超声/微气泡SonoVue介导体内和体外基因转导的研究

目的初步探讨超声联合微气泡超声造影剂SonoVue介导体外培养细胞和体内肌肉组织基因转导的可行性，并研究不同超声波参数和基因用量与转基因效率之间的关系。方法利用不同强度和不同剂量超声波和声学造影剂SonoVue，介导人肝癌细胞株HepG2和BALB／c小鼠右侧胫前肌内增强型绿色荧光蛋白报告质粒的转导，分别直接或组织冰冻切片后在荧光显微镜下观察，并计算转导效率。结果超声联合微气泡造影剂SonoVue可介导体外培养的HepG2细胞和小鼠胫前肌内pEGFP-C1基因转导，HepG2细胞以1MHz 1．5W／cm^2超声波基因转导效果最好；小鼠胫前肌以1MHz3W／cm^2的超声波转基因效果最好。结论超声联合微气泡造影剂SonoVue可介导体外培养细胞和体内肌肉组织基因转导，1MHz超声波是转基因的理想频率。

Study of in vitro and in vivo Gene Transfection Mediated by Ultrasound and Microbubble SonoVue

Objective To investigate the feasibility and the efficacy of gene transfection to cultured cells and mouse muscles, mediated by ultrasound combined with a new ultrasound contrast agent SonoVue. The efficiency of gene expression with regard to different parameters of ultrasound and different DNA dosages was also studied. Methods Human liver cancer cell line HepG2 was used in the in vitro gene transfection experiment. While intramuscular injection of plasmid DNA combined with SonoVue solution was used in the in vivo experiment. Results pEGFP-C1 plasmids could be readily transfected to HepG2 cells in vitro and to mouse tibialis anterior muscles in vivo. One MHz ultrasound was ideal both in vitro and in vivo, but a more powerful ultrasound was needed in vivo (3 W/cm 2) than in vitro (1.5 W/cm 2). Conclusion Ultrasound combined with SonoVue could promote gene transfection to cultured cells and mouse muscles. In both experiments 1 MHz was an ideal ultrasound frequency.