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壳聚糖接枝PEI非病毒基因载体的均相合成及其基因转染
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摘要
基因治疗的首要问题是开发安全、高效的基因载体~([1])。壳聚糖基因载体生物安全性好,但转染效率有待提高~([2])。但壳聚糖在非水介质中的难溶性,使其改性修饰反应受到诸多限制。咪唑类离子液体1-丁基-3-甲基咪唑醋酸盐([BMIM]Ac)能够有效克服壳聚糖的溶解性问题,是实现壳聚糖非水介质中均相、高效改性修饰的有效介质之一~([2,3])。本研究以[BMIM]Ac为反应介质,对壳聚糖进行聚乙烯亚胺(PEI)高效、可控接枝,得到壳聚糖接枝聚乙烯亚胺(CS-g-PEI-x),采用核磁共振波谱和红外光谱技术对化合物的结构进行了确定;在人喉癌上皮细胞Hep-2细胞株中对合成接枝聚合物载体的基因转染等生物学性能进行了评价,绿色荧光蛋白基因转染结果表明,聚乙烯亚胺接枝壳聚糖的转染效率较壳聚糖和PEI均有很大提高。本研究结果表明采用离子液体[BMIM]Ac为反应介质得到的壳聚糖接枝聚乙烯亚胺有望成为基因治疗的优良载体。
The main problem of gene therapy is the lack of safe and efficient gene vector.Chitosan is safe,but the ransfection efficiency should be improved.The insolubility of chitosan in non-aqueous medium limits the modification.Ionic liquid l-butyl-3-methyl imidazole acetate([BMIM]Ac) can dissolve chitosan and make the modification reaction conducted homogeneously and efficiently.Herein,chitosan grafted PEI was conducted in[BMIM]Ac by an efficient and controllable reaction.HNMR and FTIR was used to determine the structure of the grafted copolymers.Gene transfection efficiency was evaluated in human Hep-2 cell lines.The results suggested that the synthesized copolymers in[BMIM]Ac were prospective in gene therapy.
引文
[1]Cassiday,L.Nature 2014,509:651.
    [2]Chen,H.;Cui,S.;Zhao Y.;et al.Plos One 2015,10(4):e0121817.
    [3]张树彪,陈会英,崔韶晖,等.中国发明专利,201410043960.6,2014.

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