摘要
目的验证血管内皮生长因子（ＶＥＧＦ）基因治疗肢体动脉梗塞病的有效性和安全性。方法构建重组ＶＥＧＦ基因的真核表达载体ｐｃＤ２／ＶＥＧＦ。有效性试验：制备肢体动脉梗塞病的猴模型，通过肌肉基因缝线法和肌内多点注射法导入ＶＥＧＦ基因，对照组转等量空载质粒。安全性试验：将超剂量和治疗剂量的ＶＥＧＦ基因分别导入５只小鼠（１０００μｇ，为小鼠治疗剂量２０μｇ的５０倍）、１０只大鼠（２０００μｇ，为治疗剂量的１０倍）和５只猴（２０００μｇ，治疗剂量）体内观察其毒副作用，并采用ＰＣＲ法和间接ＥＬＩＳＡ法进行分子遗传学和免疫学的评估。结果转ＶＥＧＦ基因后１周血管造影即显示，结扎点附近有较多充盈小血管，远端动脉开始充盈，２周时更为明显，１月后新生小血管最为丰富，可见远端动脉明显充盈，血管计数显示ＶＥＧＦ组（１周时５／１００ｃｍ２、１月１２／１００ｃｍ２）明显多于对照组（１周１／１００ｃｍ２、１月４／１００ｃｍ２）。生化等检查，证明ＶＥＧＦ基因对肝肾功能等无明显影响，未见肿瘤或其它病理变化。导入的ＶＥＧＦ基因可在局部肌肉组织中保存４个月以上，不累及其它各脏器组织，且无循环抗体产生。结论肌肉导入ＶＥＧＦ基因治疗肢体动脉梗塞病是有效和?
Objective To Study experimentally VEGF gene for treatment of peripheral artery disease.Methods The human VEGF cDNA was cloned into the eukaryotic expression vector pcD 2.Using gene suture, the recombinant plasmid was transferred into the hindlimbs′ adductor of Rhesus monkey, of which the distal end of the external iliac arteries were ligated and the femoral arteries were completely excised. With angiography, the biological effect of VEGF gene in experimental animals was investigated. Safey tests were analyzed by transferring of VEGF into mouse, rat, and Rhesus monkey, evaluated by biochemistry analysis, histopathological examination, PCR, and indirect ELISA.Results The transfer of VEGF gene stimulated the formation of focal microvessels, established collateral circulation, and augmented blood perfusion. The system had no adverse effect and remarkable pathological change in mouse, rat, and Rhesus monkey.Conclusion The experimental studies indicated that VEGF would be effective and safe for clinical trial after approval.
出处
《中华医学杂志》
CAS
CSCD
北大核心
1999年第2期129-132,共4页
National Medical Journal of China
基金
国家"八六三"计划生物技术资助
关键词
肢体动脉梗塞病
基因治疗
内皮生长因子
Gene therapy Endothelial growth factor Peripheral vascular diseases
