【摘要】 目的　将携带人血管内皮生长因子 16 5 (hVEGF 16 5 )cDNA的重组腺病毒相关病毒 (rAAV)载体注入家兔缺血心肌 ,观察血管内皮生长因子 (VEGF)的血管生成效应。方法　制作家兔急性心肌梗死模型 ,将rAAV hVEGF 16 5注入缺血心肌。 4周后取注射部位心肌 ,用逆转录聚合酶链式反应 (RT PCR)和免疫组织化学、Westernblot方法检测目的基因的表达 ,并计数注射部位心肌毛细血管数目 ,评价外源VEGF的生物学活性。结果　注射rAAV VEGF部位心肌VEGFmRNA及其蛋白的表达可持续 8周 ,而且比对照组增加。远隔脏器未见外源VEGF的播散和表达。注射rAAV VEGF部位毛细血管密度比对照组增加。结论　rAAV hVEGF 16 5可以有效地转染成年家兔心肌组织 ,外源基因稳定长期表达 ,同时能够刺激新血管生成 ,并具有良好的安全性。更多还原

【Abstract】 Objective To investigate the therapeutic potential of recombinant adeno-associated virus mediated vascular endothelium growth factor (rAAV-hVEGF?165) gene for ischemic heart diseases.Methods A rabbit myocardial ischemic model was formed by ligation of the anterior descending coronary artery. and rAAV-hVEGF?165?cDNA was injected into the ischemic myocardium. Gene expression was evaluated by RT-PCR,immunohisto-chemical (IHC) analysis and Western blotting. Myocardial capillary density was calculated to show the biological activity of expressed VEGF protein.Results VEGF mRNA and protein expression was consistently observed in the injected sites for at least 8 weeks after the injection, while no VEGF mRNA could be detected in remote organs. Capillary density was significantly higher in rAAV-VEGF-injected tissues than in PBS-injected tissues.Conclusions This study demonstrates that AAV-mediated VEGF gene transfer into rabbit ischemic heart is efficient and stable without ectopic expression, and AAV-mediated VEGF gene transfer stimulates angiogenesis.These findings suggest that AAV-mediated VEGF gene transfer may be useful for treatment of ischemic heart diseases.更多还原