1996 Fiscal Year Annual Research Report

ヒト脳由来の新規神経保護活性物質の探索

Research Project

Project/Area Number	08877104
Research Institution	Kyoto University
Principal Investigator	下濱俊京都大学, 医学研究科, 助手 (60235687)
Keywords	アルツハイマー病 / 神経変性 / 細胞死 / 神経保護活性物質 / 初代培養中脳神経細胞 / ドパミンニューロン / 脳抽出物 / 脳由来神経栄養因子
Research Abstract	アルツハイマー病(AD)、パーキンソン病、脊髄小脳変性症、筋萎縮性側索硬化症などの神経難病に共通の解決すべき最大の問題は、神経細胞が変性し、神経細胞死に至る機序の解明とともに、この神経細胞の変性を防止し、神経細胞死を予防する方法の開拓である。本研究では、in vitroの神経細胞死モデルをスクリーニング系として用い、ヒト脳より新規の神経保護活性物質の単離を試みた。ラット初代培養中脳神経細胞を用いたスクリーニングバイオアッセイ系を確立した。培養細胞を定法により4日間維持し、その後4日間血清不含培地にヒト脳抽出物を添加した。細胞固定後ドパミン(DA)ニューロン、非DAニューロンの生存率を検定した。 DAニューロン:正常ヒト脳由来試料添加群では1-3μg protein/mlの低濃度投与で生存率が低下し、10μg protein/ml以上の高濃度では生存率はほとんど低下しなかった。一方、AD脳由来試料添加群では、低濃度側で生存率が若干増加する傾向を示し、高濃度になるにつれ生存率が減少する傾向を示した。3μg protein/mlの濃度でAD脳由来試料添加群の生存率が正常ヒト脳由来試料添加群と比較して有意な高値を示した。非DAニューロン:正常ヒト脳由来試料添加群、AD脳由来試料添加群ともに1μg protein/mlの低濃度投与で生存率が低下し、濃度をあげると生存率が回復し、さらに濃度をあげると著しい生存率の低下が認められた。AD脳由来試料添加群では30μg protein/mlから生存率が低下し、正常ヒト脳由来試料添加群と有意な差が認められた。以上の結果から、培養中脳細胞において、AD脳由来抽出物中にDAニューロンに対して生存促進作用のある物質が存在する可能性が示唆された。今後、この物質を同定し、既知の脳由来神経栄養因子とは異なる新たな神経保護活性物質であるか否かの検討を要する

Research Products
(14 results)

All Other

All Publications (14 results)

[Publications] Sawada H: "Methyl phenylpyoldium ion(MPP^+)enbances glutamate-induced cytotoxicity against dopaminergic neurons in cultuned rat mesen cephalon" Journal of Neuroscience Research. 43. 55-62 (1996)
[Publications] Sawada H: "Different mechanisms of glutamate-induced neuronal death between dopaminergic and non-dopaminergic neurons in rat mesencephalic culture" Journal of Neuroscience Research. 43. 503-510 (1996)
[Publications] Shimohama S: "Nicotine-induced protection against glutamate cytotoxicity:Nicotinic cholinergic receptor・mediated inhibition of nitric oxide formation" Annals of New York Academy of Sciences. 777. 356-361 (1996)
[Publications] Shimohama S: "Changers of neurocaloin,a calcium-binding protein,in the brain of patients with Alzheimer's disease" Brain Research. 716. 233-236 (1996)
[Publications] Matsushima H: "Ca^<2+>-dependent and-independent protein kinase C changes in the brain of patients with Alzheimer's disease" Journal of Neurochemistry. 67. 317-323 (1996)
[Publications] Chachin M: "Assessment of protein kinase C mRNA levels in Alzheimer's disease brains" Japanese Journal of Pharmacology. 71. 175-177 (1996)
[Publications] Fujimoto S: "Zinc-ion-dependent acid phosphatase exhibits maguesium-ion-dependent myo-inositol-1-phosphatase activity" Biol.Pharm.Bull. 19. 882-885 (1996)
[Publications] Kawakami N: "Tyrosine phosphorylation and translocation of phospholipase C-γ_2 in polyrmorphonuclear leukocytes treated with pervanadate" Biochim Biophys Acta. 1314. 167-174 (1996)
[Publications] Sawada H: "Mechanism of resistance to NO-induced neurotoxicity in cultured rat dopaminergic neurons" Journal of Neuroscience Research. 46. 509-518 (1996)
[Publications] Kihara T: "Nicotinic receptor stimulation protects neurons against beta amyloid toxicity." Annals of Neurology. (in press).
[Publications] Kitamura Y: "Changes of p53 in the brain of patients with Alzheimer's disease." Biochem Biophys Res Commun. (in press).
[Publications] Shimohama S: "Nicotinic α7 receptors protect against glutamate neurotoxicity and neuronal ischemic damage." Brain Research. (in press).
[Publications] Kawamata J: "Amyotrophic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Elsevier(Eds.I.Nakano & A.Hirano), 399 (1996)
[Publications] Imura T: "Amyotrophic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Elsevier(Eds.I.Nakano & A.Hirano), 399 (1996)

1996 Fiscal Year Annual Research Report

ヒト脳由来の新規神経保護活性物質の探索

Principal Investigator

下濱 俊 京都大学, 医学研究科, 助手 (60235687)

Research Products

[Publications] Sawada H: "Methyl phenylpyoldium ion(MPP^+)enbances glutamate-induced cytotoxicity against dopaminergic neurons in cultuned rat mesen cephalon" Journal of Neuroscience Research. 43. 55-62 (1996)

[Publications] Sawada H: "Different mechanisms of glutamate-induced neuronal death between dopaminergic and non-dopaminergic neurons in rat mesencephalic culture" Journal of Neuroscience Research. 43. 503-510 (1996)

[Publications] Shimohama S: "Nicotine-induced protection against glutamate cytotoxicity:Nicotinic cholinergic receptor・mediated inhibition of nitric oxide formation" Annals of New York Academy of Sciences. 777. 356-361 (1996)

[Publications] Shimohama S: "Changers of neurocaloin,a calcium-binding protein,in the brain of patients with Alzheimer's disease" Brain Research. 716. 233-236 (1996)

[Publications] Matsushima H: "Ca^<2+>-dependent and-independent protein kinase C changes in the brain of patients with Alzheimer's disease" Journal of Neurochemistry. 67. 317-323 (1996)

[Publications] Chachin M: "Assessment of protein kinase C mRNA levels in Alzheimer's disease brains" Japanese Journal of Pharmacology. 71. 175-177 (1996)

[Publications] Fujimoto S: "Zinc-ion-dependent acid phosphatase exhibits maguesium-ion-dependent myo-inositol-1-phosphatase activity" Biol.Pharm.Bull. 19. 882-885 (1996)

[Publications] Kawakami N: "Tyrosine phosphorylation and translocation of phospholipase C-γ_2 in polyrmorphonuclear leukocytes treated with pervanadate" Biochim Biophys Acta. 1314. 167-174 (1996)

[Publications] Sawada H: "Mechanism of resistance to NO-induced neurotoxicity in cultured rat dopaminergic neurons" Journal of Neuroscience Research. 46. 509-518 (1996)

[Publications] Kihara T: "Nicotinic receptor stimulation protects neurons against beta amyloid toxicity." Annals of Neurology. (in press).

[Publications] Kitamura Y: "Changes of p53 in the brain of patients with Alzheimer's disease." Biochem Biophys Res Commun. (in press).

[Publications] Shimohama S: "Nicotinic α7 receptors protect against glutamate neurotoxicity and neuronal ischemic damage." Brain Research. (in press).

[Publications] Kawamata J: "Amyotrophic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Elsevier(Eds.I.Nakano & A.Hirano), 399 (1996)

[Publications] Imura T: "Amyotrophic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Elsevier(Eds.I.Nakano & A.Hirano), 399 (1996)

下濱俊京都大学, 医学研究科, 助手 (60235687)