| Project/Area Number |
07660387
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KUWABARA Mikinori Hokkaido Univ., Graduate School of Vet.Med., Professor, 大学院・獣医学研究科, 教授 (10002081)
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| Co-Investigator(Kenkyū-buntansha) |
INANAMI Osamu Hokkaido Univ., Graduate School of Vet.Med., Associate Professor, 大学院・獣医学研究科, 助教授 (10193559)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1996: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1995: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Nitric oxide / Macrophage / MGD / ESR / Interferon-gamma / Lipopolysaccharide / lactoferrin / spin-trapping / 単球 / 一酸化窒素 / スーパーオキシド / スピントラッピング / 電子スピン共鳴法 / メチルグルカミンジカルバメート |
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| Research Abstract |
In order to investigate nitric oxide (NO) production from macrophage during maturation, peneatrial macrophage was isolated from thioglycolate-stimulated mouse and the production of NO after stimulation by interferon-gamma (INF-gamma), lipopolysaccharide (LPS) and lactoferrin (LF) was measured by the method combining electron-spin-resonance spectroscopy (ESR) and spin-trapping. The complex (MGD-Fe) of Fe^<2+> and N- (Dithiocarbonyl) -N-methylglucamine was used as spin-trapping reagent. Triplet ESR signal due to MGD-Fe-NO,which was produced by the reaction of NO and MGD-Fe, was time-dependently observed in LPS+Inf-gamma-stimulated macrophage. The NO production was also confirmed the increased-responses of NO_2^-+NO_3^-, metabolites of NO,by colorimetric method using Griess reagents. It was also shown that NO synthase detected by Western blotting was induced by these stimulation. Furthermore, LF promoted the NO production and iNOS induction in INF-gamma-stimulated macrophage, whereas LF itself did not produce any NO production in macrophage. These results was suggested that the maturation of macrophage was induced by LF which was produced from neutrophil in inflammation site as well as various cytokines which was produced from various lymphocytes.
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