阳离子超支化缩水甘油醚衍生物对提高腺病毒载体基因表达率的体外研究

目的 制备阳离子超支化缩水甘油醚衍生物HPG-AGE-cys (HPAC),将其包裹在腺病毒(rAd)表面形成复合物,探究其提高腺病毒基因表达效率的能力.方法 采用阴离子开环聚合反应制备超支化缩水甘油醚,并进一步阳离子衍生化得到HPAC,其结构经核磁确证;通过静电复合作用制备材料-腺病毒复合物,并测定其粒径和Zeta电位,用透射电镜观察其表面形态;考察了复合物被A549细胞、CHO细胞和MDCK细胞摄取的能力和细胞转导的能力.结果 成功合成了阳离子超支化缩水甘油醚衍生物HPAC,制备了HPAC-rAd复合物.其平均粒径为128.8 ±3.8 nm,Zeta电位为29.8 ±5.2 mV;与裸病毒rAd比较,复合物被细胞摄取的能力和细胞转导的能力均有显著提高.结论 阳离子超支化缩水甘油醚衍生物材料合成方法简单易行,制备的HPAC-rAd复合物能够显著提高病毒的基因效率,在腺病毒的基因传递方面具有广阔的应用前景.

Enhanced in vitro gene expression efficacy of the adenovirus mediated by cationic hyperbranched polyglycerol derivatives

OBJECTIVE To synthesize cationic hyperbranched polyglycerol derivatives HPG-AGE-cys (HPAC).To prepare HPAC-rAd complexes and evaluate the cellular uptake and transduction effects.METHODS The cationic hyperbranched polyglycerol derivatives was synthesized using opening multibranching polymerization and the structure was confirmed by 1HNMR.The HPAC-rAd complex was prepared with electrostatic interaction.The size and Zeta potential were measured.The efficiency of cellular uptake and transduction was evaluated on A549 cells,CHO cells and MDCK cells in vitro.RESULTS The cationic hyerbranched polyglycerol derivatives were successfully synthesized and HPAC-rAd complexes was prepared.The size of complex was 128.8 ± 3.8 nm with the Zeta potential of 29.8 ± 5.2 mV.Compared with the naked rAd,the cellular uptake and transduction of complexes both were much higher.CONCLUSION The synthesis of cationic hyperbranched polyglycerol derivatives is simple and feasible.The HPAC-rAd complexes could enhance the gene delivery of rAd,remarkably.It is a potential system for gene delivery of rAd.