聚乙烯亚胺-黄芪多糖共聚物的制备及其基因传递性能观察  

Preparation of polyethylenimine-radix astragali polysaccharide copolymer and its perfermace as a gene delivery vector

作　　者：张丹丹[1] 任雪玲[2] 何阿妹 张红[2] 周宇雪[2] 张振中[2] 

机构地区：[1]河南省人民医院药学部,郑州450003 [2]郑州大学药学院,郑州450001

出　　处：《郑州大学学报（医学版）》2014年第4期457-460,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金81273451;国家级大学生创新创业训练计划项目201310459047

摘　　要：目的:制备聚乙烯亚胺(PEI)-黄芪多糖(RAP)共聚物,探讨其作为非病毒基因载体的可行性。方法:采用氮丙啶法、高碘酸钾-PEI法以及羰基二咪唑-PEI法制备PEI-RAP。利用红外光谱鉴定3种方法所得产物的结构表征。通过琼脂糖凝胶电泳以及粒径和Zeta电位的测定考察PEI-RAP与质粒DNA的相互作用。MTT法考察PEIRAP载体的细胞毒性。PEI-RAP负载绿色荧光表达载体pEGFP转染MCF-7、Hela和SMMC-7721细胞株观察转染效能。结果:通过氮丙啶法成功地将PEI接枝在RAP上,但是另外2种方法未得到目标产物。琼脂糖凝胶电泳结果表明PEI-RAP可以通过静电作用负载质粒DNA。当PEI-RAP与pEGFP质量比为12∶1时,复合物粒径为156.61 nm,电位为+26.89 mV。PEI-RAP载体无细胞毒性。体外转染实验表明,PEI-RAP可将pEGFP高效转染至MCF-7、Hela、SMMC-7721细胞。结论:成功制备了PEI-RAP共聚物,该共聚物是一种具有潜在应用前景的新的非病毒基因载体。To prepare polyethylenimine-radix astragali polysaccharide （PEI-RAP） copolymer and investigate the efficiency of PEI-RAP to transfect tumor cells in vitro .Met hods： PEI-RAP copolymers were prepared with three different methods, polymerization of aziridine, KIO4-PEI reactions and CDI-PEI reactions.The products were characterized using FT-IR.The interaction of PEI-RAP with DNA was investigated using particle size analysis , Zeta-potential measurements and gel electrophoresis assay .MTT method was used to detect the cytotoxicity of PEI-RAP.PEI-RAP was used to deliver pEGFP in three tumor cells, MCF-7, Hela and SMMC-7721 cell lines.Results：PEI-RAP was prepared by polymerization of aziridine successfully , while the other two methods were failed .Agarose gel retardation assay suggested that PEI-RAP could bind and condense plasmid DNA efficiently .When the mass ratio of PEI-RAP to the pEGFP was 12：1,the size and the Zeta potential of the complexes were 156.61 nm and ＋26.89 mV.The vector PEI-RAP showed no cytotoxicity .The reporter protein assay showed that pEGFP delivered by PEI-RAP could express green fluorescent protein efficiently in three tumor cells in vitro . Conclusion：The prepared PEI-RAP copolymer could be a promising non-viral gene delivery vector.

关 键 词：黄芪 多糖 共聚物 基因载体 聚乙烯亚胺 

分 类 号：Q782[生物学—分子生物学]