亚微米级超声微泡造影剂的制备及体外基因转染效率的实验研究

目的 探索制备亚微米级超声微泡造影剂的方法 ,以GFP作为目的 基因验证其作为一种新型基因载体的可行性.方法 以高剪切分散法制备超声微泡造影剂,透射电镜及激光粒度分析仪检测其形态及粒径;将超声微泡造影剂与不同剂量的绿色荧光蛋白质粒PShuttle-IRES-hrGFP-1结合后转染HepG2细胞,利用荧光显微镜观察并检测其基因转染效率.结果 自制超声微泡造影剂为均匀分散的圆泡,粒径分布在282.2～415.7 nm之间,平均值为(335±5)nm,达到亚微米级;该微泡能将GFP基因成功转运到HepG2细胞内并高效表达,转染效率达32.61%±3.42%.结论 自制亚微米级超声微泡造影剂粒径小、分散均匀,并能成功转运外源DNA进入细胞内,可作为一种新型基因载体.

Experimental Study on Preparation and Gene Transfer of Submicron Ultrasound Microbubbles Contrast Agent

Objective To search for a preparing method of submicron ultrasound microbubbles contrast agent and its availability as a new gene carrier was evaluated.Methods High shear dispersing system was usedto prepare submicron ultrasound microbubbles contrast agent.The form and diameter were examined by transmission electron microscopy and laser particle size analyzer.The mixtures of microbubbles contrast agent and PShuttle-IRES-hrGFP-1 plasmid were transfected to HepG2 cells,then fluorescence microscope was used to evaluate the efficiency of objective gene transfection.Results Homemade submicron contrast agent was averagely dispersed as microbubbles with average diameter of（335±5）nm.Homemade microbubbles contrast agent could deliver green fluorescent protein into HepG2 and be expressed.Its transfected efficiency reached 32.61%±3.42%.Conclusions Submicron microbubbles contrast agent was averagely dispersed with submicron diameter.It could be used as a new gene carrier because it could transfect exogenous DNA into HepG2 cells.