壳聚糖脂质体复合载体的核酸递送  

Chitosan and cationic liposome complex vector for gene delivery

作　　者：王冰[1,2] 崔韶晖[2] 杨宝灵[2] 赵轶男[2] 赵不凋[2] 周集体[1] 张树彪[2] 

机构地区：[1]大连理工大学环境与生命学院,辽宁大连116024 [2]大连民族学院生物化学工程国家民委-教育部重点实验室,辽宁大连116600

出　　处：《基础医学与临床》2012年第8期894-898,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(20876027;21046008);新世纪优秀人才支持计划(NCET-08-0654);中央高校基础科研基金(DC10020103)

摘　　要：目的构建一种由脂质体Lipofectamine2000、低分子质量壳聚糖、pDNA组成的三元新型复合载体用于核酸递送能力研究。方法复合物形态采用原子力显微镜轻敲模式下表征、载体与核酸结合能力采用凝胶延滞法表征,Hep-2细胞报告基因表达利用倒置荧光显微镜检测。细胞毒性研究采用3-甲基-2-噻唑硫酮(MTT)法。结果复合载体与pDNA结合能力强,可完全延滞pDNA。脂质体/壳聚糖/pDNA复合载体形态呈现出未完全压缩的球形,短棒状和不规则的聚集块。新型载体转染Hep-2细胞提高了绿色荧光蛋白报告基因的表达效率。与脂质体对照载体比较,基因转染效率提高了2～4倍,对照壳聚糖载体无明显转染效果。细胞毒性表明壳聚糖降低了脂质体的细胞毒性。结论基于脂质体的壳聚糖新型复合载体具有核酸递送潜力。Objective To construct a new type of stable ternary complex by mixing Lipofectamine2000 with ehitosan/pDNA polyplex for delivery of plasmid DNA. Methods Morphology of liposome/chitosan/pDNA was characterized by atomic force microscopy （AFM） in tapping model. Vectors could bind pDNA sufficiently, which can be measured by gel retarding. GFP gene expression in Hep-2 cells in vitro was imaged by inverted fluorescence micro- scope. Cell toxicity was evaluated by MTT assay. Results Complex vector did combine pDNA and retard it completely. The liposome/polymer/pDNA complexes were ineompaeted spheroids, short rod and irregular lump of larger aggregates in structure. The transfection efficiency of the lipopolyplexes showed higher GFP gene expression than Lipofectamine2000/pDNA and CTS/pDNA controls. It was 2-to 4-fold than Liposome/pDNA control, while CTS/pDNA had few expression. Chitosan reduced cell toxicity of liposome. Conclusions New ternary complex has very higher transfection potential in gene delivery.

关 键 词：壳聚糖 脂质体 复合载体 基因递送体系 

分 类 号：R943[医药卫生—药剂学]