纳米微泡介导GFP转染小鼠骨骼肌细胞的实验研究  

Experiment of nanobubbles-mediated plasmid green fluorescent protein{GFP） transfer into skeletal muscle in mice

作　　者：王兴华[1] 石颖[1] 乔英艳[1] 王芹秀[1] 郭艳琴[1] 雷成功[1] 

机构地区：[1]山西医科大学第二临床医学院超声科,太原030001

出　　处：《中华超声影像学杂志》2012年第6期529-532,共4页Chinese Journal of Ultrasonography

摘　　要：目的探讨两种纳米微泡（白蛋白外膜和磷脂外膜）增强基因转染小鼠骨骼肌的作用。方法以正常C57810小鼠胫前肌为研究对象，目的基因GFP与微泡混合注入小鼠胫前肌，一侧胫前肌经超声（1MHz脉冲波，脉冲重复频率为100Hz，20％工作周期，空间峰值时间峰值声强为2W／cm^2，辐照作用时间30s），另一侧胫前肌不经超声辐照。观察白蛋白纳米微泡及磷脂纳米微泡增强骨骼肌细胞GFP转染水平的作用。1周后处死小鼠，荧光显微镜观察发出绿色荧光者为GFP阳性肌纤维细胞，计数最大GFP阳性肌纤维细胞数，作为GFP基因转染效率指标。结果①白蛋白纳米微泡组和白蛋白纳米微泡＋超声组最大GFP阳性肌纤维数显著高于阴性对照组（P〈0．05），显著低于阳性对照组（P〈O．05）。②磷脂纳米微泡组与阴性对照组及阳性对照组比较，最大GFP阳性肌纤维数差异均无统计学意义（P〉0．05）。磷脂纳米微泡＋超声组最大GFP阳性肌纤维数较阴性对照组显著增高（P〈0．05）；磷脂纳米微泡＋超声组与阳性对照组最大GFP阳性肌纤维数差异无统计学意义（P〉0．05）。结论纳米微泡可增强基因转染骨骼肌细胞效率，白蛋白含氟化气体纳米微泡具有发展潜力。Objective To explore the effect of nanoscale bubbles transfering gene in skeletal muscle cells in mice. Methods Plasmid DNA encoding green fluorescent protein （GFP） was mixed with bubbles dissolved in saline and injected into the tibialis anterior （TA） muscle of C57B10 mice with and without ultrasound （US）. The ultrasound frequency was 1 MHz and the pulse repetition frequency was 100 Hz with 20o//oo duty cycle. The spatial peak temporal peak intensity （ISPTP） power level was 2 W/cm2. The entire treatment period was 30 seconds. The efficiencies of GFP transgene expression were determined under different experimental conditions. Mice were sacrificed 1 week after plasmid DNA injection. Fibres with fluorescence green signals were determined as GFP-positive fibres by fluorescence microscopy. Readout was performed on the section with the maximum number of transfected fibers. Results 1 ） Albumin nanobubbles：in the conditon of with or without ultrasound,albumin nanobubbles had significantly increased gene expression compared with negative control （ P 〈 0.05）, but significantly decreased gene expression compared with positive control （ P 〈 0.05）. 2） Phospholipid nanobubbles.. In the conditon of without ultrasound, there were no significant differences compared with negative and positive control （ P 〉0.05）. In the conditon of with ultrasound, phospholipid nanobubbles had significantly increased gene expression compared with negative control （ P d0.05）. No significant difference was observed between phospholipid nanobubbles and positive control （ P 〉0.05）. Conclusions Nanobubbles could enhance gene transfection efficiency for skeletal muscle fibres in mice. Nanobubbles with perfluoropropane gas and albumin have potentiality in gene-enhanced transfection.

关 键 词：超声检查 微气泡 纳米技术 转染 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]