| Project/Area Number |
09470346
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | Osaka University |
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Principal Investigator |
MATSUMIYA Kiyomi Graduate School of Medicine, Osaka University Instructor, 医学系研究科, 講師 (90243237)
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| Co-Investigator(Kenkyū-buntansha) |
KITAMURA Masaya Graduate School of Medicine, Osaka University Assistant Professor, 医学系研究科, 助手 (70273688)
TAKAHARA Shiro Graduate School of Medicine, Osaka University Associate Professor, 医学系研究科, 助教授 (70179547)
OKUYAMA Akihiko Graduate School of Medicine, Osaka University Professor, 医学系研究科, 教授 (20093388)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Hepatocyte Growth Factor / gene induction / HVJ-liposome / kidney / rat / protective effect on renal function / 肝細胞腎殖因子(HGF) / 腎不全 / 腎阻血 / 5 / 6腎摘除 / リポゾーム法 / HGF / 腎再生作用 / HVJリポゾーム / SHRラット |
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| Research Abstract |
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.
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