Enhanced gene delivery into skeletal muscles with ultrasound and microbubble techniques
Enhanced gene delivery into skeletal muscles with ultrasound and microbubble techniques
复制标题
通过超声波和微泡技术增强基因向骨骼肌的传递。
DOI：
10.1016/j.acra.2005.11.003
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发表时间：
2006-03-01
影响因子：
4.8
通讯作者：
Schutt, CE
中科院分区：
文献类型：
--
作者：
Zhang, QX;Wang, ZG;Schutt, CE
理由和目标。本实验旨在探讨超声微泡对体外基因结构的影响及体内绿色荧光蛋白（GFP）质粒转染骨骼肌的作用。通过建立大鼠缺血后肢模型，在体内研究超声介导的微泡破坏对血管内皮生长因子（VEGF）基因转染骨骼肌的影响。超声辐照微泡与GFP质粒的混合物。凝胶电泳检测超声波和微泡对GFP质粒的影响。用于体内实验。在将微泡直接注射到Wistar大鼠的后肢中之后，对后肢施加超声照射。处理后立即取骨骼肌观察显微结构。应用超声和微泡技术研究了GFP质粒DNA在大鼠骨骼肌中的转移率。进一步应用裸VEGF质粒在大鼠脑缺血模型上研究其血管生成的可行性。质粒DNA凝胶电泳显示各组间无差异。通过对骨骼肌的苏木精-伊红染色照片的研究发现，注射微泡后超声照射骨骼肌，可引起红细胞渗出，而对肌纤维的显微结构无影响。体内实验表明，超声微泡可以促进质粒DNA向骨骼肌的转移。超声介导微泡技术为基因治疗提供了一种有效的无创方法。
Rationale and Objectives. This experiment was directed to explore the effects of ultrasound microbubbles on gene structure in vitro and green fluorescent protein (GFP) plasmid transfer into skeletal muscles in vivo. By establishing a rat ischemic hind limb model, the effects of ultrasound-mediated microbubble destruction on vascular endothelial growth factor (VEGF) gene transfection to skeletal muscles were also studied in vivo.Materials and Methods. Ultrasound irradiation was applied on the mixture of microbubbles and GFP plasmid in vitro. Gel electrophoresis was used to detect the effects of ultrasound and microbubbles on GFP plasmid. For in vivo experiments. ultrasound irradiation was applied on the hind limb after directly injecting microbubbles into the hind limb of Wistar rats. Directly after treatment, the skeletal muscles were harvested to observe the microstructure. We also studied the transfer rate of GFP plasmid DNA into the skeletal muscles of rats by applying ultrasound and microbubble technique. Furthermore, a naked VEGF plasmid was applied to study the feasibility of angiogenesis by using rats ischemia models.Results. Gel electrophoresis of plasmid DNA showed that there was no difference between the groups. By studying the hematoxylin and eosin stained pictures of the skeletal muscles, we found that ultrasound irradiation of skeletal muscle after injection of microbubbles could cause the exudation of the red blood cells, whereas it had no effects on the microstructure of muscle fibers. In vivo experiments showed that an ultrasound microbubble could enhance the transfer of plasmid DNA to the skeletal muscles.Conclusions. The ultrasound-mediated microbubble technique provides an effective noninvasive method for gene therapy.