组织因子途径抑制物基因局部转染对兔颈总动脉损伤后内膜增生及P53表达的影响

Local Transfection of Tissue Factor Pathway Inhibitor Gene on Neointimal Proliferation and Protein-53 Expression in Rabbit Carotid Artery After Balloon Injury

目的研究组织因子途径抑制物(TFPI)基因局部转染对兔颈总动脉损伤处内膜增生及P53表达的影响。方法36只新西兰纯种雄性大白兔颈总动脉损伤后随机分生理盐水组?复制缺陷型腺病毒(AdLacz)组和包载TFPI目的基因复制缺陷型腺病毒(AdTFPI)组。每组12只。术后14d处死动物,取损伤血管进行检测分析。免疫组化染色观察TFPI在转染局部的表达成功;颈总动脉损伤处抽提总RNA,半定量逆转录多聚酶链式反应(RT-PCR)测定P53mRNA表达;同时对颈总动脉病变最明显的取材标本进行HE染色,光学显微镜下观察血管内膜增生,以计算机图像分析系统分析血管内膜?中膜面积和腔面积。结果AdTFPI组家兔的颈总动脉损伤处血管内膜的增生显著受抑制?内膜/中膜面积比值下降;生理盐水组?AdLacz组和AdTFPI组P53mRNA的相对值分别为1.31±0.14、1.27±0.11和4.21±0.87,P53在AdTFPI组表达增强,与前两者相比有显著差异性。结论TFPI基因局部转染显著抑制颈总动脉损伤处血管内膜的增生并增强P53表达。

Objective To assess influence of local transfection of tissue factor pathway inhibitor gene on neointimal hyperplasia and protein-53（P53） expression in rabbit carotid artery after balloon injury. Methods 36 male New Zealand white rabbits were divided into three groups： saline, adenovirus-defect and adenovirus-defect carrying tissue factor pathway inhibitor gene treated group. Balloon catheter was used to induce injury to the carotid artery of rabbits and saline, adenovirus-defect and TFPI gene were locally delivered to injury site. 14 days after procedures, the local injury vesseles were removed and harvested for pathological and immunohistochemical analysis. Expression of protein-53 was determined by using serniquantitive RT-PCR . Automarie image analysis system was used to detect the changes of neointimal , medium and luminal area. Results TFPI transfection could remarkably inhibit neointimal proliferation. The relative ratio of P53 mRNA was 1.31 ± 0.14, 1.27 ± 0. 11 and 4.21 ± 0.87 in saline, adenovirus-defect and TFPI local transferring respectively. Comparied with saline and adenovius defect group, TFPI local transferring obviously increased P53 expression. Conclusion Local TFPI gene transfection can promote P53 expression and reduce neointimal hyperplasia in carotid artery of rabbits.