两种非病毒载体与质粒DNA的结合及解离过程的停流动力学研究  

Stopped-flow kinetic studies of the association and dissociation of plasmid DNA with two kinds of nonviral gene delivery systems

作　　者：刘瑞珍[1] 孙晓辉[1] 甘璐[1] 

机构地区：[1]华中科技大学生命科学与技术学院,湖北武汉430074

出　　处：《中国医院药学杂志》2008年第14期1143-1146,共4页Chinese Journal of Hospital Pharmacy

基　　金：国家863资助项目(编号:2004AA2Z3212)

摘　　要：目的:对由1,2-二油烯氧基-3-三甲氨基丙烷[1,2-dioleoyl-3-trimethy-lammonium-propane(chloride salt),DOTAP]、二油酰基磷脂酰乙醇胺(1,2-Dioley-l-s-Glycero-3-Phosphoeth anolamine,DOPE)形成的阳离子脂质体与质粒DNA及壳聚糖与质粒DNA的结合及解离动力学过程进行研究。方法:以钌联吡啶配合物[Ru(phen)2dppz]2+作为荧光探针采用荧光停流光谱技术研究两种载体同DNA的结合和解离动力学。结果:结合动力学过程中,随N/P(载体中伯氨基数目/pDNA中磷酸基数目)比的增加,荧光强度的变化幅度增大,在N/P比小于4时,结合速率常数k1>k2。在解离动力学过程中,随N/P比的增加,荧光强度的变化越来越小,在所选N/P比范围内,解离速率常数k1>k2。结论:DNA压缩是复合物形成过程中的限速步骤,而DNA的伸展及重排是复合物解离过程中的限速步骤。OBJECTIVE To investigate the association and dissociation kinetics of DNA with two kinds of nonviral gene delivery systems （cationic liposome formed by 1,2-dioleoyl-3-trimethy larnmoniurn propane （chloride salt） （DOTAP） and 1,2 Di oleyl-s-Glycerc-3-Phosphoeth anolamine （DOPE） and chitosan）. METHODS Stopped flow fluorescent spectrophotometry was used to investigate the association and dissociation kinetic of DNA with two kinds of nonviral vectors using [Ru（phen）2dppz]^2＋ as a probe. RESULTS With the increasing of N/P ratio, the fluorescence intensity and the rate constant decreased in the process of association; while the opposite results were obtained during the dissociation process. CONCLUSION The results indicat that DNA condensation is the rate-determining step during the complexation process, while DNA extending and rearrangement is the rate-determining step during the DNA dissociation process.

关 键 词：DOTAP/DOPE 壳聚糖 DNA 结合 解离 停流动力学 

分 类 号：R318.01[医药卫生—生物医学工程]