bcl-xL纳米微粒的制备及其对大鼠心肌细胞的高效转染

Preparation of nanoparticles containing bcl-xL gene and its high efficient transfection in rat cardiac myocytes in vitro

目的：制备包载含人类抗凋亡基因bcl—xL的纳米微粒，评价其转染大鼠心肌细胞的能力．方法：用超声双乳化蒸发法制备包载bcl—xL的纳米微粒，扫描电镜观察纳米的形态和大小，DNase I消化酶试验检测其抗核酸酶能力；转染体外培养的大鼠心肌细胞后RT—PCR和Western—blot法检测基因表达．结果：制备的纳米微粒直径约200～260nm，包封完好，性能稳定并对基因有缓释作用；转染1wk后bcl-xL表达高于对照组（P〈0．05）．结论：成功制备包载真核表达质粒的纳米微粒，并能够高效转染大鼠心肌细胞．纳米微粒是心肌细胞转染较理想的基因载体之一．

AIM： To prepare nanoparticles containing bcl-xL gene and to investigate the feasibility of delivering the gene into rat cardiac myocytes in vitro. METHODS： Nanoparticles were prepared by the double emulsion-solvent evaporation technique, Its morphology and size were observed under scanning electron microscope and DNA protection was observed by DNase I digestion. After bcl-xL gene was transferred into cardiac myocytes by LipofectAMINE 2000 and nanosphere respectively, bcl-xL expression was detected by Western-blot and RT-PCR. RESULTS： Nanoparticles, with diameter ranging 200-260 nm, were well packaged, stable and capable of slowly releasing gene. bcl-xL expression in cardiac myocytes increased l week ＂after the transfection compared with that in the controls （P 〈 0. 05 ）, CONCLUSION： The nanoparticles containing bcl-xL gene have been successfully prepared and the exogenous bcl-xL can efficiently express in cardiac myocytes. Nanoparticle is an optimal gene vector for cardiac myocytes.