几种共聚物对小鼠骨骼肌超声介导基因转染表达的影响  

The functions of several pluronic block copolymers in the efficiency of ultrasound-induced gene transduction in skeletal muscle of mice in vivo

作　　者：陈云超[1] 黄道中[1] 李开艳[1] 王志辉[1] 洪恺[1] 王芬[1] 张青萍[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院超声影像科,武汉430030

出　　处：《中华超声影像学杂志》2008年第1期77-80,共4页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金面上项目（30670620）;教育部留学回国人员启动基金（教外司留2005-383）

摘　　要：目的探讨超声和共聚物在质粒DNA对小鼠骨骼肌基因转染中的影响。方法应用三种水溶性不同的共聚物F127，L61和P85，与DNA混合后直接小鼠胫前肌注射，并超声辐照。超声频率为1MHz，脉冲重复频率100Hz，占空系数为20％，用3W/cm^2声强辐照1min。实验1周后取出胫前肌并快速冰冻切片，荧光显微镜计数表达GFP的转染肌纤维数，苏木精和伊红染色评价肌肉损伤情况。结果三种共聚物均可显著提高质粒DNA的基因转染（P〈0．01），P85的基因转染效率高于其他两种共聚物（P〈0．05）。超声虽对单纯质粒、F127和L61组的基因转染无显著促进作用，但与P85有协同作用，可显著提高P85组的基因转染效率（P〈0．01）。在给定超声剂量和共聚物浓度下，二者在提高质粒DNA基因转染的同时，并不显著增加对组织的损伤作用。结论共聚物P85不但可显著提高质粒DNA在小鼠骨骼肌的基因转染，且与超声具有协同作用。Objective To explore the effect of ultrasound and pluronics in plasmid DNA skeletal muscle gene transduction in vivo. Methyls Plasmid encoding green fluorescent protein （GFP）, which conjugated with three different pluronics,F127, L61 and P85, was injected into the tibialis anterior（TA） muscle of mice with and without adjunct ultrasound （1 MHz,3 W/cm^2 1min,20% duty cycle）. Mice were killed 1 week after injection. The TA muscles were removed and snap-frozen immediately in isopentane cooled by liquid nitrogen and sections 7 μm thick were cut at intervals. One set of sections mounted with DAPI were used to assess the transfection efficiency by counting the number of GFP-positive fibers under fluorescence microscopy,and the other set of sections were stained with haematoxylin and eosin to assess the tissue damage area. Results The three pluronics significantly enhanced the plasmid DNA skeletal muscle gene delivery in vivo（P〈0.01 ）, especially the P85, which induced gene transduction efficiency higher than those of the other pluronics （P〈0.05）. The ultrasound showed synergistic effect to enhance the gene delivery efficiency with P85 （P〈0.01）, but the ultrasound could not enhance the gene transduction efficiency of plasmid DNA only, F127 and L61 groups. The ultrasound and the pluronics, under the given energy and concentrations,did not increase the tissue damage during enhancing plasmid DNA transduction. Conclusions P85 displays significantly synergistic effect with ultrasound to enhance plasmid DNA transduction in skeletal muscle of mice in vivo.

关 键 词：超声检查 基因转移 肌 骨骼 聚合物 

分 类 号：R346[医药卫生—基础医学]