壳聚糖脂质体复合载体核酸递送研究

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (8): 894-898.

壳聚糖脂质体复合载体核酸递送研究

王冰¹,崔韶晖¹,杨宝灵¹,赵轶男¹,赵不凋¹,周集体²,张树彪¹

1. 大连民族学院
2. 大连理工大学

收稿日期:2012-01-04 修回日期:2012-05-11 出版日期:2012-08-05 发布日期:2012-07-17
通讯作者: 张树彪 E-mail:zsb@dlnu.edu.cn
基金资助:
氨基酸型蔗糖酯类脂的合成及其运载siRNA的研究;复合型非病毒基因载体的构建及其运载机制研究

Chitosan and cationic liposome complex vector for gene delivery

Received:2012-01-04 Revised:2012-05-11 Online:2012-08-05 Published:2012-07-17

摘要/Abstract

摘要： 目的构建了一种由脂质体Lipofectamine2000、低分子量壳聚糖、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。

关键词: 壳聚糖, 脂质体, 复合载体, 基因递送体系

Abstract: Objective A new type of stable ternary complex was formed by mixing Lipofectamine2000 with chitosan/pDNA polyplex for delivery of plasmid DNA. Methods Morphologies of liposome/chitosan/pDNA were 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 microscope. Cell toxicity was evaluated by the MTT assay. Results Complex vector could combine pDNA and retard it completely. The liposome/polymer/pDNA complexes were incompacted 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 nearly no expression. Chitosan reduced cell toxicity of liposome. Conclusion New ternary complex has very higher transfection potential in gene delivery.

Key words: chitosan, liposome, complex vector, gene delivery system

王冰崔韶晖杨宝灵赵轶男赵不凋周集体张树彪. 壳聚糖脂质体复合载体核酸递送研究[J]. 基础医学与临床, 2012, 32(8): 894-898.

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