CAJ | 학술논문

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PEI-RAP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砄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。
목적：제비취을희아알（ PEI）-황기다당（ RAP）공취물，탐토기작위비병독기인재체적가행성。방법：채용담병정법、고전산갑-PEI법이급탄기이미서-PEI법제비PEI-RAP。이용홍외광보감정3충방법소득산물적결구표정。통과경지당응효전영이급립경화Zeta전위적측정고찰PEI-RAP여질립DNA적상호작용。 MTT법고찰PEI-RAP재체적세포독성。 PEI-RAP부재록색형광표체재체pEGFP전염MCF-7、Hela화SMMC-7721세포주관찰전염효능。결과：통과담병정법성공지장PEI접지재RAP상，단시령외2충방법미득도목표산물。경지당응효전영결과표명PEI-RAP가이통과정전작용부재질립 DNA。당PEI-RAP여pEGFP 질량비위12결1시，복합물립경위156．61 nm，전위위＋26．89 mV。 PEI-RAP재체무세포독성。체외전염실험표명，PEI-RAP가장pEGFP고효전염지MCF-7、Hela、SMMC-7721세포。결론：성공제비료PEI-RAP공취물，해공취물시일충구유잠재응용전경적신적비병독기인재체。
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.