超声辐照微泡声学造影剂增强脂质体介导肝细胞基因转染的体外实验研究  

Ultrasound-mediated Microbubble Destruction Enhanced Liposome Gene Transfection in Liver Cells:in Vitro Study

作　　者：苏琳[1] 王志刚[1] 李兴升[1] 任建丽[1] 汪朝霞[1] 李雪霖[1] 许川山[1] 张茂惠[1] 

机构地区：[1]重庆医科大学超声影像学研究所,重庆医科大学附属第二医院超声科,重庆市400010

出　　处：《中国超声医学杂志》2007年第10期721-723,共3页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金重点项目(No.30430230);重庆自然科学基金重点项目(CSTCNo.2005BA5024);国家863科技攻关项目[国科发财字No.(2006)501]

摘　　要：目的研究超声破坏微泡声学造影剂提高脂质体介导肝细胞生长因子质粒(pIRES-EGFP-HGF)在肝细胞中转染率的可行性。方法将培养的肝细胞分为4组:(1)单纯对照组,(2)脂质体转染组,(3)超声辐照脂质体转染组,(4)超声辐照微泡+脂质体转染组。第(4)组按照每孔加入造影剂剂量又分为1)10μl组,2)20μl组,3)30μl组,4)40μl组4亚组。超声辐照微泡+脂质体转染组在脂质体介导肝细胞生长因子转染肝细胞1h后,在24孔板中每孔加入微泡声学造影剂10,20,30或40μl,超声辐照60s。24h后用荧光显微镜观察绿色荧光蛋白表达情况、MTT法检测肝细胞增殖率和流式细胞仪检测基因转染率。结果超声频率1MHz,功率0.5W/cm2辐照60s,每孔加入造影剂30μl时肝细胞基因转染效率最高,与脂质体转染组比较有显著性。结论在一定条件下,超声辐照微泡声学造影剂可明显提高脂质体介导肝细胞基因转染效率,为基因治疗提供了新的思路。Objective To investigate whether ultrasound mediated mierobubble destruction could effectively enhance the efficiency of liposome delivery plasmid to L 02 cells. Methods The cultured L 02 cells were divided into four groups. The first group was the control group and the second group was given proper dose of liposome with plasmid. Liposome was added with plasmid to the third group and exposed to ultrasound. Liposome with plasmid and microbubbles was applied to the fourth group and exposed to ultrasound. As for the microbubble dosage, the fourth group was devided into 4 subgroups： 1- 10 μl, 2.20 μl, 3.30 μl and 4.40 μl groups. One hour after liposome delivered plRES EGFP EHGF to L 02 cells, the fouth group was added 10 μl, 20 μl, 30 μl or 40 μl microbubbles into each well and exposed to ultrasound. After 24 hours, the plRES-EGFP HGF expression in the L 02 cells was detected by fluorescence microscopy, MTT and flow cytometry. Results After exposed to the 1 MHz, 0. 5 W/cm^2 uhrasound for 60 s, 30 μl microbubble were added into each well in the 4^th group, significantly enhanced efficiency of liposome plasmid transfection was observed. Conclusions In certain condition, the liposome delivered plRES-EGFP HGF expression efficiency in L 02 cells was increased with the administration of uhrasound-mediated mierobubbles destruction.

关 键 词：超声 微泡 肝细胞 脂质体 基因转染 

分 类 号：R346[医药卫生—基础医学]