イソキノリンによるパーキンソン病モデルの作成

1996 Fiscal Year Annual Research Report

• Project/Area Number: 08256213
• Research Institution: Nagoya Institute of Technology
• Principal Investigator: 直井 信 名古屋工業大学, 工学部, 教授 (50022786)
• Keywords: パーキンソン病 / アポトーシス / 内在性神経毒 / N-メチルサルソリノール / ドーパミン / 細胞死 / 黒質線状体 / (R)サルソリノール合成酵素

Research Abstract

パーキンソン病(PD)は黒質線状体のドーパミン細胞の選択的な細胞死により惹起される疾病であり、細胞死の原因としてはドーパミンから生成される1(R),2(N)-dimethyl-DHTIQ[N-methyl(R)salsolinol,NM(R)Sal]のみがラットにPD症状を惹起することを報告した。この物質のラット脳内持続投与により、黒質ドーパミンニューロンが選択的に脱落した。またPDの患者の脳脊髄液の分析によりNM(R)Salが有意に増加していることを発見した。
ドーパミン細胞モデルである培養細胞SH-SY5Yを用い神経毒NM(R)Salによる細胞死の機序を検討したところ、NMSalのみがDNAの切断を惹起した。(R)型が(S)型より高度に傷害をおこし、タンパク合成を阻害することで損傷を防止できたことから、アポトーシスの誘導によることが明かとなった。このアポトーシスは抗酸化剤と抗酸化酵素により防止できたことから、酸化的ストレスがアポトーシスを誘発していることが示唆された。
またヒト脳、脳脊髄液、脳室液には(R)型のSalとNMSalのみが存在し、(R)Salが脳全体に分布していたのに対し、NM(R)Salは黒質線状体にまたNM(R)Salの酸化生成物1,2-dimethyl-6,7-dihydroxyisoquinolinium ionは黒質に限局して蓄積していた。この事から、NM(R)Salを生合成する酵素の精製を試みた。ヒト脳からドーパミンとアセトアルデヒドから(R)型のSalを生成する(R)Salsolinol synthaseが精製された。さらにヒト脳に(R)Salに特異性をもつN-methyltransferaseが存在することを発見した。
これらの結果はNM(R)Salがヒト脳黒質線状体に内在する神経毒で長期の蓄積の後、ドーパミン細胞にアポトーシスを惹起しPDを発症させる可能性を示唆している。

Research Products

• [Publications] 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-dihydroxyisoquinolinium ion. accumulate in the nigro-striatal system of the brain" Neurosciences Letters in press. (1997)
• [Publications] Wakako Maruyama: "An endogenous dopaminergic neuritixn, N-methyl (R) salsolinol, induces DNA damage in human dopaminergic neuroblastoma SH-SY5Y" Journal of Neurochemisty in press23GE02:1997.
• [Publications] 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)
• [Publications] 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)
• [Publications] Philippe Dostert: "Effects of dopamine-derived isoquinolines on hydroxy radical formation by dopamine autoxidation" Biogenic Amines. 12. 149-160 (1996)
• [Publications] Makoto Naoi: "Oxidation of N-methyl (R) salsolinol : Involvement to neurotoxicity abd neuroprotection by endogenous catechol isoquinolines." Journal of Neural Transmission. (1996)
• [Publications] Wakakot Maruyama: "Apoptosis of dopamine cells induced by N-methyl (R) salsolinol" Journal of Neural Transmission. (1996)
• [Publications] Wakako Maruyama: "A dopaminergic neurotoxin, (R)-N--methylsalsolinol increases in parkiosnian CSF" Annalus of Neurology. 40. 119-122 (1996)
• [Publications] Makoto Naoi: "A novel enzyme enatio-selectively synthesizes (R) salsolinol, a precursor of a dopaminergic neurotoxin, N-methyl (R) salsolinol" Neuroscience Letters. 212. 183-186 (1996)
• [Publications] Kazuo Matsubara: "Modulation of tryptophan hydroxylase activity in vitro by ethanol depends on biopterin cofactor concentration" Alcohol. 13. 455-459 (1996)
• [Publications] Makoto Naoi: "Dopamine-derived endogenous 1(R), 2(N)-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, N-methyl-(R)-salsolinol, induces parkinsonism in rat ; Biochemical, pathological and behavioral studies" Brain Research. 709. 285-295 (1996)
• [Publications] 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)
• [Publications] Wakako Maruyama: "The mechanism of L-DOPA and L-threo-dihydroxyphenylserine 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)
• [Publications] Hideki Miura: "Efects of moclobemide on forced-swimming stress and brain monoamine levels in mice." Pharmacology Biochemisty and Behavior. 53. 469-475 (1996)
• [Publications] Yulin Deng: "Assay for the (R)-and (S)-enenatiomers of salsolinols in biological samples and foods with ion-pair HPLC using b-cyclodextrins as a chiral phase additive" Journal of Chromatography B. (1996)
• [Publications] Wakako Maruyama: "Stereospecific occurrence of a parkinssonim-inducing catechol isoquinoline, N-methyl (R)-salsolinol in the human intraventricular fluid" Journal of Neural Transmission [GenSect]. 103. 1069-1076 (1996)
• [Publications] Yulin Deng: "Mechanism of Enantioseperation ofSalsolinols, Endogenous Neurotoxins in Human Brain, with Ion-Pair Chromatography Using b-cyclodextrin as a Mobile Phase Additive" Analytical Chemistry. 68. 2826-2831 (1996)
• [Publications] Tetsuo Ohkuwa: "Glutathione status and reactive oxygen gencration in tissue of young and old exercised rats" Acta Physiologcal Scandinavica. (1996)
• [Publications] 丸山和佳子: "Parkinson病" 最新医学. 51. 1278-1287 (1996)
• [Publications] 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)
• [Publications] Wakako Maruyama: "Non-enzymatic oxidation of 1,2(N)-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines into 1,2(N)-dimethyl-6,7-dihydroxyisoquinolinium ion, produces hydroxyl radicals" Free Radical Biologycal Medisne. 19. 67-75 (1995)
• [Publications] Tetsuo Ohkuwa: "Hydroxyl radical formation in diabetic rats induced by streptozotocin." Life Sciences. 56. 1789-1798 (1995)
• [Publications] Mitsuharu Kajita: "Detection of 1-phenyl-N-methyl-1,2,3,4-tetrahydroisoquinoline and 1-phenyl-1,2,3,4-tetrahydroisoquinoline in parkinsonian human brain by gas chromatography/tandem mass spectromet" Journal of Chromatography B. 669. 345-351 (1995)
• [Publications] 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)
• [Publications] Shinnichi Hasegawa: "Inhibition fo type A monoamine oxidase by N-methyl b-carbolinium ion" Biogenic Amines. 11. 295-303 (1995)
• [Publications] Makoto Naoi: "Enzymanic oxidation of dopaminergic neurotoxin, 1(R), 2 (N)-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline into 1,2 (N)-dimethyl-6,7-dihydroxyisoquinolinium ion" Life Sciences. 57. 1061-1066 (1995)
• [Publications] Shinya Kitagawa: "Manipulation of a single cell with microcapillary tubing based on its electophoretic mobility" Electrophoresis. 16. 1364-1268 (1995)
• [Publications] Masayuki Shiratori: "Application of liposome immune lysis assay for studies on anti-ganglioside antibodies." Journal of Clinical Biochemistry and Nutrition. 19. 45-54 (1995)
• [Publications] 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)
• [Publications] Wakako Maruyama: "Dopamine-derived 1-methyl-6,7-dihydroxyisoquinolines as hydroxyl radical promoters and scavengers in the rat brain : in vivo and in vitro studies" Jounal of Neurochemstry. 64. 2635-2643 (1996)
• [Publications] Makoto Naoi: "Pharmacology of endogenous nerurotoxins" An animal model of parkinsosn′s prepared by endogenous N-methyl (R) salsolinol, (1997)
• [Publications] Makoto Naoi: "Pharmacology of endogenous nerurotoxins" An animal model of parkinsosn′s disease prepared by endogenous N-methyl (R) salsolinol, (1997)
• [Publications] Makoto Naoi: "Molecular Toxiology" Cytotoxicity of an endogenous dopaminergic neurotixn, N-methyl (R) salsolinol : relevance to parkineson′s disease, (1997)
• [Publications] Wakako Maruyama: "Molecular Toxiology" Quantitative analysis of apoptotic neuronal cell death by single cell gel electrophoresis assay, (1997)
• [Publications] 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)
• [Publications] 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)
• [Publications] Wakako Maruyama: "Progress in HPLC, Volume 4, ″Multielectrode EC Array Applications to Trace Analysis of Neurotransittes. Pharmaceuticals and Environmental Markes″" Application of Multi-Electrode Detection System to Monitoring of Drug Therapy for Neurological Diseases, (1996)
• [Publications] 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)
• [Publications] 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)
• [Publications] Wakako Maruyama: "Alzheimer′s and Parkinson′s Diseases : Recent Developments" Dopamine-derived isoquinolines asdopaminergic neurotoxins and oxidative stress, 10 (1996)
• [Publications] K.Suzuki: "Alzheimer′s and Parkinson′s Diseases : Recent Developments" Inhibitory effects of endogenous tetrahydroisoquinolines on mitochondrial respiration in mouse brain., 13 (1996)
• [Publications] Makoto Naoi: "Progress in Brain Research" Dopamine-derived 6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines ; Oxidation and Neurotoxicity, 13 (1995)
• [Publications] 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)