纳米谷氨酸壳聚糖制备及其体外质粒转染研究  

Preparation of Glutamic Acid-modified Chitosan Nanoparticles and Gene Transfection Research in vitro

作　　者：何琼琼[1] 肖旭贤[2] 冯德云[1] 

机构地区：[1]中南大学基础医学院病理教研室,长沙410078 [2]中南大学化学化工学院,长沙410083

出　　处：《科技导报》2008年第16期34-37,共4页Science & Technology Review

基　　金：国家自然科学基金项目(30671846);湖南省自然科学基金项目(07JJ4008)

摘　　要：采用表面修饰方法制备出谷氨酸修饰的壳聚糖纳米基因载体。对样品进行红外分析、粒度分析、zeta电位分析、生物相容性、凝胶阻滞分析、DNA保护性试验、体外细胞转染研究。结果显示所制得的谷氨酸修饰的壳聚糖纳米颗粒平均粒径为170nm,其zeta电位为+4.7mV。红外分析显示谷氨酸已通过酰胺键结合在壳聚糖上。MTT实验结果显示纳米颗粒与细胞有良好的生物相容性。凝胶阻滞分析和DNA保护试验结果表明纳米载体可与DNA通过电性结合作用而结合,并可以有效保护DNA,防止核酸酶对其的降解作用。而体外细胞转染的结果表明,谷氨酸修饰的纳米粒能介导pEGFP-N1质粒转染HepG2细胞并在细胞中表达绿色荧光蛋白。因此,谷氨酸修饰的壳聚糖纳米颗粒可作为一种新型非病毒基因载体介导核酸类生物大分子进入细胞内。Glutamic acid-modified chitosan nanoparticles （GMCN） as gene vectors were synthesized by surface-modifying technique and studied by using IR analysis, granularity analysis, zeta-potential measurement, biocompatibility, gel retardation assay, DNA protection assay and in vitro transfection. The results show that the mean diameter of the GMCN is 170 nm and the zeta potential is about ＋4.7 mV. IR analysis reveals that glutamic acid has been combined with chitosan nanoparticles by amide linkage. MTT assay shows that the nanoparticles have excellent biocompatibility with cells. The results of gel retardation analysis and DNA protection assay show that GMCN can completely combine with the DNA by electronic combination and protect DNA against serum degradation. The in vitro transfection of GMCN was effective in HepG2 cells and the expression of green fluorescent proteins was observed with fluorescent microscope. The GMCN may serve as a new-type and effective non-viral carrier for delivering nucleotides into cells.

关 键 词：谷氨酸 修饰 纳米 壳聚糖 基因载体 

分 类 号：Q782[生物学—分子生物学]