ESR ANALYSIS AND IN VIVO ACTION OF ENDOGENOUS NITRIC OXIDE

内源性一氧化氮的 ESR 分析和体内作用

• 批准号: 10044107
• 负责人: YOSHIMURA Tetsuhiko
• 金额: $ 2.18万
• 依托单位: INSTITUTE FOR LIFE SUPPORT TECHNOLOGY (ILST)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10044107/
• 关键词: NITRIC OXIDE; ENDOGENOUSLY PRODUCED NITRIC OXIDE; SPIN TRAP; IRON COMPLEX; ELECTRON SPIN RESONANCE; DITHIOCARBAMATE; N-(DITHIOCARBOXY)SARCOSINE; PHENYL BUTYLNITRONE; フェニルブチルニトロン; 一酸化窒素(NO); 電子スピン共鳴法; NOトラップ試薬; NFkB

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 … More 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

关于一氧化氮的生理和病理作用的各种信息已被发现对药物如伟哥的开发非常有用。为了阐明内源性NO的各种作用，有关NO在细胞、组织和器官中的数量和分布的信息是必不可少的。然而，由于NO在体内形成的浓度很低，并且在生命系统中的半衰期很短，因此很难确定其数量和分布。作为克服这些困难的分析方法之一，自旋捕集技术结合电子顺磁共振（EPR）谱已被用于体内外不稳定自由基的测定.铁与二硫代氨基甲酸酯衍生物的络合物在用于NO的自旋捕获试剂中被注意到，因为NO对铁络合物具有高亲和力，并且所得的NO结合的铁络合物在室温下在体外EPR测量中显示出强烈的三线信号。 ...更多信息 保证N-（二硫代羧基）肌氨酸（DTCS）是二硫代氨基甲酸酯的衍生物。铁与DTCS的配合物（Fe-DTCS）及其与NO的配合物（NO-Fe-DTCS）在水溶液中具有良好的溶解性和稳定性，Fe-DTCS配合物具有良好的生物活性和捕集NO的能力。在这项研究中，我们测量内源性产生的一氧化氮，采用EPR自旋捕获技术和铁配合物与二硫代氨基甲酸作为NO捕获剂。首先，我们尝试了在体检测实验性脓毒症和脑膜炎大鼠脑组织中一氧化氮的变化。已经证明，在脓毒症和脑膜炎期间，NO通过脑中的诱导型NO合酶在体内产生。其次，我们通过在体EPR光谱技术测量了硝基血管扩张剂硝酸异山梨酯在小鼠体内产生NO的情况，并获得了上腹部区域的三维EPR图像.该项目于1998年作为国际科学研究计划的一项联合研究开始。我们与美国俄克拉荷马州医学研究基金会的Yashige Kotate博士合作。在这项联合研究中，我们研究了自旋捕获剂苯基-N-叔丁基硝酮（PBN）在各种生物系统中的药理活性。结果表明，PBN具有非甾体抗炎药的作用，能降低脑膜炎模型大鼠脑内一氧化氮的产生。少

项目成果

Y. Suzuki: "Direct evidence of in vivo nitric oxide production and inducible nitric oxide synthase mRNA expression in the brain of living rat during experimental meningitis"Journal of Cerebral Blood Flow and Metabolism. 1335. 242-245 (1997)

Y. Suzuki：“实验性脑膜炎期间活体大鼠大脑中一氧化氮产生和诱导型一氧化氮合酶 mRNA 表达的直接证据”《脑血流与代谢杂志》。

Y.Kotake, H.Sang, T.Miyajima, G.L.Wallis: "Inhibition of NF-κB, iNOS mRNA, COX2 mRNA, and COX catalytic activity by phenyl-N-tert-butylnitrone (PBN)"Biochem. Biophys. Acta. 1448. 77-84 (1998)

Y.Kotake、H.Sang、T.Miyajima、G.L.Wallis：“苯基-N-叔丁基硝酮 (PBN) 对 NF-κB、iNOS mRNA、COX2 mRNA 和 COX 催化活性的抑制”Biochem。 1448. 77-84 (1998)

S.Fujii, Y.Suzuki, T.Yoshimura, H.Kamada: "In vivo three-dimensional EPR imaging of nitric oxide production from isosorbide dinitrate in mouse"Am. J. Physiol.. 274. G857-G862 (1998)

S.Fujii、Y.Suzuki、T.Yoshimura、H.Kamada：“小鼠体内硝酸异山梨酯产生一氧化氮的体内三维 EPR 成像”Am。

S. Fujii: "In vivo three-dimensional EPR imaging of nitric oxide production from isosorbide dinitrate in mouse"American Journal of Physiology. 274. G857-G862 (1998)

S. Fujii：“小鼠体内硝酸异山梨醇产生一氧化氮的体内三维 EPR 成像”美国生理学杂志。

Y.Kotake, D.R.Moore, H.Sang, L.A.Reinke: "Continuous monitoring of in vivo nitric oxide formation using EPR analysis in biliary flow"Nitric Oxide : Biol. Chem.. 3. 114-122 (1999)

Y.Kotake、D.R.Moore、H.Sang、L.A.Reinke：“使用胆汁流中的 EPR 分析连续监测体内一氧化氮的形成”一氧化氮：Biol。