1999 Fiscal Year Final Research Report Summary
ESR ANALYSIS AND IN VIVO ACTION OF ENDOGENOUS NITRIC OXIDE
| Project/Area Number |
10044107
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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 |
分離・精製・検出法
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| Research Institution | INSTITUTE FOR LIFE SUPPORT TECHNOLOGY (ILST) |
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Principal Investigator |
YOSHIMURA Tetsuhiko ILST, DIVISION OF BIOINORGANIC CHEMISTRY, RESEARCH DIRECTOR, 生物ラジカル研究所・化学第一研究部, 部長 (70271517)
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| Co-Investigator(Kenkyū-buntansha) |
FUJII Satoshi ILST, DIVISION OF BIOINORGANIC CHEMISTRY, RESEARCHER, 生物ラジカル研究所・化学第一研究部, 研究員 (80271518)
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| Project Period (FY) |
1998 – 1999
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| Keywords | NITRIC OXIDE / ENDOGENOUSLY PRODUCED NITRIC OXIDE / SPIN TRAP / IRON COMPLEX / ELECTRON SPIN RESONANCE / DITHIOCARBAMATE / N-(DITHIOCARBOXY)SARCOSINE / PHENYL BUTYLNITRONE |
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| Research Abstract |
A variety of information about physiological and pathological actions of nitric oxide has been found very useful for the development of drugs as Viagra. To elucidate a variety of actions of endogenous NO, information concerning the quantities and distributions of NO in cells, tissues, and organs is essential. However, it is rather difficult to determine quantities and distribution due to the very small concentration of NO formed in vivo and the very short half-life of NO in living system. As one of the analytical methods to overcome these difficulties, spin-trapping technique combined with electron paramagnetic resonance (EPR) spectroscopy has been used for the determination of unstable free radicals in vitro and in vivo.
1. Iron complexes with dithiocarbamate derivatives are noted among the spin trapping reagents for NO because NO has a high affinity for the iron complexes and resultant NO-bound iron complexes exhibit an intense three-line signal at room temperature on in vitro EPR mea
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surement. N-(dithiocarboxy) sarcosine (DTCS) is a derivative of dithiocarbamate. We recently reported that iron complex with DTCS (Fe-DTCS) and its NO complex (NO-Fe-DTCS) are fairly soluble and stable in aqueous media and Fe-DTCS complex have a potential as a biologically benign, effective NO-trapping reagent. In this study, we measured endogenously produced nitric oxide by employing EPR spin-trapping technique and iron complex with dithiocarbamate as an NO trapping agent. First, we tried in vivo detection of nitric oxide in the brain of living rat during experimental sepsis and meningitis. It was demonstrated that NO is generated in vivo by inducible NO synthase in the brain during sepsis and meningitis. Second, we measured NO production from a nitrovasodilator, isosorbide dinitrate, in mice by in vivo EPR spectroscopy and obtained three-dimensional EPR images of the upper abdominal region.
2. This project was started as a joint research of International Scientific Research Program in 1998. We collaborated with Dr. Yashige Kotate of Oklahoma Medical Research Foundation in USA. In this joint research, we investigated the pharmacological activities of a spin-trapping agent, phenyl-N-tert-butyl nitrone (PBN) in various biological systems. The results obtained suggested that PBN has the function like a non-steroidal anti-inflammatory drug and it reduces the nitric oxide production in the rat brain of meningitis model. Less
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