超声定位辐照载pEGFP质粒DNA超声微泡转染新生血管性角膜组织的实验研究  

Transferring pEGFP gene into the neovascularized corneal mediated by ultrasound microbubbles

作　　者：胡文静[1] 周善璧[1] 王志刚[2] 许川山[2] 

机构地区：[1]重庆医科大学附属第一医院眼科,400016 [2]重庆医科大学超声影像学研究所

出　　处：《中华超声影像学杂志》2009年第8期722-725,共4页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金资助项目（30430230）;国家高技术研究发展计划（“863”计划）资助项目（2006AA02Z4F0）志谢本实验在重庆医科大学超声影像学研究所、重庆医科大学附属第一医院中心实验室支持下完成,特此致谢

摘　　要：目的探讨超声微泡介导pEGFP质粒DNA转染新生血管性角膜组织的可行性及效率。方法24只新西兰大白兔采用缝线法诱导角膜新生血管成功后，利用随机分组法将其分为A，B，C，D4组（每组6只）：A组为裸质粒组；B组为质粒加超声辐照组；C组为质粒-微泡复合物组；D组为质粒-微泡复合物加超声辐照组。质粒转染后HE染色观察角膜组织病理变化，并采用激光共聚焦显微镜观察质粒DNA荧光蛋白表达强度。结果超声破坏载pEGFP质粒的微泡，D组荧光蛋白表达强度（133．03±21．99）明显高于其他3组（A组：75．17±7．08；B组：97．02±8．31；C组：81．83±9．44），差异有统计学意义（F=23．56，P〈0．05）。结论一定声强和时间的辐照下，超声微泡能有效提高质粒DNA在兔新生血管性角膜组织的基因转染率。Objective To explore whether ultrasound （US） microbubble （MB） could directional deliver （plamid vector coding enhanced green fluorescence pro tein,pEGFP） into the neovascularized corneal of rabbit and definite gene transfect efficiency. Methods After use suture induced corneal angiogenesis, twenty-four New Zealand albino rabbits were randomly divided into four groups：Group A were the naked plasmid DNA of pEGFP into rabbit subconjunctival;Group B were taken ultrasound（US） irradiation after injected pEGFP immediately ; The plasmid DNA of pEGFP and microbubbles（MB） complex were injected into rabbit subconjunetival in group C; Group D were injected the complex（plasmid ＋ MB） and then took US immediately. The results were observed by histopathologic and Leica confocal microscope. Results Group D greatly enhance plasmid DNA of pEGFP expressed in cornea, the ratio of EGFP positive was 133.03 ± 21.99, and the three other groups were 75.17 ±7.08;97.02 8.31 and 81.83 ± 9.44, respectively（ F = 23.56, P 〈0.05）. Conclusions Limited ultrasound energy can obviously improve plasmid DNA of pEGFP transfect efficiency in neovascularized corneal of rabbit.

关 键 词：超声检查 微气泡 角膜新生血管化 转染 

分 类 号：R686[医药卫生—骨科学]