TFPI group）or empty pcDNA3. 1 plasmid （empty plasmid group） or HVJ-liposome vector （empty vector group） or saline （saline group） was performed at the sites of injured vessels. The exogenous TFPI gene expression in injured arteries was detected by reverse transcription-polymerase chain reaction and immuno-histochemical method. Thrombus formation in each group was observed by histopathological examination and electron microscopy. The safety of gene therapy was evaluated after a six-month obserbation. Results ： The expression of TFPI mRNA was detectable at the first day after gene transfer and the peak took place at the third day. The thrombosis was significantly reduced in TFPI group compared with the other three groups（ P 〈 0. 05）. The systemic coagulation and biochemistry indexes had no obvious changes between the 4 groups. Pathological findings in major organs showed no obvious changes. No HVJ infection or replication were found after HVJ-AVE liposome-mediated gene transfer. Conclusion： HVJ-liposome mediated TFPI gene transfer can effectively and safely inhibit thrombosis in local injured vessels.