壳聚糖纳米基因载体的制备及特性的研究  

Preparation and characteristics of chitosan-DNA nanoparticles as gene carriers

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

机构地区：[1]山西医科大学第二临床医学院心内科,太原030001

出　　处：《山西医科大学学报》2009年第1期10-12,共3页Journal of Shanxi Medical University

基　　金：山西省归国人员重点基金资助项目(2007-5)

摘　　要：目的制备适当粒径的壳聚糖纳米粒,并连接上质粒,研究壳聚糖纳米粒对质粒DNA的结合能力及保护作用。方法采用离子交联法制备壳聚糖纳米粒,通过喷金电镜观察其大小、形态及分布;经琼脂糖凝胶电泳分析纳米载体与DNA的结合能力及对DNA的保护作用;通过紫外分光光度计检测其包埋率。结果喷金电镜证实壳聚糖纳米粒分布均匀,呈近似球形,平均粒径约5nm。琼脂糖凝胶电泳结果显示壳聚糖纳米粒能有效地结合质粒,并保护DNA免受DnaseⅠ的降解,紫外分光光度计检测按不同比例结合质粒的壳聚糖纳米粒(纳米粒∶质粒)的包埋率分别为98.7%(10∶10),98.1%(10∶20),96.7%(10∶30),87.1%(10∶40),74.6%(10∶50)。结论成功制备了适当粒径且分布均匀的壳聚糖纳米粒。壳聚糖纳米粒能有效地结合质粒,并保护其免受DnaseⅠ的降解。Objective To explore the loading capability of chitosan nanopartieles and its protection ability to plasmid. Methods The chitosan nanoparticles were prepared with ionic cross-linking method. Its size, morphology and distribution were observed by scanning electron microscope. The binding ability and protective effect of pDNA were evaluated by agarose gel electrophoresis analysis, and the capsulating rate was determined by fluorospectrophotometry. Results Chitosan nanopartieles was found to be evenly distributed as spherical particles by scanning electron microscope with the mean diameter of about 5 nm. Agarose gel electrophoresis showed that chitosan nanoparticles were effectively combined with pDNA and protected it from nuclease degradation. The encapsulation efficiency of chitosan nanoparticles-DNA mixed by different ratios of nanoparticles： plasmid were 98.7% （ 10： 10） ,98.1% （ 10： 20） ,96.7% （ 10： 30）, 87.1% （ 10：40 ） ,74.6% （ 10：50 ） by fluorescent microscope, respectively. Conclusion Homogeneous in diameter ehitosan nanoparticles can be prepared, and the chitosan nanoparticle could load with plasmid effectively and protect pDNA from nuclease degradation.

关 键 词：壳聚糖 纳米粒 ACE-shRNAp Genesil-1质粒 包埋率 DnaseⅠ 

分 类 号：R318.08[医药卫生—生物医学工程]