| Project/Area Number |
04454543
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | KOBE UNIVERSITY |
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Principal Investigator |
OKUMURA Katsuhiko Depart.Hospital Pharmacy Professor, 医学部・付属病院, 教授 (60025707)
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| Co-Investigator(Kenkyū-buntansha) |
IWAKAWA Seigo Kobe Pharmaceutical Univ.Depart.Pharmaceutical Science Associate Professor, 薬剤学教室, 助教授 (50168548)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1992: ¥3,900,000 (Direct Cost: ¥3,900,000)
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| Keywords | Superoxide dismutase / Gene therapy / SOD cDNA / Culture cell / Superoxide / Secretion / Non-secretion / Catalase / SOD cDNA / 形質転換細胞 / スーパーオキサイドジスムターゼ / ラット / cDNA / 呼吸窮迫症候群 / PCR |
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| Research Abstract |
Human Cu, Zn-SOD (superoxide dismutase) cDNA was inserted in eukaryotic expression vectors (pRc/RSV and pRc/CMV), and then those vectors were transfected in rat skin fibroblast cell (FR cell), rat skin primary culture cell, and rat lung epitherial cell (L2 cell) with Lipofection method. Each SOD concentration in transfected cells was increased. These transfected cells resisted to a treatment of superoxide synthesized with Paraquart and xanthin/xanthin oxidase methods induced cell killing. After treatment of superoxide, the lipid peroxide concentration in these transfected cells was less than that of normal cells.
Expression vector (IL-SOD CMV) was constructed containing human Cu, Zn-SOD cDNA fused in a frame to a leader sequence of human interleukin-2 (IL-2) gene under the control of a viral promoter. FR cell and L2 cell transfected with this vector synthesized and secreted SOD in culture medium. These transfected cells resisted to a treatment of superoxide induced cell killing. The lipid peroxide concentration in these transfected cells was less than normal cells after treatment of superoxide.
These results show that the transfection of SOD cDNA in cell was effective in the superoxide induced cytotoxicity.
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