重组腺相关病毒载体介导人血管内皮生长因子165基因对缺血心肌血管新生的影响

为了研究重组 2型腺相关病毒载体介导人血管内皮生长因子 16 5 (hVEGF16 5 )基因对兔缺血心肌血管新生的影响 ,选取了 4 0只新西兰兔 ,建立心肌缺血模型后随机分为血管内皮生长因子高、中、低剂量组及对照组等 4组 ,分别向缺血区域心肌注射不同剂量的腺相关病毒载体 人血管内皮生长因子 16 5或磷酸盐缓冲液。 4周后取心肌组织和血液标本 ,采用逆转录聚合酶链反应和酶联免疫吸附法检测人血管内皮生长因子 16 5基因的表达 ;制作组织学切片以观察心肌组织的病理改变 ,于高倍镜下计数缺血区域毛细血管数目。结果发现 ,血管内皮生长因子低、中、高剂量组的人血管内皮生长因子 16 5 GAPDHmRNA比值依次为 0 .14± 0 .0 3、0 .4 0± 0 .0 4和 0 .6 4± 0 .0 4 ,血清人血管内皮生长因子 16 5含量分别为 6 4 .6± 8.0ng L、32 7.1± 9.9ng L和 4 71.6± 6 .9ng L ,各组间的差异均具有显著性 (P <0 .0 1)。低、中、高剂量组单个高倍视野内的毛细血管数分别为 4 .6± 1.3、11.6± 1.8和 2 1.8± 3.1条 ,而对照组为 4 .5± 1.5条。高、中剂量组的毛细血管数显著高于对照组 (P <0 .0 1) ,而低剂量组与对照组相比差异无显著性 (P >0 .0 5 )。相关分析发现毛细血管数与腺相关病毒载体 人血管内皮生长因子 16 5剂量呈

Recombinant Adeno-associated Viral Vector2-Mediated Human Vascular Endothelial Growth factor 165 Gene Transfer Induces Angiogenesis in Ischemic Heart

Aim To investigate the effect of human vascular endothelial growth factor 165 (hVEGF165) cDNA mediated by recombinant adeno-associated viral vector2 (rAAV-2) on the angiogenesis in ischemic myocardium. Methods Rabbit myocardial ischemic models were generated by ligation of the anterior descending coronary artery. All these models were randomly divided into 4 groups: VEGF high dose group (10 12v.g./kg), VEGF middle dose group (10 11v.g./kg), VEGF low dose group (10 10v.g./kg) and control group. RAAV-2/hVEGF165 or PBS was injected into the ischemic myocardium respectively. After 4 weeks, the expression of objective gene was evaluated by RT-PCR and ELISA. The histological changes of myocardium were observed through histological sections. Myocardial capillary counts were calculated to evaluate the proangiogenic effects. Results With the increasing dosage of injected rAAV-2/hVEGF165, hVEGF165/GAPDH mRNA ratio were 0.14±0.03, 0.40±0.04 and 0.64±0.04 respectively, while VEGF protein content were 64.6±8.0 ng/L, 327.1±9.9 ng/L and 471.6±6.9 ng/L respectively. The changes of expression were dose-dependent and there were significant difference among the groups(p<0.01). In rAAV-2/hVEGF165 10 10, 10 11 and 10 12 v.g./kg treatmented groups, the microvessel counts were (4.6±1.3)/HPF, (11.6±1.8)/HPF and (21.8±3.1)/HPF respectively, while (4.5±1.5)/HPF in control group. There was significant difference between the 10 11, 10 12 v.g./kg hVEGF165 gene-treated groups and control group(both p<0.01), but no such difference between 10 10 v.g./kg hVEGF165 gene-treated group and control group(p>0.05). The result of correlation analysis showed the microvessel counts were related with the dosages of rAAV-2/hVEGF165 positively (r=0.910,p<0.01). Conclusions hVEGF165 gene mediated by rAAV-2 can be efficiently transferred into rabbit ischemic heart and induce angiogenesis of myocard-