一种载基因阳离子微泡的制备及与SonoVue的对比研究  

Preparation of a New Type of Cationic Microbubble Carrying Gene and Comparing with Sonovue

作　　者：杨未晓[1] 王翔[1] 杨丹[1] 谭开彬[1] 刘平[1] 高云华[1] 

机构地区：[1]解放军第三军医大学新桥医院超声诊断科,重庆市400037

出　　处：《中国超声医学杂志》2013年第4期371-374,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金青年科学基金项目(No.30901389)

摘　　要：目的制备一种表面带正电荷的阳离子微泡，与SonoVue对比研究其一般物理特性及对基因的携载能力，以期寻找一种更有效的基因载体。方法采用混悬-冻干-机械振荡法制备阳离子微泡，检测其大小、浓度和表面电位等物理特性，并与SonoVue相比较。以激光共聚焦显微镜观察阳离子微泡和SonoVue分别与质粒DNA的连接情况。将阳离子微泡和SonoVue分别与质粒DNA按不同体积比例分组孵育，以琼脂糖凝胶电泳观察共孵育后各组微泡对质粒DNA的携载能力。结果阳离子微泡呈规则球形，粒径较均匀，大小约2．7～6．8μm，浓度6．5×10^8个／ml，表面zeta电位（50．2±3．6）mV，SonoVue表面zeta电位为（-40．5±2．8）mV，二者差异显著。激光共聚焦显微镜下，较多发红色荧光的质粒DNA吸附在阳离子微泡的表面，呈“花环”状聚集，而与SonoVue共孵育的质粒在镜下呈杂乱分布。电泳结果显示，当阳离子微泡与质粒DNA体积比≥2：1时，质粒DNA完全滞留于加样孔中；反之，则随着二者体积比例的减小而逐渐逸出。而所有SonoVue与质粒DNA共孵育组，质粒均完全逸出加样孔。结论自制阳离子微泡的基因携载能力明显优于SonoVue，可作为基因载体研究提供一条新的思路。Objective To prepare a new type of cationic microbubble（CMB） and examine its ability for carrying gene comparing with SonoVue in order to find a more efficient gene vector. Methods The CMB was prepared by freeze-drying and mechanical shaking. Its morphology, size and concentration were observed and measured under the microscope. The zeta potential of CMB and SonoVue was measured by Malvern Zetasizer Nano. The CMB and SonoVue were incubated respectively with plasmid being stained by PI at room temperature. The morphology and con- nect effect of complex were oberserved by laser scanning confocal microscope. Furthermore, the complex incubated by defferent proportions was added in electrophoresis apparatus to examine the ability which CMB or SonoVue combined with plasmid. Results The mean size and concentration of CMB were 2.7-6.8 μm and 6.5 × 10^8/ml. The mean zeta potential of CMB and SonoVue was （50.2±3.6） mV and （-40.5±2.8） mV respectively. It showed that the wall of CMB was not smooth and a number of red plasmid surrounded it which looked like a wreath under confocal imaging. Instead, the wall of SonoVue was still smooth and the red plasmid displayed randomly under confocal imaging. It was also demonstrated that when the volume ratio of CMB to plasmid was greater than or equal to 2/1, the plasmid was combined by CMB and stayed in the well of etectrophoresis apparatus completely. But, when the volume ratio was less than 2/1 and decreased gradually, the plasmid had begun to run out of the well until all of the plasmid went out and appeared on the electrophoresis road. By contrast, there was no plasmid in the well of electrophoresis apparatus at the any volume ratio of SonoVue to plasmid. Conclusions The self-made CMB has higher ability of carrying gene than SonoVue and provides a new way on gene vector research.

关 键 词：微泡 基因 SONOVUE 

分 类 号：R943[医药卫生—药剂学]