Role of Nitric Oxide in neuronal death and its modulation by neuroprotective factors

一氧化氮在神经元死亡中的作用及其神经保护因子的调节

• 批准号: 07457539
• 负责人: AKAIKE Akinori
• 金额: $ 4.74万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457539/
• 关键词: Alzheimer's Disease; Nitric Oxide; Glutamate; Neuronal Death; Cortex; Nicotine; Culture; Phospholipase C-delta; アセチルコリン受容体; 初代培養神経細胞; NMDA受容体; アセチルコリン; α-ブンガロトキシン; 細胞死; ニューロン

The purpose of this study was to elucidate the mechanism underlying neuronal death in neurodegeneration diseases in central nervous system (CNS), such as cerebrovascular disease and Alzheimer's disease. We studied the mechanisms underlying glutamate neurotoxicity mediated by nitric oxide (NO) and protective effect of nicotine against glutamate neurotoxicity using cultured rat cortical neurons. 1.We examined the protective effect on nicotine. The cell viability was markedly reduced when cultures were briefly exposed to glutamate or N-methyl-D-aspartate (NMDA). Incubating the cultures with nicotine (10muM) prior to glutamate exposure reduced its cytotoxicity. The neuroprotective effect of nicotine against glutamate neurotoxicity was antagonized by hexamethonium, an antagonist specific to nicotinic acetylcholine receptor. Both brief exposures to ionomycin, a calcium ionophore, and S-nitrosocysteine (SNOC), an NO donor, induced delayd neuronal death. Nicotine prevented ionomycin-induced cy … More totoxicity without affecting SNOC-induced cytotoxicity. These results suggest that the nicotinic receptor stimulation induces neuroprotection against glutamate cytotoxicity by reducing NO-formation. 2.We have previously shown that anti-phospholipase C-delta (PLC-delta) antibody produced intense staining of neurofibrillary tangles, the neurite surrounding senile plaque cores and neuropil threads in the brain of patients with Alzheimer's disease. We examined the influence of glutamate on PLC-delta immunoreactivity in the cultures using anti-PLC-delta antibody. Exposure to glutamate exhibited increased immunostaining with the anti-PLC-delta antibody. The increase in PLC-delta immunoreactivity was prevented by both application of MK-801, an NMDA receptor antagonist, and N^<omega>-nitro-L-arginine, an NO synthase inhibitor. These results suggest that NO formation secondary to NMDA receptor activation by glutamate leads to similar modifications of PLC-delta to those seen in Alzheimer's disease. The results in this study will offer basic materials to drive forward developmental research for neuroprotective drugs. Less

本研究的目的是阐明中枢神经系统（CNS）神经退行性疾病（如脑血管疾病和阿尔茨海默病）中神经元死亡的机制。我们使用培养的大鼠皮层神经元研究了一氧化氮 (NO) 介导的谷氨酸神经毒性的机制以及尼古丁对谷氨酸神经毒性的保护作用。 1.我们检查了对尼古丁的保护作用。当培养物短暂暴露于谷氨酸或 N-甲基-D-天冬氨酸 (NMDA) 时，细胞活力显着降低。在接触谷氨酸之前用尼古丁 (10μM) 孵育培养物可降低其细胞毒性。烟碱对谷氨酸神经毒性的神经保护作用被六甲铵（一种烟碱乙酰胆碱受体特异性拮抗剂）所拮抗。短暂暴露于离子霉素（一种钙离子载体）和 S-亚硝基半胱氨酸（SNOC）（一种 NO 供体）都会导致神经元延迟死亡。尼古丁可防止离子霉素诱导的细胞毒性，而不影响 SNOC 诱导的细胞毒性。这些结果表明烟碱受体刺激通过减少NO形成而诱导针对谷氨酸细胞毒性的神经保护。 2.我们之前已经证明，抗磷脂酶 C-delta (PLC-delta) 抗体会对阿尔茨海默病患者大脑中的神经原纤维缠结、老年斑核心周围的神经突和神经纤维网线产生强烈的染色。我们使用抗 PLC-delta 抗体检查了谷氨酸对培养物中 PLC-delta 免疫反应性的影响。暴露于谷氨酸会增加抗 PLC-delta 抗体的免疫染色。 PLC-δ免疫反应性的增加通过使用MK-801(一种NMDA受体拮抗剂)和N^O-硝基-L-精氨酸(一种NO合酶抑制剂)来阻止。这些结果表明，谷氨酸激活 NMDA 受体继发 NO 形成，导致 PLC-δ 发生与阿尔茨海默病中所见类似的修饰。本研究的结果将为推动神经保护药物的开发研究提供基础材料。较少的

项目成果

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