裸DNA肌肉注射不同注射方法对标记基因转染效率的影响  

The Effect of Different Injection Techniques of Naked DNA on Marker Gene Transfer Expression

作　　者：文立[1] 陈立[2] 贺晨霞[2] 陆爱云 曾凡辉 卢大儒[2] 薛京伦[2] 

机构地区：[1]天津体育学院运动医学研究所,天津300381 [2]复旦大学遗传研究所 [3]上海体育学院

出　　处：《中国运动医学杂志》2003年第6期580-583,602,共5页Chinese Journal of Sports Medicine

摘　　要：在肌肉损伤基因治疗应用研究中 ,裸DNA直接注射法具有安全可靠 (不会引起炎症反应和导致插入突变 )、制作工艺简单、操作简便 (与传统的注射方法非常相近 )、可重复注射等优点。以往研究表明 ,肌肉DNA注射基因转移的注射技术是显著影响表达水平的一个重要参数。因此 ,本研究比较了直接注射、注射后保持和缩进式注射三种不同注射方法对基因转移效率的影响 ,进行了不同注射方法的机理探讨。实验结果显示 ,缩进式注射方法转染效率最高 ,直接注射和注射后保持两种方法间无显著性差异。结果提示 ,质粒不是通过细胞膜的破损进入细胞 ,可能是“受体介导”或“T管和内陷”作用的结果。这一结果支持“T管和细胞内陷说”和“受体介导说”。Introduction of an exogenous gene into cells in the form of naked DNA has many obvious advantages. It is easy to obtain significant amounts of DNA. In addition, the plasmid has been shown to remain unintegrated, or extrachromosomal. Thus, plasmid-based gene therapy by intramuscular injection may be safe, cost-effective and have high patient compliance due to the acceptability of intramuscular injection. It has been showed that injection techniques are an important influence facet. Three injection techniques, i.e. “longitudinal injection', “inject and keep needle a while', “retraction/injection', have been compared in this study. The results showed that using the “retraction/injection' method, the levels of β-gal expression in mouse rectus femoris muscle were greater than other methods, and there is no significant difference between “longitudinal injection' and “inject and keep needle a while' method. These results indicated that membrane disruption is not essential process in gene transfer to muscle, implied that DNA uptakes mechanism based on receptor-mediated or T tubules and caveolae uptake.

关 键 词：裸DNA肌肉注射 注射方法 基因转染效率 影响 骨骼肌 :肌肉创伤 

分 类 号：R87[医药卫生—运动医学]