磁性氧化铁纳米颗粒与脂质体介导EGFP基因转染内皮祖细胞的比较研究

目的评价精氨酸包裹的超顺磁性氧化铁纳米颗粒（nano-Fe2O3-arginine）及脂质体作为基因载体在针对外周血来源的内皮祖细胞转染中的可行性,并对两者的转染效率进行比较。方法将nano-Fe2O3-arginine及LipofectamineTM2000作为基因载体连接绿色荧光蛋白pcDNA3.1（＋）/EGFP质粒在体外转染兔外周血来源的内皮祖细胞,荧光显微镜观察转染结果并计算转染效率;转染后48h采用MTT比色法测定这两种载体对内皮祖细胞增殖和功能的影响以了解其细胞毒性。结果LipofectamineTM2000作为基因载体将报告基因转染至兔外周血内皮祖细胞内并成功表达,用荧光显微镜可观察到发绿色荧光的细胞,转染效率达38.3%;而nano-Fe2O3-arginine转染组在荧光显微镜仅能观察到少量绿色荧光细胞,转染效率不到5%;两种载体对内皮祖细胞的增殖活性未见明显影响。结论脂质体作为载体转染内皮祖细胞的效率明显高于精氨酸包裹的超顺磁性氧化铁纳米颗粒,脂质体可较为有效安全地转染基因至内皮祖细胞,从而为内皮祖细胞联合基因治疗疾病奠定了基础。

A compared study of SPIO-mediated and liposome-mediated EGFP gene transfer to endothelial progenitor cells

Objective This study was designed to evaluate the feasibility of using arginine coated superparamagnetic iron oxide nanoparticles (nano-Fe₂O₃-arginine) and liposome as gene vectors in the gene transfection of endothelial progenitor cells (EPCs) derived from peripheral blood of rabbits, and compare their transfection efficiency. Methods nano-Fe₂O₃-arginine and Lipofectamine^TM 2000 combined with plasmid pcDNA3.1(+)/EGFP as the reporter gene were transfected into EPCs in vitro, the EGFP expression in EPCs was observed by fluorescence microscopy and the transfection efficiency was calculated. 48 h after transfection, MTT was used to evaluate the effect of the two gene carriers on the proliferation and function of cells, as well as acknowledge of their toxicity. Results Lipofectamine^TM 2000 can transfect reporter gene into EPCs and EGFP was successfully expressed, and the transfection efficiency was about 38.3%; while the transfection efficiency of nano-Fe₂O₃-arginine was less than 5%. The two gene carriers had no significantly toxicity on EPCs. Conclusion The efficiency of liposome to transfer gene into EPCs in vitro was significantly higher than that of nano-Fe₂O₃-arginine in this study. Liposome can transfer gene to EPCs effectively and safely, which provides theoretical support for further therapeutic alliance of gene and EPCs.