Maruyama H, Ataka K, Higuchi N, Sakamoto F, Gejyo F, Miyazaki J
Department of Medicine (II), Niigata University School of Medicine, Niigata, Japan.
Gene Ther. 2001 Dec;8(23):1808-12. doi: 10.1038/sj.gt.3301604.
The skin is an important target for gene transfer because of its easy accessibility. Using plasmid DNA expressing rat erythropoietin (pCAGGS-Epo) as the vector, we previously demonstrated long-term Epo delivery in rats by muscle-targeted gene transfer using in vivo electroporation. Here we examined whether this electroporation approach could be applied to gene delivery in rat skin. To optimize gene transfer, we tested the efficiency of skin-targeted Epo gene transfer with three types of electrodes at three different electrode voltages. Each rat was injected intradermally with a total of 800 microg of pCAGGS-Epo, in the abdominal area. Plate-and-fork-type electrodes were effective for Epo delivery by skin-targeted gene transfer at low voltages (12 approximately 24 V). The vector-derived Epo mRNA was expressed only at the DNA injection site. The Epo gene was expressed in a dose-dependent manner, the expression persisted for 7 weeks, and hematocrit levels were increased for 11 weeks. Skin injection with pCAGGS-lacZ showed lacZ gene expression in the epidermis on day 1 after injection and in the subcutaneous muscle layer on day 7. Slight skin damage due to the gene transfer procedure was evident on day 1, but absent by day 7. These results demonstrate that skin-targeted pCAGGS-Epo transfer by in vivo electroporation at low voltage is a useful procedure for the short-term delivery of Epo.
由于皮肤易于接近，所以它是基因转移的一个重要靶标。我们先前使用表达大鼠促红细胞生成素的质粒DNA（pCAGGS-Epo）作为载体，通过体内电穿孔的肌肉靶向基因转移，在大鼠中证明了促红细胞生成素的长期递送。在此，我们研究了这种电穿孔方法是否可应用于大鼠皮肤的基因递送。为了优化基因转移，我们在三种不同的电极电压下，用三种类型的电极测试了皮肤靶向促红细胞生成素基因转移的效率。每只大鼠在腹部区域皮内注射总共800微克的pCAGGS-Epo。平板叉型电极在低电压（12至24伏）下通过皮肤靶向基因转移对促红细胞生成素的递送有效。载体衍生的促红细胞生成素mRNA仅在DNA注射部位表达。促红细胞生成素基因以剂量依赖性方式表达，表达持续7周，血细胞比容水平升高11周。注射pCAGGS-lacZ后，在第1天表皮和第7天皮下肌肉层显示出lacZ基因表达。基因转移过程导致的轻微皮肤损伤在第1天明显，但在第7天消失。这些结果表明，通过低电压体内电穿孔进行皮肤靶向pCAGGS-Epo转移是促红细胞生成素短期递送的一种有用方法。
