1993 Fiscal Year Annual Research Report

ESR法による細胞内酸素濃度測定法の開発

Research Project

Project/Area Number	03557098
Research Institution	The University of Tokyo
Principal Investigator	桐野豊東京大学, 薬学部, 教授 (10012668)
Co-Investigator(Kenkyū-buntansha)	嶋本典夫武田薬品工業, 創薬研究本部, 室長内海英雄昭和大学, 薬学部, 助教授 (20101694) 小沢俊彦放射線医学総合研究所, 薬理化学, 部長 (40160858)
Keywords	生体内酸素 / ESR / ニトロキシドラジカル / 活性炭 / 常磁性 / L-バンドESR
Research Abstract	安定常磁性種をプローブとするESR法による生体内酸素濃度測定法を確立することを目的としている。 (1)安定ニトロキシドラジカル、活性炭、fusiniteの3種について、プローブとしての適格性を比較検討した。後2者は安定性やシグナルの大きさ、酸素濃度に対する感受性は大きいものの、酸素濃度変化に追随する速度がきわめて遅い(平衡に達するのが遅い)という致命的な欠点がある。これに対し、ニトロキシドは細胞内では酸化還元系酵素により常磁性を失うという欠点はあるものの、酸素濃度変化に従って速やかにその線幅を変化させるので、少なくとも細胞外の生体内酸素濃度の測定には好適なプローブである。特に、完全に水素を重水素で置換したニトロキシドは、シグナル強度も大きく、好適なプローブであると判断した。 (2)マウス個体を対象として、生体内酸素濃度の測定を試みた。TEMPONE-D_<16>の溶液をガス透過性のプラスチックフィルムに封入したものをプローブとして、麻酔したマウスの皮下及び腹腔内に埋め込んだ。LバンドESR装置の共振器内の石英試料管にマウスを挿入し、ESR測定を行ったところ、十分な信号対雑音比で、シグナルを観測することができた。種々の濃度の酸素・窒素混合気を試料管に流して、グナルの線幅変化をピーク高さの変化から観測すると、酸素濃度に応答した変化を観測することができた。

Research Products

(27 results)

All Other

All Publications (27 results)

[Publications] T O'Hori,C-Y Wang,H Tokumaru,・・・,Y Kirino: "Action and binding of omega-conotoxin on the putative Ca channel of synaptosomal membrane from electric organ of Japanese ray" Neuroscience. 54. 1043-1050 (1993)
[Publications] T Tanaka,K Matsumoto,S Lee,・・・,Y Kirino: "Interaction of acidic amphiphilic α-helical peptide with lipid bilayers and their ion channel formation" Peptide Chemistry. 1992. 681-684 (1993)
[Publications] K Anzai,Y Yoshioka,Y Kirino: "Novel radioactive phospholipid probes as a tool for measurement of phospholipid translocation across biomembranes" Biochim.Biophys.Acta. 1151. 69-75 (1993)
[Publications] S Lee,T Iwata,H Oyagi,・・・,Y Kirino,G Sugihara: "Effect of salts on conformational change of basic amphipathic peptides and their channel-forming activity" Biochim.Biophys.Acta. 1151. 76-82 (1993)
[Publications] S Toda,N Hirashima,K Kuba,Y Kirino: "Spatio-temporal changes in intracellular Ca concentrations in giant neuronal cells from a terrestrial slug upon stimulation" Bioimages. 1. 95-103 (1993)
[Publications] K Anzai,M Masumi,K Kawasaki,Y Kirino: "Effects of lipid charge in planar bilayer membranes on the fusion of liposomes containg nystatin-ergosterol channels with the ・・・" J.Biochem.114. 487-491 (1993)
[Publications] H Tokumaru,C-Y Wang,N Hirashima,・・・,Y Kirino: "Action and binding of Ca channel blockers on the putative Ca channel of synptosomal plasma membrane from the electric ・・・" Annals New York Acad.Sci.707. 410-412 (1993)
[Publications] T Iwata,S Lee,O Oishi,・・・,Y Kirino,G Sugihara: "Design and synthesis of amphipathic 3_<10> helical peptides and their interactions with phospholipid bilayers and ion channel formation" J.Biol.Chem.(in press). (1994)
[Publications] S Lee,T Tanaka,K Anzai,Y Kirino,・・・,G Sugihara: "Two mode ion channels induced by interaction of acidic amphipathic α-helical peptides with lipid bilayers" Biochim.Biophys.Acta. (in press). (1994)
[Publications] K Shinozaki,K Anzai,Y Kirino,S Lee,H Aoyagi: "Ion channel activity of a synthetic peptide with a primary structure corresponding to the presumed pore-forming region of the ・・・" Biochem.Biophys.Res.Commun.(in press). (1994)
[Publications] K Anzai,C Takano,K Tanaka,Y Kirino: "Asymmetric lipid charge changes the substate conductance of the potassium channel from sarcoplasmic reticulum" Biochem.Biophys.Res.Commun.(in press). (1994)
[Publications] 安西和紀、桐野豊: "イオン輸送システム" 蛋白質核酸酵素・特集号「生体超分子システム」. 38. 1187-1192 (1993)
[Publications] Y Miura,H Utsumi,A Hamada: "Antioxidant activity of nitroxide radicals in lipid peroxidation" Arch.Biochem.Biophys.300. 148-156 (1993)
[Publications] K Takeshita,H Utsumi,A Hamada: "Whole mouse measurement of paramagnetism-loss of nitroxide free radicals in lung with L-band ESR spectrometer" Biochem.Mol.Biol.Int.29. 17-24 (1993)
[Publications] F Gomi,H Utsumi,A Hamada,M Matsuo: "Aging retards spin clearance from mouse brain and food restriction prevents its age-dependent retardation" Life Sicence. 52. 2027-2033 (1993)
[Publications] H Utsumi,K Takeshita,Y Miura,S Masuda,A Hamada: "In vivo ESR measurement of radical reaction in whole mouse.Influence of inspired oxygen and ischemia-reperfusion injury on ・・・" Free Rad.Res.Commun.19. s219-s225 (1993)
[Publications] H Utsumi,K Kiyoshige,S Shimbara,A Hamada: "Comparative studies on cytotoxicity of micropollutants in water.-Principle of cytotoxicity matrix-" Environ.Toxicol.Water Quality. (in press). (1994)
[Publications] 伊藤正吾、杉崎徹三、内海英雄、濱田昭: "生体計測用ESRを用いた腎組織内でのラジカル生成の検討" 医学のあゆみ. 164. 823-824 (1993)
[Publications] T Ozawa,J Ueda,A Hanaki: "Coper(II)-Albumin complex can activate hydrogen peroxide in the presence of biological reductants:First ESR evidence for ・・・" Molec.Biol.Int.29. 247-253 (1993)
[Publications] J Ueda,T Ozawa,M Miyazaki,Y Fujiwara: "SOD-like activity of complexes Ni(II) ion with some biologically important peptides and their novel reactions with hydrogen ・・・" Inorg.Chim.Acta. 214. 29-32 (1993)
[Publications] T Ozawa,J Ueda,Y Shimazu: "DNA single strand breakage by Cu(II) complexes and hydrogen per-oxide at physiological conditions" Biochem.Molec.Biol.Int.31. 455-461 (1993)
[Publications] J Ueda,T Ozawa,M Miyazaki,Y Fujiwara: "Activation of hydrogen peroxide by Cu(II) complexes with some histidine-containing peptides and their SOD-like activities" J.Inorg.Biochem.(in press). (1994)
[Publications] 小沢俊彦、湯川修身、花木昭、松島美一、小野寺加代子、・・: "銅(II)錯体による過酸化水素の活性化" 磁気共鳴と医学. 4. 29-32 (1993)
[Publications] N Shimamoto,K Kubo,T Watanabe,N Suzuki,・・・: "Pharmacologic profile of endothelin A/B antagonist,[Thr^<18>,γ-methylLeu^<19>]endothelin-1" J.Cardiovasc.Pharmacol.22. s107-s110 (1993)
[Publications] T Kikuchi,K Kubo,・・・,N Shimamoto,・・・,M Fujino: "Endothelin-1 analogues substituted at both position 18 and 19:Highly potent endothelin antagonists with no selectivity for ・・・" J.Med.Chem.36. 4087-4093 (1993)
[Publications] 桐野豊、他: "イオンチャネル1" メジカルビュー社, 230 (1993)
[Publications] 桐野豊、他: "レセプター" 朝倉書店, 944 (1993)

1993 Fiscal Year Annual Research Report

ESR法による細胞内酸素濃度測定法の開発

Principal Investigator

桐野 豊 東京大学, 薬学部, 教授 (10012668)

Research Products

[Publications] T O'Hori,C-Y Wang,H Tokumaru,・・・,Y Kirino: "Action and binding of omega-conotoxin on the putative Ca channel of synaptosomal membrane from electric organ of Japanese ray" Neuroscience. 54. 1043-1050 (1993)

[Publications] T Tanaka,K Matsumoto,S Lee,・・・,Y Kirino: "Interaction of acidic amphiphilic α-helical peptide with lipid bilayers and their ion channel formation" Peptide Chemistry. 1992. 681-684 (1993)

[Publications] K Anzai,Y Yoshioka,Y Kirino: "Novel radioactive phospholipid probes as a tool for measurement of phospholipid translocation across biomembranes" Biochim.Biophys.Acta. 1151. 69-75 (1993)

[Publications] S Lee,T Iwata,H Oyagi,・・・,Y Kirino,G Sugihara: "Effect of salts on conformational change of basic amphipathic peptides and their channel-forming activity" Biochim.Biophys.Acta. 1151. 76-82 (1993)

[Publications] S Toda,N Hirashima,K Kuba,Y Kirino: "Spatio-temporal changes in intracellular Ca concentrations in giant neuronal cells from a terrestrial slug upon stimulation" Bioimages. 1. 95-103 (1993)

[Publications] K Anzai,M Masumi,K Kawasaki,Y Kirino: "Effects of lipid charge in planar bilayer membranes on the fusion of liposomes containg nystatin-ergosterol channels with the ・・・" J.Biochem.114. 487-491 (1993)

[Publications] H Tokumaru,C-Y Wang,N Hirashima,・・・,Y Kirino: "Action and binding of Ca channel blockers on the putative Ca channel of synptosomal plasma membrane from the electric ・・・" Annals New York Acad.Sci.707. 410-412 (1993)

[Publications] T Iwata,S Lee,O Oishi,・・・,Y Kirino,G Sugihara: "Design and synthesis of amphipathic 3_<10> helical peptides and their interactions with phospholipid bilayers and ion channel formation" J.Biol.Chem.(in press). (1994)

[Publications] S Lee,T Tanaka,K Anzai,Y Kirino,・・・,G Sugihara: "Two mode ion channels induced by interaction of acidic amphipathic α-helical peptides with lipid bilayers" Biochim.Biophys.Acta. (in press). (1994)

[Publications] K Shinozaki,K Anzai,Y Kirino,S Lee,H Aoyagi: "Ion channel activity of a synthetic peptide with a primary structure corresponding to the presumed pore-forming region of the ・・・" Biochem.Biophys.Res.Commun.(in press). (1994)

[Publications] K Anzai,C Takano,K Tanaka,Y Kirino: "Asymmetric lipid charge changes the substate conductance of the potassium channel from sarcoplasmic reticulum" Biochem.Biophys.Res.Commun.(in press). (1994)

[Publications] 安西和紀、桐野豊: "イオン輸送システム" 蛋白質核酸酵素・特集号「生体 超分子システム」. 38. 1187-1192 (1993)

[Publications] Y Miura,H Utsumi,A Hamada: "Antioxidant activity of nitroxide radicals in lipid peroxidation" Arch.Biochem.Biophys.300. 148-156 (1993)

[Publications] K Takeshita,H Utsumi,A Hamada: "Whole mouse measurement of paramagnetism-loss of nitroxide free radicals in lung with L-band ESR spectrometer" Biochem.Mol.Biol.Int.29. 17-24 (1993)

[Publications] F Gomi,H Utsumi,A Hamada,M Matsuo: "Aging retards spin clearance from mouse brain and food restriction prevents its age-dependent retardation" Life Sicence. 52. 2027-2033 (1993)

[Publications] H Utsumi,K Takeshita,Y Miura,S Masuda,A Hamada: "In vivo ESR measurement of radical reaction in whole mouse.Influence of inspired oxygen and ischemia-reperfusion injury on ・・・" Free Rad.Res.Commun.19. s219-s225 (1993)

[Publications] H Utsumi,K Kiyoshige,S Shimbara,A Hamada: "Comparative studies on cytotoxicity of micropollutants in water.-Principle of cytotoxicity matrix-" Environ.Toxicol.Water Quality. (in press). (1994)

[Publications] 伊藤正吾、杉崎徹三、内海英雄、濱田昭: "生体計測用ESRを用いた腎組織内でのラジカル生成の検討" 医学のあゆみ. 164. 823-824 (1993)

[Publications] T Ozawa,J Ueda,A Hanaki: "Coper(II)-Albumin complex can activate hydrogen peroxide in the presence of biological reductants:First ESR evidence for ・・・" Molec.Biol.Int.29. 247-253 (1993)

[Publications] J Ueda,T Ozawa,M Miyazaki,Y Fujiwara: "SOD-like activity of complexes Ni(II) ion with some biologically important peptides and their novel reactions with hydrogen ・・・" Inorg.Chim.Acta. 214. 29-32 (1993)

[Publications] T Ozawa,J Ueda,Y Shimazu: "DNA single strand breakage by Cu(II) complexes and hydrogen per-oxide at physiological conditions" Biochem.Molec.Biol.Int.31. 455-461 (1993)

[Publications] J Ueda,T Ozawa,M Miyazaki,Y Fujiwara: "Activation of hydrogen peroxide by Cu(II) complexes with some histidine-containing peptides and their SOD-like activities" J.Inorg.Biochem.(in press). (1994)

[Publications] 小沢俊彦、湯川修身、花木昭、松島美一、小野寺加代子、・・: "銅(II)錯体による過酸化水素の活性化" 磁気共鳴と医学. 4. 29-32 (1993)

[Publications] N Shimamoto,K Kubo,T Watanabe,N Suzuki,・・・: "Pharmacologic profile of endothelin A/B antagonist,[Thr^<18>,γ-methylLeu^<19>]endothelin-1" J.Cardiovasc.Pharmacol.22. s107-s110 (1993)

[Publications] T Kikuchi,K Kubo,・・・,N Shimamoto,・・・,M Fujino: "Endothelin-1 analogues substituted at both position 18 and 19:Highly potent endothelin antagonists with no selectivity for ・・・" J.Med.Chem.36. 4087-4093 (1993)

[Publications] 桐野豊、他: "イオンチャネル1" メジカルビュー社, 230 (1993)

[Publications] 桐野豊、他: "レセプター" 朝倉書店, 944 (1993)

桐野豊東京大学, 薬学部, 教授 (10012668)

[Publications] 安西和紀、桐野豊: "イオン輸送システム" 蛋白質核酸酵素・特集号「生体超分子システム」. 38. 1187-1192 (1993)