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寡肽阳离子脂质体作为基因载体的体外研究

崔烨, 王璐, 孙琼, 张灿

崔烨, 王璐, 孙琼, 张灿. 寡肽阳离子脂质体作为基因载体的体外研究[J]. 中国药科大学学报, 2013, 44(4): 328-333. DOI: 10.11665/j.issn.1000-5048.20130408
引用本文: 崔烨, 王璐, 孙琼, 张灿. 寡肽阳离子脂质体作为基因载体的体外研究[J]. 中国药科大学学报, 2013, 44(4): 328-333. DOI: 10.11665/j.issn.1000-5048.20130408
CUI Ye, WANG Lu, SUN Qiong, ZHANG Can. In vitro evaluation of cationic liposomes prepared with oligopeptide-based lipids for plasmid DNA delivery[J]. Journal of China Pharmaceutical University, 2013, 44(4): 328-333. DOI: 10.11665/j.issn.1000-5048.20130408
Citation: CUI Ye, WANG Lu, SUN Qiong, ZHANG Can. In vitro evaluation of cationic liposomes prepared with oligopeptide-based lipids for plasmid DNA delivery[J]. Journal of China Pharmaceutical University, 2013, 44(4): 328-333. DOI: 10.11665/j.issn.1000-5048.20130408

寡肽阳离子脂质体作为基因载体的体外研究

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

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

  • 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4 μg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。
    Abstract: 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.
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出版历程
  • 刊出日期:  2013-08-24

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