α-CD-PAMAM抗血清阳离子聚合物的构建及其作为基因载体的性能评价

发布时间：2019-03-04 19:37

【摘要】：本文合成了一种α-环糊精(α-cyclodextrin,α-CD)与聚酰胺-胺(polyamidoamine,PAMAM)的接枝聚合物(Cy D-G1)。~1H核磁测试结果表明,每个环糊精分子上平均接枝了6.4个PAMAM-G1分子。凝胶电泳结果显示,Cy D-G1可以有效结合DNA,并保护DNA免于核酸酶DNase I的降解。当载体与DNA复合物的N/P比为40时,可以压缩DNA形成平均粒径为120 nm左右的粒子,复合物表面的zeta电位约为+21 m V。该复合物可以在血清存在的条件下保持粒子的完整性并在360 min内稳定性良好。与对照品PEI-25K载体相比,Cy D-G1在高浓度时仍表现出较低的细胞毒性。将Cy D-G1与市售Lipofectamine 2000和PEI-25K对比转染发现,Cy D-G1/DNA复合物在多种细胞系中具有较高的转染率,而且转染水平不受血清影响。通过激光共聚焦观察并结合流式细胞分析表明,该阳离子聚合物介导DNA可以在4 h内有效进入细胞核内。上述结果证明,该阳离子聚合物作为一种非病毒型基因传递系统具有优良的性能以及体内给药应用的潜在可行性。
[Abstract]:A graft polymer (Cy D-G1) of 伪-cyclodextrin (伪-cyclodextrin, 伪-CD) and polyamide-amine (polyamidoamine,PAMAM) was synthesized in this paper. An average of 6.4 PAMAM-G1 molecules were grafted on each cyclodextrin molecule. The results of gel electrophoresis showed that Cy D-G1 could effectively bind to DNA, and protect DNA from degradation of DNase I. When the N / P ratio of the carrier / DNA complex is 40, the DNA can be compressed to form particles with an average particle size of about 120 nm, and the zeta potential on the surface of the composite is about 21 MV. The complex can maintain the integrity of the particles in the presence of serum and has a good stability in the range of 360 min. Compared with PEI-25K vector, Cy D-G1 showed lower cytotoxicity at high concentration. Comparing the transfection of Cy D-G1 with that of Lipofectamine 2000 and PEI-25K, it was found that Cy D-G1/DNA complex had a high transfection rate in many cell lines, and the transfection level was not affected by serum. Laser confocal observation combined with flow cytometry showed that the cationic polymer-mediated DNA could effectively enter the nucleus within 4 hours. These results show that the cationic polymer as a non-viral gene transfer system has excellent performance and potential feasibility of drug delivery in vivo.
【作者单位】：南方医科大学南方医院;广东药科大学;中山大学附属第一医院;
【基金】：国家自然科学基金资助项目(81373339,81173013)
【分类号】：R914.5

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