寡肽阳离子脂质体作为基因载体的体外研究  

In vitro evaluation of cationic liposomes prepared with oligopeptide-based lipids for plasmid DNA delivery

作　　者：崔烨[1] 王璐[1] 孙琼[1] 张灿[1] 

机构地区：[1]中国药科大学新药研究中心,南京210009

出　　处：《中国药科大学学报》2013年第4期328-333,共6页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81072589;No.81273468)~~

摘　　要：采用阳离子寡肽脂质材料赖氨酰化谷氨酸双十四醇酯(T2GL)制备阳离子脂质体(CL),与增强型绿色荧光蛋白质粒(pEGFP)静电结合形成复合物(CL/pDNA),CL/pDNA粒径和表面电位分别为(132.3±1.0)nm和+(31.35±2.99)mV。考察CL/pDNA对肝素及脱氧核糖核酸酶Ⅰ(DNase I)的稳定性,结果表明,CL/pDNA可被适宜浓度的肝素竞争性解组装释放pDNA,其对DNase I的稳定性明显优于市售制剂Lipofectamine 2000。研究CL与pDNA的比例(N/P)及质粒用量对转染效率的影响,结果显示当N/P为3,质粒用量4μg时,CL/pDNA的转染效率最佳。采用CL/pDNA对3种不同的细胞进行转染,发现CL/pDNA在人源非小细胞肺癌A549细胞中转染效果最好,与Lipofectamine 2000相比,转染效率相当但细胞毒性更低,因此为进一步研究其体内行为提供了良好的基础。Cationic liposomes（CL） were prepared using ditetradecyl N-lysyl-L-glutamate（T2GL） as cationic lipid and soybean lectithin as helper lipid.Lipoplexes（CL/pDNA） were formed between the CL and plasmid EGFP due to electrostatic interaction.The particle size of CL/pDNA was assayed to be（132.3±1.0） nm while Zeta potential was ＋（31.35±2.99）mV.Both CL and CL/pDNA were unilamellar vesicles in terms of morphology.The release of pDNA from CL/pDNA occured when incubated with the appropriate concentration of heparin.When incubated with DNase I,there was no change to CL/pDNA but there existed degradation of pDNA,either naked or bound with Lipofectamine 2000.CL/pDNA was found to possess the highest transfection efficiency（TE） at N/P ratio of 3 and pDNA weight of 4 μg.TE of CL/pDNA was evaluated using 3 kinds of cell lines.In A549 cells,CL/pDNA yielded almost equal TE but much lower cell toxicity if compared with Lipofectamine 2000 reagent.Therefore,it suggested that CL/pDNA be a promising genetic vehicle worth of in vivo evaluation.

关 键 词：寡肽 阳离子脂质体 质粒 体外稳定性 转染效率 细胞毒性 

分 类 号：R944[医药卫生—药剂学]