载基因壳聚糖纳米粒的制备及其特性的研究  

Preparation and characteristics of chitosan nanoparticles as a gene vector

作　　者：赵惠萍[1] 马瑜[1] 边云飞[2] 高奋[2] 何军华[2] 肖传实[2] 

机构地区：[1]山西医科大学第二医院干细胞实验室,太原030001 [2]山西医科大学第二医院心内科,太原030001

出　　处：《中国药物与临床》2008年第10期774-776,共3页Chinese Remedies & Clinics

基　　金：山西省归国人员重点基金资助项目(2007-5)

摘　　要：目的制备壳聚糖纳米粒,并连接上质粒,研究壳聚糖纳米粒的特性及其对DNA的结合及保护能力。方法采用离子交联法制备壳聚糖纳米粒,并用喷金扫描电子显微镜检测,了解粒径的分布与形态;通过静电吸附作用连接上pGenesil-1质粒(报告基因);经琼脂糖凝胶电泳分析壳聚糖纳米载体与质粒DNA的结合能力,及不同pH值的壳聚糖纳米粒对质粒DNA的结合能力;并通过DnaseⅠ消化壳聚糖纳米-质粒结合物以观察壳聚糖纳米载体对质粒的保护作用。结果喷金扫描电镜检测证实壳聚糖纳米粒呈均匀分散的球形颗粒,平均直径为5nm;琼脂糖凝胶电泳的结果显示壳聚糖纳米粒能有效地结合载体pGenesil-1质粒;不同pH值的壳聚糖纳米粒对质粒的保护作用不同,当pH值<7时壳聚糖纳米载体能100%结合质粒;DnaseⅠ消化试验证实壳聚糖纳米载体对质粒DNA有保护作用。结论采用离子交联法制备出粒径较小、均匀的壳聚糖纳米粒,并且壳聚糖纳米粒能有效地连接上质粒并对其有保护作用。Objective To prepare the plasmid-binding chitosan nanoparticle （CS-NP） and explore its biologic properties, DNA affinity and protection. Methods Ion cross-linkage was used to prepare CS-NP, and the diametric distribution and morphology of CS-NP were tested by gold-labeling scanning electron microscopy （SEM）. The binding of pGenesil-1 （reporter gene） to CS-NP was achieved by static electric attraction. The affinity and protective effect of CS-NP for plasmid under different pH conditions was analyzed with agarose gel electrophoresis. Furthermore, DNA- protecting capacity of CS-NP complex was confirmed with Dnase I digestion. Results Under gold-labeling SEM, CS-NP were round, even-distributed nanoparticles measuring 5 nm in diameter. Agarose gel electrophoresis showed effective uptake of pGenesil-1 by CS-NP, while the protective action of CS-NP on plasmid varied with different pH conditions. At pH 〈7, nearly 100% of plasmids were CS-NP-bound. Protective action of CS-NP on plasmid was confirmed by Dnase Ⅰ . Conclusion CS-NP prepared by ion cross-linkage appeared as tiny, even-distributed particles that can effectively binds to plasmids to protect DNA.

关 键 词：壳聚糖 质粒 纳米粒 

分 类 号：R318.08[医药卫生—生物医学工程]