1999 Fiscal Year Annual Research Report

新規カルモデュリン拮抗薬の抗アポトーシス作用の神経薬理学的研究

Research Project

Project/Area Number	11877018
Research Institution	Kumamoto University
Principal Investigator	福永浩司熊本大学, 医学部, 助教授 (90136721)
Co-Investigator(Kenkyū-buntansha)	宮本英七熊本大学, 医学部, 教授 (50109659)
Keywords	カルモデュリン / アポトーシス / 脳虚血 / CaMキナーゼII / 一酸化窒素合成酵素 / アストロサイト / 一酸化窒素 / 神経保護薬
Research Abstract	新規カルモデュリン拮抗薬であるDY-9760eはin vitroにおいて種々のカルモデュリン依存性酵素を阻害する。白崎(第一製薬)はラット中大脳動脈閉塞モデルにおいて虚血に伴う神経細胞障害を有意に抑制することを報告した。本研究では神経細胞死におけるカルモデュリンの役割を明らかにすることによって、カルモデュリン拮抗薬の神経細胞保護薬としての可能性を追究する。DY-9760eはin vitroにおいてカルモデュリン依存性酵素、特に、CaMキナーゼIIとカルモデュリン依存性一酸化窒素合成酵素(NOS)を強く阻害した。NOSにおいては内皮由来および神経由来NOSの両者を同程度に阻害するのに対してマクロファージに存在する誘導型NOSに対しては阻害効果を示さなかった。次に、細胞系におけるDY-9760eの効果を検討した。神経芽腫細胞でカルシウム刺激によるNO産生を1μMで有意に阻害した。これに対して、神経細胞におけるCaMキナーゼIIの活性化反応は1μMではほとんど影響されなかった。同様に、カルモデュリン依存性プロテインホスファターゼ、アデニル酸シクラーゼに対しても阻害効果を示さなかった。以上の結果からDY-9760eは細胞系ではカルモデュリン依存性NOSに対して選択的な阻害効果を持つことが明らかとなった。脳虚血モデルにおける神経細胞保護効果はNOSの阻害効果に基づくものであると考えられる。DY-9760eはアストロサイトにおいてカルシウムパラドックスによるアポトーシスを抑制した。アストロサイトにおいてはNOSの発現が低いことからDY-9760eの新たな標的分子が存在する可能性がある。今後は、アストロサイトにおけるDY-9760eの抗アポトーシス作用のメカニズムについて追究する。

Research Products

(17 results)

All Other

All Publications (17 results)

[Publications] M. Ohmitsu: "Phosphorylation of myristoylated alanine-rich protein kinase C substrate by mitogen-activated protein kinase in cultured rat hippocampal neurons following stmulation of glutamate receptors."J. Biol. Chem.. 274. 408-417 (1999)
[Publications] K. Matsumoto: "Cloning from insulinoma cells of synapsin I associated with insulin secretory granules."J. Biol. Chem.. 274. 2053-2059 (1999)
[Publications] N. Masuko: "Interaction of NE-dlg/SPA90:A possible regulatory role in molecular clustering at synaptic sites."J. Biol. Chem.. 274. 5782-5790 (1999)
[Publications] J. Kasahara: "Differential effects of a calcineurin inhibitor on gultamate-induced phosphorylation of Ca^<2+>/calmodulin-dependent protein kinases in cultured rat hippocampal neurons."J. Biol. Chem.. 274. 9061-9067 (1999)
[Publications] H. Kuwahara: "A novel NE-dlg/SPA102-associated protein, P51-nedasin, related to the amidohydorlase superfamily, interferes with the association between NE-dlg/SAP102 and N-Methy1-D-aspartate receptor."J. Biol. Chem.. 274. 32204-32214 (1999)
[Publications] J. Liu: "Differential roles of Ca^<2+>/calmodulin-dependent protein kinases II and mitogen-activated protein dinase activation in hippocampal long-term potentiation."J. Neurosci.. 19. 8292-8299 (1999)
[Publications] M. Morioka: "Activities of calcineurin and phosphatase 2A in the hippocampus after transient forebrain ischemia."Brain Res.. 828. 135-144 (1999)
[Publications] H. Kanasaki: "Mitogen-activated protein kinase activation by stimulation with thyrotropin-releasing hormone in rat pituitary GH3 cells."Biol. of Reprod.. 61. 319-325 (1999)
[Publications] I. Munir: "Mitogen-activated protein kinase activation and regualtion of cycloxygenase 2 expression by platelet-actevating factor and hCG in human endometrial adenocarcinoma cell line HEC-1B"J. Reprod. Fertil.. 117. 49-59 (1999)
[Publications] Q. Zhang: "Changes of free fatty acids and acyl-coas in rat brain hippocampal slice with tetraehylammonium-induced long-tem potentiation."Biochem. Biophys. Res. Commun.. 267. 208-212 (1999)
[Publications] K. Fukunaga: "Current studies on a working model of CaM kinase II in hippocampal long-term potentiation and memory."Jpn. J. Pharmacol.. 79. 7-15 (1999)
[Publications] 福永浩司: "海馬における神経活動依存性シナプス伝達効率の調節"蛋白質核酸酵素、増刊「神経回路形成と機能発達」. 45. 474-482 (2000)
[Publications] J. Kasahara: "Activation of CA^<2+>/calmodulin-dependent protein kinase IV in cultured rat nippocampal neurons."J. Neurosci. Res.. 59. 594-600 (2000)
[Publications] K. Jukunaga: "Decreased protein phosphatase 2A activity n hippocampal long-teme potentiation."J. Neurochem.. (in press). (2000)
[Publications] K. Fukunaga: "Ingibition of neuronal nitric oxide synthase activity by DY-9760e, anovel neuroprotective agent, in vitro and in cultured neuroblastoma cells in situ."Biochem. Pharmacol.. (in press). (2000)
[Publications] H. Kanasaki: "Involvement of p38 mitogen-activated protein kinase activation in bromocriptine-induced apoptosis in rat pitutary GH3 cells."Biol. of Reprod.. (in press). (2000)
[Publications] 宮本英七: "脳と神経:分子神経生物科学入門(編集:金子章道、川村光毅、植村慶一)第3章神経情報の統合、5.脳可塑性"共立出版. 7 (1999)

1999 Fiscal Year Annual Research Report

新規カルモデュリン拮抗薬の抗アポトーシス作用の神経薬理学的研究

Principal Investigator

福永 浩司 熊本大学, 医学部, 助教授 (90136721)

Research Products

[Publications] M. Ohmitsu: "Phosphorylation of myristoylated alanine-rich protein kinase C substrate by mitogen-activated protein kinase in cultured rat hippocampal neurons following stmulation of glutamate receptors."J. Biol. Chem.. 274. 408-417 (1999)

[Publications] K. Matsumoto: "Cloning from insulinoma cells of synapsin I associated with insulin secretory granules."J. Biol. Chem.. 274. 2053-2059 (1999)

[Publications] N. Masuko: "Interaction of NE-dlg/SPA90:A possible regulatory role in molecular clustering at synaptic sites."J. Biol. Chem.. 274. 5782-5790 (1999)

[Publications] J. Kasahara: "Differential effects of a calcineurin inhibitor on gultamate-induced phosphorylation of Ca^<2+>/calmodulin-dependent protein kinases in cultured rat hippocampal neurons."J. Biol. Chem.. 274. 9061-9067 (1999)

[Publications] H. Kuwahara: "A novel NE-dlg/SPA102-associated protein, P51-nedasin, related to the amidohydorlase superfamily, interferes with the association between NE-dlg/SAP102 and N-Methy1-D-aspartate receptor."J. Biol. Chem.. 274. 32204-32214 (1999)

[Publications] J. Liu: "Differential roles of Ca^<2+>/calmodulin-dependent protein kinases II and mitogen-activated protein dinase activation in hippocampal long-term potentiation."J. Neurosci.. 19. 8292-8299 (1999)

[Publications] M. Morioka: "Activities of calcineurin and phosphatase 2A in the hippocampus after transient forebrain ischemia."Brain Res.. 828. 135-144 (1999)

[Publications] H. Kanasaki: "Mitogen-activated protein kinase activation by stimulation with thyrotropin-releasing hormone in rat pituitary GH3 cells."Biol. of Reprod.. 61. 319-325 (1999)

[Publications] I. Munir: "Mitogen-activated protein kinase activation and regualtion of cycloxygenase 2 expression by platelet-actevating factor and hCG in human endometrial adenocarcinoma cell line HEC-1B"J. Reprod. Fertil.. 117. 49-59 (1999)

[Publications] Q. Zhang: "Changes of free fatty acids and acyl-coas in rat brain hippocampal slice with tetraehylammonium-induced long-tem potentiation."Biochem. Biophys. Res. Commun.. 267. 208-212 (1999)

[Publications] K. Fukunaga: "Current studies on a working model of CaM kinase II in hippocampal long-term potentiation and memory."Jpn. J. Pharmacol.. 79. 7-15 (1999)

[Publications] 福永浩司: "海馬における神経活動依存性シナプス伝達効率の調節"蛋白質核酸酵素、増刊「神経回路形成と機能発達」. 45. 474-482 (2000)

[Publications] J. Kasahara: "Activation of CA^<2+>/calmodulin-dependent protein kinase IV in cultured rat nippocampal neurons."J. Neurosci. Res.. 59. 594-600 (2000)

[Publications] K. Jukunaga: "Decreased protein phosphatase 2A activity n hippocampal long-teme potentiation."J. Neurochem.. (in press). (2000)

[Publications] K. Fukunaga: "Ingibition of neuronal nitric oxide synthase activity by DY-9760e, anovel neuroprotective agent, in vitro and in cultured neuroblastoma cells in situ."Biochem. Pharmacol.. (in press). (2000)

[Publications] H. Kanasaki: "Involvement of p38 mitogen-activated protein kinase activation in bromocriptine-induced apoptosis in rat pitutary GH3 cells."Biol. of Reprod.. (in press). (2000)

[Publications] 宮本英七: "脳と神経:分子神経生物科学入門(編集:金子章道、川村光毅、植村慶一)第3章 神経情報の統合、5.脳可塑性"共立出版. 7 (1999)

福永浩司熊本大学, 医学部, 助教授 (90136721)

[Publications] 宮本英七: "脳と神経:分子神経生物科学入門(編集:金子章道、川村光毅、植村慶一)第3章神経情報の統合、5.脳可塑性"共立出版. 7 (1999)