PHOSPHODIEASERASE INHIBITION DECREASES NEURONAL

磷酸酶抑制可减少神经元

• 批准号: 12671422
• 负责人: OGATA Tadanori
• 金额: $ 1.28万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671422/
• 关键词: spinal cord; nitric oxide; neuron; cyclic GMP; phosphodiesterase; phosphodiesterase; cyclic AMP; TNF-α; アストロサイト; マイクログリア

The effect of intracellular cyclic GMP increase on neuronal damage was tested using a newly developed nitric oxide-related injury model of cultured spinal cord neurons. Neuronal damage after 24 hours' exposure to sodium nitroprusside (SNP, 6.25-100 μM), an NO donor, was evaluated by measuring the activity of released lactate dehydrogenase (LDH) from degenerated neurons. The intracellular cyclic GMP increase caused by the addition of 100 μM 8-bromo-cyclic GMP, a cell membrane permeable cyclic GMP, strongly inhibited the neuronal damage induced by 10 μM SNP, although 8-bromo-cyclic AMP yielded no effect. The addition of 8-bromo-cyclic GMP induced nerve growth factor (NGF) production in cultured neurons, suggesting that the neuroprotection via intracellular cyclic GMP increase is mediated by NGF production. The addition of propentofylline (PPF; 1-100 μM), a xanthine derivative and phosphodiesterase (PDE) inhibitor, enhanced intracellular cyclic GMP elevation induced by SNP exposure. The neuronal damage induced by 10 μM SNP exposure for 24 hours was almost completely blocked in the presence of PPF.

使用新开发的培养脊髓神经元的一氧化氮相关损伤模型测试细胞内环GMP增加对神经元损伤的影响。用一氧化氮（NO）供体硝普钠（SNP，6.25-100 μM）处理神经元24小时后，通过测定变性神经元释放的乳酸脱氢酶（LDH）活性来评价神经元的损伤。加入100 μM的8-溴环GMP（一种细胞膜可渗透的环GMP）所引起的细胞内环GMP的增加强烈地抑制了10 μM的SNP所引起的神经元损伤，而8-溴环AMP则没有这种作用。加入8-溴环GMP诱导培养神经元产生神经生长因子（NGF），表明通过细胞内环GMP增加的神经保护作用是由NGF的产生介导的。添加丙戊茶碱（PPF; 1-100 μM）（一种黄嘌呤衍生物和磷酸二酯酶（PDE）抑制剂）可增强SNP暴露诱导的细胞内环GMP升高。PPF可完全阻断10 μM SNP作用24 h引起的神经元损伤。