hVEGF_(165)对下肢股浅静脉非体外循环下行冠状动脉旁路移植静脉术血管吻合口再狭窄的影响

Effect of hVEGF_(165) transfection to prevent vascular anastomotic restenosis after coronary aorta bypass grafting under superficial femoral vein in off-pump

目的观察应用血管内皮细胞生长因子165(hVEGF165)基因治疗狗冠状动脉-主动脉旁路移植术吻合口再狭窄的疗效。方法构建pcDNA3/VEGF165重组质粒转染中国仓鼠卵巢细胞Kl亚株(CHO-K1),检测其基因、蛋白表达并转导冠状动脉架桥静脉移植术吻合口局部血管平滑肌细胞,观察其对吻合口内膜厚度、面积、狭窄率、新生内膜心肌血管内皮细胞(MEC)增殖及增殖细胞核抗原(PCNA)的影响。结果质粒构建及转染成功,pcDNA3/VEGF165能显著促进MEC增殖,并减少狗冠状动脉-主动脉旁路移植术吻合口再狭窄模型再狭窄血管的内膜厚度、面积、狭窄率。结论 VEGF165重组质粒可有效改善狗冠状动脉-主动脉旁路移植术吻合口再狭窄,为临床应用提供了前期研究基础。

Objective To elucidate the transfection of vascular endothelial growth factor 165（VEGF165） gene to prevent vascular anastomotic restenosis after coronary aorta bypass grafting.Methods pcDNA3/VEGF165 recombinant plasmid was constructed and its gene thread was made to transfect the Chinese hamster ovary cell sublines Kl （CHO-K1）, cell VEGF165 gene expression and VEGF165 protein expression from cell supernatant was detected. After pcDNA3/VEGF165 plasmid and myocardial vascular endothelial cells （MEC） and proliferating cell nuclear antigen（PCNA） co-cultured, changes in endothelial cell proliferation force were detected. Dog coronary restenosis-aortic bypass graft anastomosis model was constructed, anastomosis was treated by pcDNA3/VEGF165 plasmid,then intimal thickness, area, narrow rate of anastomotic and neointimal proliferation was observed. Results pcDNA3/VEGF165 plasmid was successful constructed and transfected to CHO-K1 cell lines, VEGF165mRNA in cell lines and VEGF165 protein from supernatant significantly increased after transfection. pcDNA3/VEGF165 plasmid can significantly promote the MEC cell proliferation and significantly reduce the dog coronary artery - the aorta bypass graft anastomotic restenosis model restenosis vascular intimal thickness, area, the rate of stenosis and intimal cell proliferation. Conclusion VEGF165 recombinant plasmid can inhibit the anastomotic restenosis of the aorta-coronary artery bypass graft in dog.