血管内皮生长因子基因治疗家兔肢体缺血性疾病的实验研究

应用血管内皮生长因子 (VEGF) ,对肢体缺血性疾病进行基因治疗的实验性研究 ,同时寻找适用于临床的、有效衡量肢体血液循环状况的检测指标。应用股动脉及其分支分离剔除术 ,造成新西兰兔下肢闭塞性血管病模型 ,术后立即于缺血骨骼肌部位多点注射 pcDNA3 .1 /VEGF16 5质粒 (n =1 2只 ) ,pcDNA3 .1空质粒 (n =8只 ) ,生理盐水(n =2 2只 )。选取不同时间点 ,行颈动脉插管造影 ,测量胫动脉压 ,检测双后肢膝关节的单位时间血流量、血流速度和红细胞浓度 ,检测肝肾功能 ,并进行病理取材 ,观察病理变化。结果显示 :术后对照组出现肢体缺血性坏死的动物数明显多于基因治疗组 ;血管造影显示 ,术后基因治疗组远端动脉充盈早于对照组 ,新生血管数目也明显多于同时期对照组 ;VEGF基因治疗组术后动脉血压的恢复也较对照组快。病理切片观察对照组骨骼肌出现萎缩坏死、间质纤维组织增生等缺血性病理改变的程度较基因治疗组重 ,质粒组新生血管明显多于对照组。结果提示 :利用肌肉细胞能够自主摄取裸DNA的特性 ,用重组VEGF质粒进行基因治疗 ,有效改善了缺血肢体的血液供应 ,并未出现严重不良反应

Treatment of Limb Ischemia in Rabbits by VEGF Gene

The objective was to study the gene therapy effects of vascular endothelial growth factor(VEGF) and to find the new approaches to describe the development and evaluation of blood circulation on a rabbit hindlimb ischemic model. The limb ischemic model was developed by ligation of the distal external iliac artery and excision of every branches of femoral arteries. As soon as the model was established, the recombinant plasmids pcDNA3.1/VEGF165 was transferred by injection into the ischemia muscles. Each animal was evaluated by angiography, calf blood pressure, blood volume of keen per second, speed of blood circulation and erythrocyte concentration per second on keens of the rabbit. At the same time the histologic changes of the adductor muscle and the gastrocnemucs muscle in the left and right hindlimbs were studied. In control group the number of the rabbits which developed superficial tissue necrosis in the foot was greater than those in experimental group. Angiography demonstrated that in the experimental group, the filling of the distal arterial was markedly faster than in the control group and focal neovessels establishment of collateral circulation was higher than that in the control group at the same time. The calf blood presure in the experimental group increased faster than that in the control group. Histologic study showed that the evidences of atrophy and fibrosis in the control group were more obvious than those in the experimental group and neovessels were less than the experimental group. Between the two groups there were not significant difference in the function of liver and kidney. The experimental studies indicate that VEGF gene therapy would be effective and safe for limb ischemia diseases.