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ANALYSIS OF ENDOGENOUSLY PRODUCED NITRIC OXIDE (NO) EMPLOYING IRON COMPLEXES AS AN NO TRAPPING REAGENT AND PHYSIOLOGICAL STUDY ON NO

Research Project

Project/Area Number 08640781
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 分離・精製・検出法
Research InstitutionINSTITUTE FOR LIFE SUPPORT TECHNOLOGY (ILST), YAMAGATA TECHNOPOLIS FOUNDATION

Principal Investigator

YOSHIMURA Tetsuhiko  ILST,DIVISION OF BIOINORG.CHEM,RES,DIRECTOR, 化学第一研究部, 部長 (70271517)

Co-Investigator(Kenkyū-buntansha) FUJII Satoshi  ILST,DIVISION OF BIOINORG.CHEM,RESEARCHER, 化学第一研究部, 研究員 (80271518)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsNITRIC OXIDE / ENDOGENOUSLY PRODUCED NITRIC OXIDE / TRAP / IRON COMPLEX / ELECTRON SPIN RESONANCE / DITHIOCARBAMATE / N- (DITHIOCARBOXY) SARCOSINE
Research Abstract

Diatomic molecule, nitric oxide (NO) is an inorganic free radical. It has been known as an atomospheric pollutant and a potential hazard. However, it is now widely known as an endogenous molecule with critical physiological roles in the biological processes of vascular smooth muscle relazation, neuronal transmission and macrophage-induced cytotoxicity. 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.
Iron complexes with dithiocarbamate derivatives are noted among the … More 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 measurement. Such dithiocarbamate derivatives include N,N-diethyldithiocarbamate (DETC), N-methyl-D-glucamine dithiocarbamate (MGD), and N-(dithiocarboxy) sarcosine (DTCS). Iron complex with DTCS (Fe-DTCS) and its NO complex (NO-Fe-DTCS) are fairy soluble and stable in aqueous media. Therefore, Fe-DTCS and NO-Fe-DTCS complexes have a potential as a biologically benign, effective NO-trapping reagent and as a water-soluble spin-labeling reagent (or spin probe), respectively. Further, if the EPR signal from endogenously produced or exogenously supplied NO-Fe-DTCS complex is able to measure in the whole body of a live small animal, in vivo EPR imaging of endogenous and exogenous NO would be performed. Such EPR image would permit noninvasive observation of in vivo spatial distribution of NO.
To obtain EPR spectra and image, we used a home built 700-MHz-microwave EPR imaging system, which is composed of an electromagnet attached with a pair of field gradient coils, a data-processing computer, and a 700-MHz-microwace EPR unit.
In this study, we demonstrated the usefulness of iron dithiocarbamates as an NO trapping reagent. Further, we reported the applications of the NO trapping reagent to the detection and imaging of endogenously produced NO.First, we tried the detection of NO generation from porcine aorta endothelial cells. Second, we demonstrated in vivo EPR imaging of endogenously produced NO,trapped by an Fe-DTCS complex, in the abdomen of a live mouse. This experiment has been cariied out in a septic-shock model by administering lipopolysaccharide to mice via intraperitoneal route. The outline of the image thus obtained corresponds to the liver, suggesting the metabolic pathway of NO-Fe-DTCS complex.
We could demonstrate in this study that iron dithiocarbamates as an NO trapping reagent are suitable for in vivo real time detection of NO produced under physiological and pathological conditions. Less

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] 吉村 哲彦: "In vivo ESR imaging of endogenous nitric oxide in Iipopolysaccharide-treated mice" Nature Biotechnology. 14. 992-995 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 横山 秀克: "In vivo ESR-CT imaging of the liver in mice receiving subcutaneous injection of nitric oxide bound iron complex" Magnetic Resonance Imaging. 15. 249-253 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 鈴木 保宏: "The origin of an EPR signal observed in dithiocarbamate-Ioaded tissues:Copper(II)-dithiocarbamate complexes account for the narrow hyperfine lines" Biochimica et Biophysica Acta. 1335. 242-245 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 鈴木 保宏: "In vivo nitric oxide detection in the septic rat brain by electron paramagnetic resonance" Free Radical Research(印刷中). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 藤井 敏司: "In vivo three-dimensional EPR imaging of nitric oxide production from isosorbide dinitrate in mouse" American Journal of Physiology(印刷中). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 吉村 哲彦: "In vivo EPR detection and imaging of endogenously produced and exogenously supplied nitric Oxide" Analytical Sciences. 13(Sup.). 451-454 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 吉村 哲彦: "NOのすべて" 医歯薬出版株式会社(平田結喜緒編), 3-7,30-34 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 吉村 哲彦: "NO-化学と生物-" 学会出版センター(吉村・他編), 8-19,116-125,151-160 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yoshimura, H.Yokoyama, S.Fujii, K.F.Takayama, Oikawa, H.Kamada: "In-vivo EPR imaging of endogenous nitric oxide in lipopolysaccharide-treated mice" Nature Biotechnology. 14. 992-995 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Fujii, T.Yoshimura, H.Kamada: "Nitric oxide trapping efficiencies of water-soluble iron (III) complexes with dithiocarbamate derivatives" Chemistry Letters. 785-786 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Fujii, G.Miyakoda, M.Chihiro, T.Yoshimura, H.Kamada: "Nitric oxide release from porcine aorta endotherial cells measured by electron paramagnetic resonance spectroscopy" Chemistry Letters. 1055-1056 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Yokoyama, S.Fujii, T.Yoshimura, H.Ohya-Nishiguchi, H.Kamada: "In vivo ESR-CT imaging of the liver in mice receiving subcutaneous injection of nitric oxide-bound iron complex" Magnetic Resonance Imaging. 15. 249-253 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Suzuki, S.Fujii, T.Tominaga, T.Yoshimoto, T.Yoshimura, H.Kamada: "The origin of an EPR signal observed in dithiocarbamate-loaded tissues : Copper (II) -dithiocarbamate complexes account for the narrow hyperfine lines" Biochimica et Biophysica Acta. 1335. 242-245 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Fujii, Y.Suzuki, T.Yoshimura, H.Kamada: "In vivo three-dimentional EPR imaging of nitric oxide production from isosorbide dinitrate in mouse" American Journal of Physiology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Suzuki, S.Fujii, Y.Numagami, T.Tominaga, T.Yoshimoto, T.Yoshimura: "In vivo nitric oxide detection in the septic rat brain by electron paramagnetic resonance" Free Radical Research. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yoshimura: "Endogenous nitric oxide, its chemical characterization and the high affinity for metal ions and metalloproteins" CVD Ground Round Series. Vol 3 (Nitric Oxide Grand Round) (Editor-in-Chief, T.Yoshimoto). 1-26 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T,Yoshimura: "In vivo EPR detection and imaging of endogenously produced and exogenously supplied nitric Oxide" Analytical Sciences. 13 (Suppl.). 451-454 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 横山秀克: "In vivo ESR-CT imaging of the liver in mice receiving subcutaneous injection of nitric oxide bound iron complex" Magnetic Resonance Imaging. 15. 249-253 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 鈴木保宏: "The origin of an EPR signal observed in dithiocarbamate-loaded tissues : Copper(II)-dithiocarbamate complexes account for the narrow hyperfine lines" Biochimica et Biophysica Acta. 1335. 242-245 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 藤木敏司: "In vivo three-dimensional EPR imaging of nitric oxide production from isosorbide dinitrate in mouse" American Journal of Physiology(印刷中). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 鈴木保宏: "In vivo nitric oxide detection in the septic rat brain by electron paramagnetic resonance" Free Radical Research(印刷中). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 吉村哲彦: "In vivo EPR detection and imaging of endogenously produced and exogenously supplied nitric oxide" Analytical Sciences. 13(Sup.). 451-454 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 吉村哲彦: "NOと酸素およびチオールとの生体内相互作用" 最新医学・特集「NO:その後の展開」. 52. 919-925 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 吉村哲彦: "In vivo EPR imaging of endogenous nitric oxide in Iipopolysaccharide-treated mice" Nature Biotechnology. 14. 992-995 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 藤井敏司: "Nitric oxide trapping efficiencies of water-soluble iron (III) complexes with dithiocarbamate derivatives" Chemistry Letters. 785-786 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 藤井敏司: "Nitric oxide release from porcine aorta endotherial cells measured by EPR spectroscopy" Chemistry Letters. 1055-1056 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 横山秀克: "In vivo ESR-CT imaging of the liver in mice receiving subcutaneous injection of nitric oxide bound iron complex" Magnetic Resonance Imaging. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 吉村哲彦: "NOとヘムおよび非ヘム鉄タンパク質との反応" 磁気共鳴と医学. 7. 9-13 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 藤井敏司: "ニトロシル鉄錯体をスピンプローブとするL-band ESRイメージング" 磁気共鳴と医学. 7. 169-172 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 吉村哲彦: "NOのすべて" 医歯薬出版株式会社(平田結喜緒編), 3-7,30-34 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 吉村哲彦: "NOー化学と生物ー" 学会出版センター(吉村・他編), 8-19,116-125,151-160 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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