P123-PPI%2fDNA纳米粒性质表征与体外转染效率研究.pdfVIP

P123-PPI/DNA 纳米粒性质表征和体外转染效率研究 郝俊国，沙先谊，蒋晔，方晓玲* （复旦大学药学院药剂教研室，上海200032 ） 摘要 目的 研究开发一种高效、安全的非病毒基因给药载体系统；方法 合成 P123 修饰的 聚丙烯亚胺 （polypropyleneimine ，PPI ）树状高分子，对P123-PPI/P123 压缩DNA 形成的 纳米粒的粒径和 ? 电位进行表征，以琼脂糖凝胶电泳实验考察纳米粒抵抗血清和肝素钠降 解的作用和纳米粒保护质粒 DNA 不受DNaseI 降解的作用，以绿色荧光蛋白基因质粒作为 报告基因，考察 P123-PPI/P123 载体材料的体外转染效率；结果 P123-PPI/P123 压缩 DNA 形成的纳米粒的粒径在 100nm 左右，? 电位约 15mV，纳米粒能够抵抗血清和一定浓度肝 素钠的解离作用，P123-PPI/P123 具有较高的体外转染效率，显著高于PPI 本身；结论 P123 修饰PPI 具有较高的体外转染效率，有可能开发成一种具有较高体内转染效率的载体系统。 关键词：基因治疗；Pluronic P123 ；聚丙烯亚胺树状告分子 中图分类号： 文献标识码： 文章编号： Characterization of Nanoparticles Formed with P123-PPI and Plasmid DNA and Study on Its in vitro Transfection Efficiency HAO Jun-guo, SHA Xian-yi, JIANG Ye, FANG Xiao-ling* （Division of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai 200032, China ） Abstract: OBJECRIVE to research and develop a non-virus gene delivery system with high efficiency and safety; METHODS the third generation of polypropyleneimine(PPI) dendrimer modified with Pluronic P123 was synthesized by covalent linking, the size and ? potential of nanoparticles formed by plasmid condensing with P123-PPI/P123 were characterized, the ability of anti-dissociation to serum and sodium heparin was investigated with agarose gel electrophoresis, DNaseI protection assay was carried out to check whether the nanoparticles can protect plasmid DNA from nuclease digestion or not. the in vitro transfection efficiency was inspected with the reporter plasmid the enhanced green fluorescense plasmid (pEGFP-N2); RESULTS the nanoparticles formed by plasmid condensing with P123-PPI/P123 sized about 100 nanometers and its ? potential was around 15 millivolts ; CONCLUSION As a non-virus delivery vector, the th