Development of gene transfer by in vivo electroporation to the vascular wall

我们建立了一种非病毒的动脉基因转移方法，即通过体内电穿孔的方法将质粒DNA导入动脉。电穿孔是通过向细胞或组织施加持续时间短、强度高的电场脉冲，在细胞膜上形成纳米级的瞬变孔洞的物理过程。有效的电穿孔最重要的参数是：刺激电压、脉冲宽度、电流、质粒浓度和电极形状。结果：电压为30、脉冲时间为20ms的电穿孔显著增加了血管壁上基因的表达。最适浓度为400μg/ml。通过体内电穿孔转移的血管的咬痕集中在电极的边缘。血管壁仿真模型和光学标测实验也证实了这一结论。我们比较了单板电极和多板电极在血管壁中的基因表达，单板电极是一个5×1 mm的平板，多板电极是平行的两个平板。综上所述，我们发展了一种利用电穿孔技术进行基因转移的方法，该方法被证明是一种比使用裸DNA更安全有效的方法。然后就有可能在未来处理基因表达的位置或数量。

We developed a nonviral method for arterial gene transfer by in vivo electroporation with plasmid DNA. Electroporation is the physical process of including nanometreaized transient pores in a cell membrane by the application of short duration, high intensity electric field pulses to cells or tissues. The most important parameters for effective electroporation are: the stimulation voltage, the pulse duration, the electric current, plasmid concentration, and the shape of an electrode.Result: Electroporation with 30 voltage and pulse duration of 20 msec resulted in a marked increase of gene expression in vessel wall. The concentration of 400μ g/ml plasmid DNA is most optimal. The Bite of the vessel gene-transferred by in vivo electroporation is focused around the edge of the electrode. The same findings are demonstrated by the simulation model of vessel wall and the experiment of optical mapping. We compare with gene expression in vessel wall between with the monoplate electrode, which is one 5^*1mm plate, and the multiplate electrode, which is two plates in parallel. The gene expression using muitiplate electrode is superior to the one of monoplate electrode.In summary, we developed a method for gene transfer using electroporation, an approach which proved to the safe and more efficient than that using naked DNA. And then it will be possible that the site or the mount of gene expression is handled in the future.