1996 年度実績報告書

活性酸素peroxynitriteによる神経細胞死の機序と予防の研究

研究課題

研究課題/領域番号	08838010
研究機関	名古屋工業大学
研究代表者	直井信名古屋工業大学, 工学部, 教授 (50022786)
キーワード	活性酸素 / 神経細胞死 / 神経変成疾患 / nitric oxide / peroxinitrite / apoptosis / DNA損傷 / 抗酸化剤
研究概要	痴呆や運動障害を惹起する神経変性疾患および血管性痴呆などの疾患における神経細胞の傷害が、活性酸素により惹起されるとの説が有力となっている。活性酸素の中でも細胞毒性の高い水酸化ラジカルとperoxynitriteが重要な役割を担っているとされる。superoxideとnitric oxide(NO)から生成されるperoxynitriteは最近注目されている活性酸素であり、peroxinitriteのL-tyrosineとの反応生成物、3-nitro-L-tyrosineの定量法を開発し、ヒト脳での存在を報告した。3-nitro-L-tyrosineはperoxynitriteによる細胞傷害のマーカーであり、疾患との関連を検討した。今回神経細胞に対するperoxynitriteの直接作用を明らかにするため、ドーパミン細胞のモデルである培養細胞SH-SY5Y細胞を対する細胞毒性を検討した。このためDNAの損傷を定量するsingle cell gelelectrophoresis(Comet法)と、細胞内のATP産生を定量するAlamar Blue法を用い検討した。 NOの発生剤である3-{{±)-(E)-ethyl-2'-[(E)-hydroxyimino]-5-nitro-3-hexenecarbamonyl}-pyridine(NOR-4)と、peroxinitriteの発生剤N-morpholono sydnonimine(SIN-1)を用い細胞毒性を検討したところ、濃度依存的にDNAの損傷を惹起した。このDNA損傷はタンパクの生成を介するアポトーシスであった。このアポトーシスを予防できる薬剤を発見し、現在その構造活性相関と予防の機序を検討している。これらの結果は神経変成疾患において、NOからperoxynitriteへの生合成により活性酸素がアポトーシスを惹起し、神経細胞の死をもたらす可能性を示唆している。

研究成果
(42件)

すべてその他

すべて文献書誌 (42件)

[文献書誌] Wakako Maruyama: "A dopaminergic neurotoxin,1 (R),2 (N) -dimethyl-6,7-dihydro-1,2,3,4-tetrahydroisoquinoline,N-methyl (R) salsolinol,and its oxidation product,1,2 (N) -dimethyl-6,7-dihydroxysoquinolinium ion. accumulate in the nigro-striatal system of the brain" Neurosciences Letters in press. (1997)
[文献書誌] Wakako Maruyama: "An endogenous dopaminergic neuritixn,N-methyl (R) salsolinol,induces DNA damage in human dopaminergic neuroblastoma SH-SY5Y" Journal of Neurochemistry in press. (1997)
[文献書誌] Makoto Naoi: "N-methyl- (R) salsolinol as a dopaminergic neurotoxin : from an animal model to an early marker of parkison's disease" Journal of Neural Transmission,Supplement. 50. 89-105 (1997)
[文献書誌] Makoto Naoi: "Animal model of Parkinson's disease induced by naturally-occurring 1 (R),2 (N) -dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline" Biogenic Amines. 12. 135-147 (1996)
[文献書誌] Philippe Dostert: "Effects of dopamine-derived isoquinolines on hydroxy radical formation by dopamine autoxidation" Biogenic Amines. 12. 149-160 (1996)
[文献書誌] Makoto Naoi: "Oxidation of N-methyl (R) salsolinol : Involvement to neurotoxicity abd neuroprotection by endogenous catechol isoquinolines" Journal of Neural Transmission. (1996)
[文献書誌] Wakako Maruyama: "Apoptosis of dopamine cells induced by N-methyl (R) salsolinol" Journal of Neural Transmission. (1996)
[文献書誌] Wakako Maruyama: "A dopaminergic neurotoxin,(R) -N-methylsalsolinol increases in parkiosnian CSF" Annalus of Neurology. 40. 119-122 (1996)
[文献書誌] Makoto Naoi: "A novel enzyme enation-selectively synthesizes (R) salsolinol,a precursor of a dopaminergic neurotoxin,N-methyl (R) salsolinol" Neuroscience Letters. 212. 183-186 (1996)
[文献書誌] Kazuo Matsubara: "Modulation of tryptophan hydroxylase activity in vitro by ethanol depends on biopterin cofact or concentration" Alcohol. 13. 455-459 (1996)
[文献書誌] Makoto Naoi: "Dopamine-derived endogenous 1 (R),2 (N) -dimethyl-6,7-dihy droxy-1,2,3,4-tetrahy droisoquinoline,N-methyl- (R) -salsolinol,induces parkinsonism in rat ; Biochemical,pathological and behavioral studies" Brain Research. 709. 285-295 (1996)
[文献書誌] Wakako Maruyama: "Identification of 3-nitro-L-tyrosine,a product of nitric oxide and superoxide,as an indicator of oxidative stress in the human brain" Journal of Chromatography Biomedical Application. 626. 153-158 (1996)
[文献書誌] Wakako Maruyama: "The mechanism of L-DOPA and L-threo-dihy droxyphenylserine and their effects on monoamine in the human brain : Analysis of the intraventricular fluid of parkinsonian patients" Journal of Neurological Sciences. 139. 141-148 (1996)
[文献書誌] Hideki Miura: "Effects of moclobemide on forced-swimming stress and brain monoamine levels in mice." Pharmacology Biochemisty and Behavior. 53. 469-475 (1996)
[文献書誌] Yulin Deng: "Assay of the (R) -and (S) -enenatiomers of salsolinols in biological samples and foods with ion-pair HPLC using b-cyclodextrins as a chiral pahse additive" Journal of Chromatography B. (1996)
[文献書誌] Wakako Maruyama: "Stereospecific occurrence of a parkinsonism-inducing catechol isoquinoline,N-methyl (R) -salsolinol in the human intraventricular fluid" Journal of Neural Transmission [GenSect]. 103. 1069-1076 (1996)
[文献書誌] Yulin Deng: "Mechanism of Enantioseperation of Salsolinols,Endogenous Neurotoxins in Human Brain,with Ion-Pair Chromatography Using b-cyclodextrin as a Mobile Phase Additive" Analytical Chemictry. 68. 2826-2831 (1996)
[文献書誌] Tetsuo Ohkuwa: "Glutathione status and reactive oxygen generation in tissue of young and old exercised rats" Acta Physiological Scandinavica. (1996)
[文献書誌] 丸山和佳子: "Parkinson病" 最新医学. 51. 1278-1287 (1996)
[文献書誌] 直井信: "パーキンソン病の今日の課題。「病因論」、1。神経毒" Progress in Medicine. 16. 2740-2745 (1996)
[文献書誌] Wakao Maruyama: "Non-enzymatic oxidation of 1,2 (N) -dimethyl-6,7-dihydroxy-1,2,3,4-tetrahy droisoquinolines into 1,2 (N) -dimethyl-6,7-dihy droxyisoquinolinium ion,produces hydroxy radicals" Free Radical Biologycal Medisne. 19. 67-75 (1995)
[文献書誌] Tetsuo Ohkuwa: "Hydroxyl radical formation in diabetic rats induced by streptozotocin." Life Sciences. 56. 1789-1798 (1995)
[文献書誌] Mitsuharu Kajita: "Detection of 1-phenyl-N-methyl-1,2,3,4-tetrahy droisoquinoline and 1-phenyl-1,2,3,4-tetrahy droisoquinoline in parkinsonian human brain by gas chromatography/tandem mass spectrom" Journal of Chromatography B. 669. 345-351 (1995)
[文献書誌] Yulin Deng: "Determination of (R) -and (S) -enantiomers of salsolinol and N-methylsalsolinol by use of a chiral HPLC column" Journal of Chromatography B. 670. 47-54 (1995)
[文献書誌] Shinnichi Hasegawa: "Inhibition of type A monoamine oxidase by N-methyl b-carbolinium ion" Biogenic Amines. 11. 295-303 (1995)
[文献書誌] Makoto Naoi: "Enzymatic oxidation of dopaminergic neurotoxin,1 (R),2 (N) -dimethyl-6,7-dihy droxy-1,2,3,4-tetrahy droisoquinoline into 1,2 (N) -dimethyl-6,7-dihy droxyisoquinolinium ion" Life Sciences. 57. 1061-1066 (1995)
[文献書誌] Shinya Kitagawa: "Manipulation of a single cell with microcapillary tubing based on its electophoretic mobility." Electrophoresis. 16. 1364-1268 (1995)
[文献書誌] Masayuki Shiratori: "Application of liposome immune lysis assay for studies on anti-ganglioside antibodies." Journal of Clinical Biochemistry and Nutrition. 19. 45-54 (1995)
[文献書誌] Kazuo Matsubara: "N-Methyl-4-phenylpyridinium and an endogenously formed analog,N-methylated bcarbolinium,inhibit striatal tyrosine hydroxylation in freely moving rats." Neurosciences Letters. 199. 199-202 (1995)
[文献書誌] Wakako Maruyama: "Dopamine-derived 1-methyl-6,7-dihy droxyisoquinolines as hydroxyl radical promoters and scavengers in the rat brain : in vivo and in vitro studies" Jounal of Neurochemstry. 64. 2635-2643 (1995)
[文献書誌] Makoto Naoi: "Pharmacology of endogenous nerurotoxins" An animal model of parkinson's disease prepared by endogenous N-methyl (R) salsolinol, (1997)
[文献書誌] Makoto Naoi: "Molecular Toxiology" Cytotoxicity of an endogenous dopaminergic neurotixn,N-methyl (R) salsolinol : relevance to parkineson's disease, (1997)
[文献書誌] Wakako Maruyama: "Molecular Toxiology" Quantitative analysis of apoptotic neuronal cell death by single cell gel electriphoresis assay, (1997)
[文献書誌] Wakako Maruyama: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Analysis of Enatiomers of Endogenous Neurotoxins in Clinical Samples by Multi-Electrode Detection System With a Chiral Column, (1996)
[文献書誌] Makoto Naoi: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Application of Multi-Electrode Detection System to Neurochemistry ; In Vivo Microdialysis Study, (1996)
[文献書誌] Wakako Maruyama: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Application of Multi-Electrode Detection System to Monitoring of Drug Therapy for Neurological Diseases, (1996)
[文献書誌] Yulin Deng: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Enatioseparation of Biologically Active Compounds by HPLC, (1996)
[文献書誌] Makoto Naoi: "Alzheimer's and Parkinson's Diseases : Recent Developments" Metabolic bioactivation of endogenous isoquinolines as dopaminergic neurotoxins to elicit Parkinson's Disease., 7 (1996)
[文献書誌] Wakako Maruyama: "Alzheimer's and Parkinson's Diseases : Recent Developments" Dopamine-derived isoquinolines as dopaminergic neurotoxins and oxidative stress, 10 (1996)
[文献書誌] K.Suzuki: "Alzheimer's and Parkinson's Diseases : Recent Developments" Inhibitory effects of endogenous tetrahy droisoquinolines on mitochondrial respiration in mouse brain., 13 (1996)
[文献書誌] Makoto Naoi: "Progress in Brain Research" Dopamine-derived 6,7-dihy droxy-1,2,3,4-tetrahydroisoquinolines ; Oxidation and Neurotoxicity, 13 (1995)
[文献書誌] 直井信: "神経細胞死Up to Date基礎と臨床からのアプローチ" 神経毒による神経細胞死の機構, 20 (1995)

1996 年度 実績報告書

活性酸素peroxynitriteによる神経細胞死の機序と予防の研究

研究代表者

直井 信 名古屋工業大学, 工学部, 教授 (50022786)

研究成果

[文献書誌] Wakako Maruyama: "An endogenous dopaminergic neuritixn,N-methyl (R) salsolinol,induces DNA damage in human dopaminergic neuroblastoma SH-SY5Y" Journal of Neurochemistry in press. (1997)

[文献書誌] Makoto Naoi: "N-methyl- (R) salsolinol as a dopaminergic neurotoxin : from an animal model to an early marker of parkison's disease" Journal of Neural Transmission,Supplement. 50. 89-105 (1997)

[文献書誌] Makoto Naoi: "Animal model of Parkinson's disease induced by naturally-occurring 1 (R),2 (N) -dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline" Biogenic Amines. 12. 135-147 (1996)

[文献書誌] Philippe Dostert: "Effects of dopamine-derived isoquinolines on hydroxy radical formation by dopamine autoxidation" Biogenic Amines. 12. 149-160 (1996)

[文献書誌] Makoto Naoi: "Oxidation of N-methyl (R) salsolinol : Involvement to neurotoxicity abd neuroprotection by endogenous catechol isoquinolines" Journal of Neural Transmission. (1996)

[文献書誌] Wakako Maruyama: "Apoptosis of dopamine cells induced by N-methyl (R) salsolinol" Journal of Neural Transmission. (1996)

[文献書誌] Wakako Maruyama: "A dopaminergic neurotoxin,(R) -N-methylsalsolinol increases in parkiosnian CSF" Annalus of Neurology. 40. 119-122 (1996)

[文献書誌] Makoto Naoi: "A novel enzyme enation-selectively synthesizes (R) salsolinol,a precursor of a dopaminergic neurotoxin,N-methyl (R) salsolinol" Neuroscience Letters. 212. 183-186 (1996)

[文献書誌] Kazuo Matsubara: "Modulation of tryptophan hydroxylase activity in vitro by ethanol depends on biopterin cofact or concentration" Alcohol. 13. 455-459 (1996)

[文献書誌] Makoto Naoi: "Dopamine-derived endogenous 1 (R),2 (N) -dimethyl-6,7-dihy droxy-1,2,3,4-tetrahy droisoquinoline,N-methyl- (R) -salsolinol,induces parkinsonism in rat ; Biochemical,pathological and behavioral studies" Brain Research. 709. 285-295 (1996)

[文献書誌] Wakako Maruyama: "Identification of 3-nitro-L-tyrosine,a product of nitric oxide and superoxide,as an indicator of oxidative stress in the human brain" Journal of Chromatography Biomedical Application. 626. 153-158 (1996)

[文献書誌] Wakako Maruyama: "The mechanism of L-DOPA and L-threo-dihy droxyphenylserine and their effects on monoamine in the human brain : Analysis of the intraventricular fluid of parkinsonian patients" Journal of Neurological Sciences. 139. 141-148 (1996)

[文献書誌] Hideki Miura: "Effects of moclobemide on forced-swimming stress and brain monoamine levels in mice." Pharmacology Biochemisty and Behavior. 53. 469-475 (1996)

[文献書誌] Yulin Deng: "Assay of the (R) -and (S) -enenatiomers of salsolinols in biological samples and foods with ion-pair HPLC using b-cyclodextrins as a chiral pahse additive" Journal of Chromatography B. (1996)

[文献書誌] Wakako Maruyama: "Stereospecific occurrence of a parkinsonism-inducing catechol isoquinoline,N-methyl (R) -salsolinol in the human intraventricular fluid" Journal of Neural Transmission [GenSect]. 103. 1069-1076 (1996)

[文献書誌] Yulin Deng: "Mechanism of Enantioseperation of Salsolinols,Endogenous Neurotoxins in Human Brain,with Ion-Pair Chromatography Using b-cyclodextrin as a Mobile Phase Additive" Analytical Chemictry. 68. 2826-2831 (1996)

[文献書誌] Tetsuo Ohkuwa: "Glutathione status and reactive oxygen generation in tissue of young and old exercised rats" Acta Physiological Scandinavica. (1996)

[文献書誌] 丸山和佳子: "Parkinson病" 最新医学. 51. 1278-1287 (1996)

[文献書誌] 直井信: "パーキンソン病の今日の課題。「病因論」、1。神経毒" Progress in Medicine. 16. 2740-2745 (1996)

[文献書誌] Wakao Maruyama: "Non-enzymatic oxidation of 1,2 (N) -dimethyl-6,7-dihydroxy-1,2,3,4-tetrahy droisoquinolines into 1,2 (N) -dimethyl-6,7-dihy droxyisoquinolinium ion,produces hydroxy radicals" Free Radical Biologycal Medisne. 19. 67-75 (1995)

[文献書誌] Tetsuo Ohkuwa: "Hydroxyl radical formation in diabetic rats induced by streptozotocin." Life Sciences. 56. 1789-1798 (1995)

[文献書誌] Mitsuharu Kajita: "Detection of 1-phenyl-N-methyl-1,2,3,4-tetrahy droisoquinoline and 1-phenyl-1,2,3,4-tetrahy droisoquinoline in parkinsonian human brain by gas chromatography/tandem mass spectrom" Journal of Chromatography B. 669. 345-351 (1995)

[文献書誌] Yulin Deng: "Determination of (R) -and (S) -enantiomers of salsolinol and N-methylsalsolinol by use of a chiral HPLC column" Journal of Chromatography B. 670. 47-54 (1995)

[文献書誌] Shinnichi Hasegawa: "Inhibition of type A monoamine oxidase by N-methyl b-carbolinium ion" Biogenic Amines. 11. 295-303 (1995)

[文献書誌] Makoto Naoi: "Enzymatic oxidation of dopaminergic neurotoxin,1 (R),2 (N) -dimethyl-6,7-dihy droxy-1,2,3,4-tetrahy droisoquinoline into 1,2 (N) -dimethyl-6,7-dihy droxyisoquinolinium ion" Life Sciences. 57. 1061-1066 (1995)

[文献書誌] Shinya Kitagawa: "Manipulation of a single cell with microcapillary tubing based on its electophoretic mobility." Electrophoresis. 16. 1364-1268 (1995)

[文献書誌] Masayuki Shiratori: "Application of liposome immune lysis assay for studies on anti-ganglioside antibodies." Journal of Clinical Biochemistry and Nutrition. 19. 45-54 (1995)

[文献書誌] Kazuo Matsubara: "N-Methyl-4-phenylpyridinium and an endogenously formed analog,N-methylated bcarbolinium,inhibit striatal tyrosine hydroxylation in freely moving rats." Neurosciences Letters. 199. 199-202 (1995)

[文献書誌] Wakako Maruyama: "Dopamine-derived 1-methyl-6,7-dihy droxyisoquinolines as hydroxyl radical promoters and scavengers in the rat brain : in vivo and in vitro studies" Jounal of Neurochemstry. 64. 2635-2643 (1995)

[文献書誌] Makoto Naoi: "Pharmacology of endogenous nerurotoxins" An animal model of parkinson's disease prepared by endogenous N-methyl (R) salsolinol, (1997)

[文献書誌] Makoto Naoi: "Molecular Toxiology" Cytotoxicity of an endogenous dopaminergic neurotixn,N-methyl (R) salsolinol : relevance to parkineson's disease, (1997)

[文献書誌] Wakako Maruyama: "Molecular Toxiology" Quantitative analysis of apoptotic neuronal cell death by single cell gel electriphoresis assay, (1997)

[文献書誌] Makoto Naoi: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Application of Multi-Electrode Detection System to Neurochemistry ; In Vivo Microdialysis Study, (1996)

[文献書誌] Wakako Maruyama: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Application of Multi-Electrode Detection System to Monitoring of Drug Therapy for Neurological Diseases, (1996)

[文献書誌] Yulin Deng: "Progress in HPLC,Volume 4,"Multielectrode EC Array Applications to Trace Analysis of Neurotransmitters,Pharmaceuticals and Environmental Markers"" Enatioseparation of Biologically Active Compounds by HPLC, (1996)

[文献書誌] Makoto Naoi: "Alzheimer's and Parkinson's Diseases : Recent Developments" Metabolic bioactivation of endogenous isoquinolines as dopaminergic neurotoxins to elicit Parkinson's Disease., 7 (1996)

[文献書誌] Wakako Maruyama: "Alzheimer's and Parkinson's Diseases : Recent Developments" Dopamine-derived isoquinolines as dopaminergic neurotoxins and oxidative stress, 10 (1996)

[文献書誌] K.Suzuki: "Alzheimer's and Parkinson's Diseases : Recent Developments" Inhibitory effects of endogenous tetrahy droisoquinolines on mitochondrial respiration in mouse brain., 13 (1996)

[文献書誌] Makoto Naoi: "Progress in Brain Research" Dopamine-derived 6,7-dihy droxy-1,2,3,4-tetrahydroisoquinolines ; Oxidation and Neurotoxicity, 13 (1995)

[文献書誌] 直井信: "神経細胞死Up to Date基礎と臨床からのアプローチ" 神経毒による神経細胞死の機構, 20 (1995)

1996 年度実績報告書

直井信名古屋工業大学, 工学部, 教授 (50022786)