声诺维联合超声辐照增强绿色荧光蛋白质粒转染血管内皮细胞的实验研究  

Ultrasound Exposure Mediated Transfection of pEGFP to HUVEC:the Enhancement Role of an Ultrasound Contrast Agent (SonoVue)

作　　者：汤庆[1] 徐辉雄[1] 吕明德[2] 岳殿超[3] 李宝金[2] 向邦德[2] 

机构地区：[1]中山大学附属第一医院超声波科,广州市510080 [2]中山大学附属第一医院肝胆外科,广州市510080 [3]中山大学附属第一医院核医学科,广州市510080

出　　处：《中国超声医学杂志》2005年第5期328-331,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金(No.3047046730300082);广东省科委重点攻关项目(No.2002C31108);广东省自然科学基金(No.04009360)

摘　　要：目的探讨造影剂声诺维(SonoVue)联合超声辐照人脐血管内皮细胞(HUVEC)时增强型绿色荧光蛋白报告基因质粒(pEGFP)的瞬时转染效率。方法使用连续多普勒超声辐照方式介导pEGFP转染HUVEC,照射条件为频率1.9MHz,TIS0.8,SATA为80.0mW/cm2,时间持续5min,添加或不添加2%SonoVue,pEGFP浓度为50μg/ml。48h后用流式细胞仪检测绿色荧光蛋白(GFP)瞬时转染表达率,并用台盼蓝染色检测细胞活性(生存率)。结果单纯超声辐照组GFP转染率仅为1.5%±0.2%,SonoVue联合超声辐照组为16.1%±1.9%(P<0.001);两组细胞生存率分别为94.1%±2.3%和91.1%±4.1%(P>0.05)。结论SonoVue联合超声辐照条件可增强pEGFP转染HUVEC的效率,而对细胞活性无明显影响,此方法可用于目的基因转染。Objective To investigate the transfection efficiency of plasmid vector coding enhanced green fluorescence protein (pEGFP) in human umbilical vein endothelial cell (HUVEC) by using ultrasound exposure and ultrasound contrast agent (SonoVue).Methods HUVEC and pEGFP (50μg/ml) were exposed to continuous wave Doppler ultrasound (Frequency,1.9 MHz;TIS,0.8;Gain,80%;Ⅰ_SATA ,80.0 mW/cm2) for 5 min,with or without ultrasound contrast agent (2% SonoVue).Forty-eight hours later,the transient expression rate was observed under flow cytometry,and the survival rate of HUVEC was calculated with trypan blue stain.Results The transfection expression rate was 1.5±0.2% in ultrasound exposure group,but increased significantly to 16.1±1.9% after the addition of SonoVue (P<0.001).The survival rates of HUVEC in ultrasound exposure group with and without SonoVue were 94.1±2.3% and 91.1±4.1% respectively (P>0.05).Conclusions The transfection efficiency of pEGFP into HUVEC under ultrasound exposure increases after addition of SonoVue,whereas the survival rate of HUVEC is not influenced.Such a method is useful in the transfection of target gene.

关 键 词：血管内皮细胞 超声辐照 增强绿色荧光蛋白 声诺维 实验研究 SonoVue 增强型绿色荧光蛋白 PEGFP 流式细胞仪检测 HUVEC 报告基因质粒 细胞活性 细胞生存率 转染效率 照射条件 瞬时转染 染色检测 基因转染 造影剂 多普勒 

分 类 号：R346[医药卫生—基础医学]