Matsuno Y, Iwata H, Umeda Y, Takagi H, Mori Y, Kosugi A, Matsumoto K, Nakamura T, Hirose H
First Department of Surgery, Gifu University School of Medicine, Gifu, Japan.
Gene Ther. 2003 Sep;10(18):1559-66. doi: 10.1038/sj.gt.3302052.
Although a variety of gene transfer methods to the liver have been designed, there are some problems such as the transfection efficiency and safety. In the present study, we developed a modified method of gene transfer into the liver by infusion of plasmid DNA via the portal vein followed by electroporation. After green fluorescence protein gene transfer, transgene expressions were detected in 24 h, and then maximally at 3 days, and persisted for 3 weeks. Histological analysis revealed that very mild tissue damage was induced in the liver to which electroporation was applied. In the second study, human hepatocyte growth factor (HGF) was more detected in the liver injected with 500 microg of human HGF gene than 100 microg of human HGF gene. However, serum HGF did not increase with 100 or 500 microg of human HGF gene. Moreover, 500 microg of HGF gene transfer into the liver by using this method could achieve the long survival of all dimethylnitrosamine-treated rats and attenuate the fibrous regions in the liver. These results suggest that HGF gene transfer into the liver via the portal vein using electroporation might be one of the useful methods for the treatment of various liver diseases.
尽管已经设计出多种将基因导入肝脏的方法,但仍存在一些问题,如转染效率和安全性。在本研究中,我们开发了一种改良的基因导入肝脏的方法,即通过门静脉注入质粒DNA后进行电穿孔。绿色荧光蛋白基因导入后,在24小时检测到转基因表达,然后在3天时达到最大值,并持续3周。组织学分析显示,接受电穿孔的肝脏仅出现非常轻微的组织损伤。在第二项研究中,注射500微克人肝细胞生长因子(HGF)基因的肝脏中检测到的人HGF比注射100微克人HGF基因的肝脏更多。然而,注射100微克或500微克人HGF基因后血清HGF并未升高。此外,使用该方法将500微克HGF基因导入肝脏可使所有经二甲基亚硝胺处理的大鼠长期存活,并减轻肝脏中的纤维化区域。这些结果表明,通过门静脉电穿孔将HGF基因导入肝脏可能是治疗各种肝脏疾病的有效方法之一。