Regulation of neuronal cell functions by S-nitroso-cysteine

S-亚硝基半胱氨酸调节神经细胞功能

• 批准号: 11680749
• 负责人: MURAYAMA Toshihiko
• 金额: $ 2.11万
• 依托单位: Chiba University (2000)Hokkaido University (1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680749/
• 关键词: nitric oxide; NO radical; S-nitroso-cysteine; neuronal cells; phospholipase A2; arachidonic acid; apoptosis; caspase

Previously, we proposed a new signal transduction mechanism [NO radical-S-nitroso-thiols formation-S-nitrosylation of proteins]. In the present study, we obtained several new findings concerning the effects of S-nitroso-cysteine on neuronal cell functions. 1) InPC12 cells, an inhibitor of thioredoxin reductase (DNCB) caused apoptosis with caspase activation. Treatment with NO compounds including S-nitroso-cysteine inhibited the caspase activation and the apoptosis. 2) S-Nitroso-cysteine, but not other NO compounds, stimulated noradrenaline release from rat brain slices. S-Nitroso-cysteine acted as a sulfhydryl agent on proteins that regulate noradrenaline release. 3) Modification of the oxidation status of the sulfhydryl groups on the caffeine-sensitive receptors by S-nitroso-cysteine regulated Ca2+ channel activity in a reversible manner in PC12 cells. 4) S-Nitroso-cysteine regulated translocation of vesicles containing synaptophysin and SNAP-25 in PC12 cells. 5) S-Nitroso-cysteine reacted with cysteine residues in GTP-binding proteins such as Gi/Go, causing inhibition of ADP-ribosylation by pertussis toxin. 6) Arachidonic acid release from PC12 cells was inhibited by cytosolic phospholipase A2 inhibitors and by S-nitroso-cysteine, but not by other NO compounds. These findings show the regulation of neuronal cell functions by S-nitroso-cysteine.

此前，我们提出了一种新的信号转导机制[NO自由基- s -亚硝基-硫醇形成- s -亚硝基化]。在本研究中，我们获得了一些关于s -亚硝基半胱氨酸对神经细胞功能影响的新发现。1)硫氧还蛋白还原酶（DNCB）抑制剂通过caspase激活导致InPC12细胞凋亡。含s -亚硝基半胱氨酸的NO化合物可抑制caspase的激活和细胞凋亡。2) s -亚硝基半胱氨酸刺激大鼠脑切片去甲肾上腺素释放，而其他NO化合物无刺激作用。s -亚硝基半胱氨酸在调节去甲肾上腺素释放的蛋白质上起巯基作用。3) s -亚硝基半胱氨酸修饰咖啡因敏感受体上巯基的氧化状态，以可逆的方式调节PC12细胞中Ca2+通道的活性。4) s -亚硝基半胱氨酸调控PC12细胞中含有synaptophysin和SNAP-25的囊泡易位。5) s -亚硝基半胱氨酸与gtp结合蛋白如Gi/Go中的半胱氨酸残基反应，引起百日毒对adp核糖基化的抑制。6)胞质磷脂酶A2抑制剂和s -亚硝基半胱氨酸可抑制花生四烯酸在PC12细胞中的释放，而其他NO类化合物则无抑制作用。这些发现表明了s -亚硝基半胱氨酸对神经元细胞功能的调节作用。

项目成果

Naganuma, T., Maekawa, M., Murayama, T.and Nomura, Y.: "Effects of S-nitroso-cysteine on proteins that regulate exocytosis in PC12 cells ; inhibitory effects on translocation of synaptophysin and ADP-ribosylation of GTP-binding proteins."Jpn.J.Pharmacol..

Naganuma, T.、Maekawa, M.、Murayama, T. 和 Nomura, Y.：“S-亚硝基半胱氨酸对调节 PC12 细胞胞吐作用的蛋白质的影响；对突触素易位和 GTP-ADP-核糖基化的抑制作用

Maekawa,M.,: "Involvement of Hg2+-sensitive sulfhydryl groups in regulating noradrenaline release by S-nitroso-cysteine in rat brain slices."Biochem.Phannacol.. 59. 839-845 (2000)

Maekawa，M.，：“Hg2 敏感的巯基参与调节大鼠脑切片中 S-亚硝基半胱氨酸的去甲肾上腺素释放。”Biochem.Phannacol.. 59. 839-845 (2000)

Yoshinaga, N.: "Possible involvement of cytosolic phospholipase A2 in cell death induced by 1-methl-4-phenylpyridinium ion (MPP+), a dopaminergic neurotoxin, in GH3 cells"Brain Res.. (in press). (2000)

Yoshinaga, N.：“胞质磷脂酶 A2 可能参与 GH3 细胞中 1-甲基-4-苯基吡啶鎓离子 (MPP)（一种多巴胺能神经毒素）诱导的细胞死亡”Brain Res..（出版中）。

Maekawa, M.: "Involvement of Hg2+-sensitive sulfhydryl groups in regulating noradrenaline release by S-nitroso-cysteine in rat brain slices"Biochem. Pharmacol.. (in press). (2000)

Maekawa, M.：“Hg2 敏感的巯基参与调节大鼠脑切片中 S-亚硝基-半胱氨酸的去甲肾上腺素释放”Biochem。

Ishikawa, A.: "Cell death by 1-chloro-2,4-dinitrobenzene, an inhibitor of thioredoxin reductase and its dual regulation by NO"Neurosci. Lett.. (in press). (2000)

Ishikawa, A.：“1-氯-2,4-二硝基苯（一种硫氧还蛋白还原酶抑制剂）导致的细胞死亡及其受 NO 的双重调节”Neurosci。