摘要
目的制备聚阳离子纳米粒非病毒基因载体:葡聚糖-寡胺化合物。方法以葡聚糖、过碘酸钠、精胺等为反应物,经多步化学合成制备了该载体;并以红外光谱测定仪、激光纳米粒度测定仪、透射电子显微镜对其进行了理化表征;以绿色荧光蛋白基因为报告基因考察所制备载体的基因转染效能。结果葡聚糖与寡胺间以共价键连接成稳定的纳米粒基因载体,载体平均粒径为9.0 nm,Zeta电位为+31.1 mV、形状为类圆形;载体具有较高的基因转染率。结论实验结果表明,文中方法可以合成出优良的聚阳离子纳米粒非病毒基因载体。
OBJECTIVE To prepare the dextran-sperminee polycation as the nonviral gene carrier.METHODS The carrier was prepared by the chemical synthesis, using the dextran, sodium periodate, spermine and sodium borohydride as the reactants. The main physical properties of the synthesized gene carrier were characterized by the methods of FTIR spectrophotometer, transmission electron microscope and laser nanoparticle detecter. The transfection with the dextran-spenninee polycation was performed using the pCMV-GFP as the reporter gene, the Transffectine 2000 liposome as the positive controller, the SMMC-7721 as transfected cells. The transfecting result was analyzed by fluorescence microscope instrument.The yield of transfection （ %transfection）was calculated by counting the fluorescent cells in a certain area of a particular well, and dividing the number of fluorescent cells by the number of total cells in the same field.RESULTS The dextran and spermine were linked by the covalent bond.The size of the carrier was 9.0 nm, the Zeta potential was ＋ 31.1 mV. The carrier had a higher transfection efficency to SMMC-7721 cells.The yield of transfection was 31.1%. CONCLUSION The chemical synthesis method mentioned above can be used to prepare the optimum dextran-spermince polycation nanoparticle as nonviral gene carrier.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2006年第13期1005-1009,共5页
Chinese Pharmaceutical Journal
基金
国家自然科学基金资助项目(30270400)
