高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (9): 1896.

• 研究论文 • 上一篇    下一篇

六氯环三磷腈交联寡聚乙烯亚胺的制备及基因载体应用

李非凡1,2, 田华雨1, 陈磊1,2, 夏加亮1,2, 陈学思1, 景遐斌1   

  1. 1. 中国科学院长春应用化学研究所, 生态环境高分子材料重点实验室, 长春 130022
    2. 中国科学院研究生院, 北京 1000497
  • 收稿日期:2009-11-24 出版日期:2010-09-10 发布日期:2010-09-10
  • 通讯作者: 陈学思, 男, 博士, 研究员, 博士生导师, 主要从事生物医用高分子和生物降解高分子研究. E-mail: xschen@ciac.jl.cn
  • 基金资助:

    国家自然科学基金(批准号: 20604028, 50873102, 50733003, A3项目50425309)和科技部项目(批准号: 2005DFA50290, 2007DFR50200)资助.

Synthesis and Characterization of the Hexachlorotriphosphaxene Cross-linked Oligo(ethyleneimine) as Gene Carrier

LI Fei-Fan1,2, TIAN Hua-Yu1, CHEN Lei1,2, XIA Jia-Liang1,2, CHEN Xue-Si1*, JING Xia-Bin1   

  1. 1. Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
    2. Graduate University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2009-11-24 Online:2010-09-10 Published:2010-09-10
  • Contact: CHEN Xue-Si. E-mail: xschen@ciac.jl.cn
  • Supported by:

    国家自然科学基金(批准号: 20604028, 50873102, 50733003, A3项目50425309)和科技部项目(批准号: 2005DFA50290, 2007DFR50200)资助.

摘要: 将六氯环三磷腈与分子量为600的超支化寡聚乙烯亚胺在干燥氯仿中反应, 合成了可降解的交联型聚合物. 利用1H NMR表征了聚合物的结构, 并用GPC测试了聚合物的分子量. 研究了其作为非病毒基因载体的性能, 聚合物载体与DNA形成的复合物颗粒粒径为150 nm, Zeta电位为30~40 mV, 凝胶阻滞电泳显示聚合物/DNA在质量比为0.4时能够将DNA完全阻滞. 体外转染实验结果表明, 载体对HeLa细胞的最佳转染效率为PEI-25K的3倍; 聚合物浓度为20 μg/mL时, 细胞存活率仍然大于80%, 材料的细胞毒性低, 生物相容性好, 具有良好的生物医学应用前景.

关键词: 六氯环三磷腈, 寡聚乙烯亚胺, 基因载体, 转染, 细胞毒性

Abstract: Hyperbranched oligo(ethyleneimine) with molecular weight of 600 was carried out reaction with hexachlorotriphosphazene in anhydrous chloroform to synthesize a degradable and cross-linked polymer as a non-viral gene carrier. The 1H NMR spectra confirmed the formation of the polymer and its average molecular weight was obtained by GPC with HAc/NaAc buffering solution of pH=4.4 as fluent phase. The polymer compressed DNA into stable complexes in diameter of about 150 nm and zeta potential of 30—40 mV measured by dynamic light scattering. Besides, when the mass ratio of polymer and DNA was 0.4, the plasmids were fully retarded in 1% agarose gel by gel retardation electrophoresis. The cytotoxicity and transfection efficiency were characterized by HeLa cells in vitro. It was indicated that the best transfection efficiency of polymer with lower cytotoxicity was three times higher than that of PEI-25K. Therefore, the synthesized polymer may have good potential applications as non-viral gene carriers.

Key words: Hexachlorotriphosphazene, Oligo(ethyleneimine), Gene carrier, Transfection, Cytotoxicity

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