组氨酸修饰聚酰胺-胺型树状高分子提高血清中基因转染效率的研究  

Histidine Modified PAMAM as Gene Vector for Enhancing Gene Transfection Efficiency in Serum

作　　者：温玉婷[1] 潘仕荣[1] 郭振寰[1] 王持[2] 曾昕[2] 吴红梅[2] 冯敏[2] 

机构地区：[1]中山大学附属第一医院,心血管研究室,广州510080 [2]中山大学药学院,广州510080

出　　处：《中国生物医学工程学报》2010年第1期129-136,共8页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30870618)

摘　　要：组氨酸的咪唑环由于其独特的质子化机理而受到了广泛的关注。研究表明,组氨酸修饰的基因载体可对抗血清对基因转染的影响,提高载体在血清中的基因转染效率。本研究利用氨解反应将组氨酸接枝到聚酰胺-胺树枝状高分子(PAMAM)的表面,制备了一种新型的PAMAM衍生物—组氨酸修饰的聚酰胺-胺树枝状高分子(His-PAMAMG4)。利用1HNMR对His-PAMAM G4进行表征,证明一个PAMAM G4分子上接枝37个组氨酸分子。对His-PAMAM G4/DNA复合物进行结合DNA能力、粒径、表面电位、粒子形态等理化性能的表征,证明该新型聚合物对DNA分子具有良好的压缩能力。细胞活性检测结果表明,His-PAMAM G4对Bel 7402和Hela细胞的毒性均显著低于未经修饰的PAMAM G4。His-PAMAM G4在血清中的转染效率与PAMAM G4相比大幅度提高,也显著高于阳离子聚合物PEI 25k和市售商品阳离子脂质体Lipofectamine。因此,His-PAMAM G4有望成为一种高效、安全、可在体内应用的非病毒基因载体。The unique protonation mechanism of imidazole in histidine molecules attracts wide research attention in recent years. It has been reported by literatures that the impact of serum to the transfection efficiency could be reduced by modifying gene vectors with histidine to obtain enhanced transfeetion efficiency. In this work, histidine was conjugated to the surface of PAMAM G4 using aminolysis reaction to construct a PAMAM derivative （His-PAMAM G4） as a new gene vector. The chemistry of His-PAMAM G4 was characterized using 1H NMR, which showed that one PAMAM was linked with 37 histidine. The DNA binding ability, particle size, zeta potential, and the morphology of the His-PAMAM G4/DNA complexe were investigated. Experimental results revealed that the His-PAMAM G4 could sufficiently condense DNA. MTT assay indicated that the cytotoxicity of the His-PAMAM G4 was obviously lower than PAMAM G4 control to Bel 7402 ceils and Hela cells. The transfection efficiency of the His-PAMAM G4 detected by GFP flow cytometry was significantly enhanced in reference with that of PAMAM G4 control in serum. Therefore, the His-PAMAM G4 holds a potential of nonviral gene vectors in gene therapy in vivo.

关 键 词：聚酰胺-胺树枝状高分子 组氨酸 血清 基因转染 基因载体 

分 类 号：R318.08[医药卫生—生物医学工程]