Basic Research for HGF Gene Transduction to Kidney

点击翻译按钮获取中文摘要

HGF gene induction using HVJ-liposome was examined. To look for the most efficient method, intra-muscular, intra-ureteral or intra-arterial injection were tested as an administration route, anionic or AVE (artificial viral envelope) were tested in liposome type, and PUC SR α-HGF, EBNA-HGF or pcDNA3-HGF were tested as a vector. Animals used were 5 week-old Wister rat. Intra-muscular injection was made into bilateral femur 4-6 times. Intra-ureteral injection was done after clamping the renal artery and vein and the ureter was ligated 5 minutes later. Intra-arterial injection was done via renal artery after clamping the renal artery and vein and the clamp was removed 5 minutes later. HGF expression was evaluated by ELISA using anti-human HGF antibody 3 days after the injection. HVJ-liposome has a lipid membrane associated with HVJ virus on outer layer and includes induction gene in it. When it fuses with a cell, the gene is introduced into the cell. Among the several kinds of liposome, anionic liposome and AVE were employed. Anionic liposome is negatively charged and is ordinarily used for gene induction in vivo. AVE, which has been developed recently, resembles the viral membrane. Three kinds of vector were used which have different promoters. As a result, there was no significant difference in HGF expression between treated and contralateral kidney by intra-ureteral or intra-arterial. Intra-muscular method was expected to avoid the influence of the surgical damages to the kidney. Finally, HGF gene induction was most efficient when AVE liposome was used with pcDNA3-HGF as a vector. The effect of HGF on renal damages will be investigated according to this procedure in the future.