P123-PPIDNA纳米粒性质表征和体外转染效率研究.doc

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ζ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，ζ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 ζ 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 ζ potential was around 15 millivolts ; CONCLUSION As a non-virus delivery vector, the third generation of PPI dendrimer modified with Pluronic P123 possessed a high in vitro transfection efficiency and may be developed into a effective in vivo transfection vector. Key Words: gene therapy; Pluronic P123; polypropyleneimine dendrimer 基金项目：国家重大基础研究项目973项目（2007CB935800） 作者简介：郝俊国，男，复旦大学药物制剂专业在读博士研究生 通讯作者：方晓玲