联合转染VEGF和PCNA-ASODN对血管成形术后再狭窄的影响

Effect of co-transfection of VEGF and PCNA-ASODN on restenosis after percutaneous transluminal coronary angioplasty

目的探讨联合转染血管内皮生长因子(VEGF)基因和增殖细胞核抗原反义寡核苷酸(PCNA-ASODN)对血管成形术后再狭窄的影响。方法 32只新西兰纯种大白兔随机平均分为对照组、转染VEGF组、转染PCNA-ASODN组、联合转染VEGF和PCNA-ASODN组(联合转染组)。构建兔的股髂动脉球囊损伤术后再狭窄模型,以医用生物蛋白胶作为缓释载体,将300μg PCNA-ASODN或VEGF均匀喷布于处理血管段外膜。取损伤处理血管段标本,光学显微镜观察损伤血管内膜、中层结构和局部血栓形成情况;电子显微镜观察新生内膜内皮细胞及内膜、中膜,计算内膜/中膜(I/M)面积比与厚度比;采用RT-PCR法检测VEGF与PCNA基因的表达,采用免疫组化从蛋白质水平检测VEGF与PCNA的表达。结果联合转染组与其他组相比,其血管内皮较完整、光滑,内膜轻度增生,平滑肌细胞排列规则,外膜无明显变化;I/M面积比与厚度比明显较低(P<0.05);RT-PCR检测VEGF表达水平明显升高(P<0.05);免疫组化检测血管组织中VEGF蛋白表达阳性率较高(P<0.05)。结论联合转染VEGF和PCNA-ASODN能更有效地抑制球囊损伤术后的血管内膜增生和再狭窄的发生。

Objective To explore the effect of co-transfection of vascular endothelial growth factor（VEGF） and proliferating-cell-nuclear-antisense-antigen oligodeoxynucleotides（PCNA-ASODN） on restenosis after percutaneous transluminal coronary angioplasty.Methods 32 Zelanian rabbits were randomly divided into four groups： the control group, the VEGF group,the PCNA-ASODN group,and the VEGF＋PCNA-ASODN group.Using pluronic gel as the control-released carrier, 300 μg of VEGF or PCNA-ASODN were given to the adventitia of the femoral iliac artery with restenosis after balloon injury in rabbits. The intima, middle structure and local thrombosis in the injured vascular specimen were observed by an optical microscope,and neointimal endothelial cells, the intima and media were examined by an electronic microscope. The ratio of intima/media（I/M） areas and the ratio of I/M thickness were determined.mRNA expressions of VEGF and PCNA were measured by reverse transcription-PCR. Protein expressions of VEGF and PCNA were determined with immunohistochemistry. Results Compared with the other groups, vascular endothelium in the VEGF＋PCNA-ASODN group was intact, smooth and mildly hyperplasic, and vascular smooth muscle cells were arranged in order,while there was no obvious change in the adventitia.Ratios of I/M area and thickness were significantlylower （P〈0.05）. mRNA expression of VEGF was significantly increased by RT-PCR（P〈0.05）. Protein expression of VEGF was significantly higher by immunohistochemistry（P〈0.05）. Conclusion Co-transfection of VEGF and PCNA-ASODN could effectively inhibit neointimal hyperplasia and restenosis after balloon injury.