载ACEshRNA表达质粒壳聚糖纳米转染培养的大鼠血管内皮细胞实验性研究  

The experiment of the chitosan nanoparticles containing angiotensin converting enzyme short-hairpin RNA expression plasmid transfection on cultured rat vascular endothelial cells

作　　者：高奋[1] 朱国斌[1] 赵惠萍[1] 马喻[1] 边云飞[1] 肖传实[1] 

机构地区：[1]山西医科大学第二医院,030001

出　　处：《实用医技杂志》2009年第5期335-336,共2页Journal of Practical Medical Techniques

基　　金：山西省归国留学基金重点项目(2007005)

摘　　要：目的研究载血管紧张素转化酶短发卡RNA(ACEshRNA)表达质粒壳聚糖纳米颗粒对体外培养大鼠血管内皮细胞的转染效率。方法采用离子交联法制备壳聚糖纳米粒,喷金扫描电子显微镜鉴定;通过静电吸附复合ACEshRNA表达质粒,采用琼脂糖凝胶电泳分析载体与DNA结合能力;用体外培养大鼠血管内皮细胞转染实验,评价体外壳聚糖颗粒的转染能力。结果体外对培养的大鼠血管内皮细胞的转染实验显示,绿色荧光蛋白的表达,说明该壳聚糖有一定的转染效率,且在壳聚糖与质粒体积比为1∶3时转染率最高。结论本研究制备的载ACEshRNA表达质粒壳聚糖纳米载体,能在体外培养的大鼠血管内皮细胞实现有效转染,并且为进一步转染条件的优化奠定实验基础。Objective To study the transfeetion efficiency of expression plasmid.eontaining angiotensin converting enzyme short-hairpin RNA （ACEshRNA） chitosan nanoparticles on cultured rat vascular endothelial cells. Methods To prepare chitosan nanoparticles with ionic crosslinking method and identify with sprayed gold scanning electron microscope; By electrostatic adsorption to bind ACEshlLNA composite expression plasmid, then using agarose gel eleetrophoresis analysis of vector and DNA binding capacity; using cultured rat vascular endothelial cell transfeetion experiments in vitro to evaluate chitosan particles transfeetion capacity. Results The vitro of cultured rat vascular endothelial cell transfeetion experiments showed that the expression of green fluorescent protein, indicating that the Chitosan has a certain degree of transfeetion efficiency, and in the ehitosan with plasmid volume ratio is 1：3 at the highest rate of transfection. Conclusion This study prepared ehitosan nano-cartier containing ACEshRNA expression plasmid and in cultured rat vascular endothelial cells to achieve effective transfection. These transfeetion conditions built the foundation for the further experiment.

关 键 词：肽基二肽酶A 质粒 壳聚糖 内皮细胞 

分 类 号：R318.08[医药卫生—生物医学工程]