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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pEGFPC1基因转导,HepG2细胞以1MHz1.5W/cm2超声波基因转导效果最好;小鼠胫前肌以1MHz3W/cm2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。
Objective To investigate the feasibility of SonoVue ultrasound-mediated microbubble ultrasound contrast agent gene transduction in cultured cells and in vivo muscle tissue and to study the relationship between different ultrasonic parameters and gene dosage and the efficiency of transgene. Methods The transduction of enhanced green fluorescent protein (EGFP) reporter plasmid in the right tibiae of human hepatocellular carcinoma cell line HepG2 and BALB / c mice was mediated by ultrasound and acoustic contrast agent SonoVue with different intensities and different doses. The frozen or frozen sections After the observation under a fluorescence microscope, and calculate the transduction efficiency. Results Ultrasound combined with microbubble SonoVue could mediate the transduction of pEGFPC1 gene in HepG2 cells and mouse anterior tibial muscle in vitro. The transduction efficiency of HepG2 cells with 1MHz 1.5W / cm2 ultrasound was the best. The anterior tibial muscle of mice 1MHz3W / cm2 ultrasound transgene best. Conclusion Ultrasound combined with microbubbles SonoVue can mediate gene transduction in cultured cells and in vivo muscle tissue. 1 MHz ultrasound is the ideal frequency of transgene.