超声微泡造影剂介导小鼠骨骼肌基因转染实验研究

目的探讨微泡造影剂在超声作用下是否可增加小鼠骨骼肌基因转染效率.方法 40只昆明小鼠随机分为4组,每组10只,第一组:在胫前肌注射造影剂与绿色荧光蛋白(GFP)质粒的混合溶液;第二组:注射与第一组相同的混合溶液后立即加超声辐照;第三组:注射GFP;第四组:在注射GFP后立即用超声辐照.7天后取小鼠胫前肌观察绿色荧光蛋白的表达情况.结果第一组与第二组有较多GFP表达,部分肌纤维绿色荧光较明亮,部分较暗淡;第三组和第四组GFP表达量较少.第一组与其余各组间的差异有显著性意义,P＜0.05;第二组与其余各组间的差异有显著性意义,P＜0.05;第三组与第四组间的差异无显著性意义,P＞0.05.结论超声微泡造影剂在超声作用下可明显增强小鼠骨骼肌的基因转染效率;未加超声波作用时,直接肌注携基因的超声微泡造影剂亦可增加小鼠骨骼肌的基因转染效率.

Microbubble mediating plasmid DNA transfection into mouse skeletal muscle by using ultrasound

Objective To explore whether the microbubble of ultrasonic contrast agent destructing by ultrasound can improve the efficiency of gene transfection in skeletal muscle of mouse in vivo. Methods Forty male mice were randomly divided into 4 groups. Microbubbles attached with the naked plasmid DNA of green fluorescent protein (GFP) were infused into the pretibial muscle of mice (group one and two) with or without destructing microbubbles by ultrasound immediately. The naked plasmid DNA of GFP without microbubbles was infused into the pretibial muscle of mice (group three and four) with or without destructing microbubbles by ultrasound immediately. Mice were all killed 7 days after plasmid DNA infusion and the tissue samples of pretibial muscle were obtained to observe the expression of GFP. Results In the first and the second group the GFP in much muscle expressed, some fluorescence in the muscle was bright, some was dark. In the third and the forth group the expression of GFP was much lower than that in the first and the second group. The difference between the first and the other groups was significant, P<0.05.The difference between the second and the other groups was significant, P< 0.05. The difference between the third and the forth group was not significant, P>0.05. Conclusion The microbubble of ultrasonic contrast agent destructing by ultrasound could significantly improve the efficiency of gene transfection in skeletal muscle of mouse in vivo,and the microbubble of ultrasonic contrast agent without destructing by ultrasound also could improve the efficiency of gene transfection in skeletal muscle of mouse in vivo.