A fundamental study for application of nitric oxide to a diagnosis and treatments of the chronic pain

一氧化氮在慢性疼痛诊断和治疗中应用的基础研究

• 批准号: 18613023
• 负责人: ABE Tetsuya
• 金额: $ 2.5万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18613023/
• 关键词: nitric oxide; neuropathic pain; primary afferent; NMDA receptor; spinal cord; enzymohistochemistry; Imaging; mice; 神経因性痺痛; ノシセプチン

We established an ex vivo system to elucidate biochemical and molecular mechanisms for nNOS activation by the use of a combination of isolated intact spinal cord preparations and NADPH-diaphorase histochemistry.Nociceptin/orphanin FQ (N/OFQ) was earlier shown to be involved in the maintenance of neuropathic pain by activating neuronal nitric oxide synthase (nNOS). We examined the N/OFQ signal pathways coupled to nNOS activation in the spinal cord by using this ex vivo system. N/OFQ enhanced nNOS activity in the superficial layer of the spinal cord, as assessed by NADPH-diaphorase histochemistry, in a time- and dose-dependent manner. The maximum effect was observed at 3-10 nM. The N/OFQ-stimulated nNOS activity was inhibited by NMDA receptor antagonists MK-801 and D-AP5, but not by the NR2B-selective antagonist; and the stimulated activity was observed in NR2D(-/-) mice, but not in NR2A(-/-) or NR2A(-/-)/NR2D(-/-) mice. N/OFQ receptor antagonists attenuated the nNOS activity stimulated … More by N/OFQ, but not that by NMDA. N/OFQ stimulated nNOS activity by a biochemical cascade initiated by activation of NMDA receptors containing NR2A.To clarify whether NO itself affected nNOS activity in the spinal cord as a retrograde messenger, we examined the involvement of the NO/cGMP signaling pathway in the regulation of nNOS activity. NO-generating agents enhanced NADPH-diaphorase staining in the spinal cord 8-Br-cGMP also enhanced it similar to that by NO-generating agents, and 8-Br-cAMP and forskolin, an activator of adenylate cyclase, enhanced it moderately. NO-generating agents markedly increased the cGMP level in the spinal cord. Additionally, the nNOS activation was completely inhibited by NMDA antagonists MK-801 and d-AP5, partially by the NR2B-selective antagonist, and was attenuated in NR2A(-/-) and NR2A(-/-)/2D(-/-) mice. These results suggest that NO may regulate nNOS activity as a retrograde messenger in the spinal cord via activation of NMDA receptor containing NR2A and NR2B subunits. Less

我们建立了一个体外系统，通过结合分离的完整脊髓标本和NADPH-黄递酶组织化学来阐明nNOS激活的生化和分子机制。Nociceptin/Oranin FQ(N/OFQ)早期被证明通过激活神经元型一氧化氮合酶(NNOS)参与神经病理性疼痛的维持。我们利用这个体外实验系统研究了脊髓中与nNOS激活相关的N/OFQ信号通路。NADPH-黄递酶组织化学显示，N/OFQ以时间和剂量依赖的方式增强脊髓浅层nNOS活性。在3-10 nm处效果最好。NMDA受体拮抗剂MK-801和D-AP5可抑制N/OFQ刺激的nNOS活性，但NR2B选择性拮抗剂不能抑制其活性，NR2D(-/-)小鼠有此激活作用，而NR2A(-/-)或NR2A(-/-)/NR2D(-/-)小鼠则无此激活作用。N/OFQ受体拮抗剂减弱…刺激的一氧化氮合酶活性更多的是N/OFQ，而不是NMDA。N/OFQ通过激活含有NR2a的NMDA受体启动的生化级联反应刺激nNOS活性。为了阐明NO本身是否作为逆行信使影响脊髓nNOS活性，我们研究了NO/cGMP信号通路在nNOS活性调节中的作用。NO生成剂增强脊髓NADPH-黄递酶的染色，8-BR-cGMP与NO生成剂相似，8-BR-cAMP和腺苷环化酶激活剂Forsklin适度增强脊髓NADPH-黄递酶的表达。NO生成剂可显著增加脊髓组织中cGMP水平。NMDA拮抗剂MK-801和d-AP5可部分抑制NR2B选择性拮抗剂对nNOS的激活，而NR2A(-/-)和NR2A(-/-)/2D(-/-)小鼠的nNOS活性则明显减弱。这些结果提示，NO可能通过激活含有NR2A和NR2B亚单位的NMDA受体，作为逆行信使调节脊髓内nNOS的活性。较少

项目成果

Differentiation and migration of progenitor cells for maitenance of neuropathic pain in the model using nestin promoter-GFP transgenic mice

使用巢蛋白启动子-GFP转基因小鼠模型中维持神经性疼痛的祖细胞的分化和迁移

• 发表时间: 2007
• 作者: Matsumura;S.
• 通讯作者: S.

痛みと闘う 各科の取り組み システム論的な見方による難治性疼痛の予防と治療

对抗疼痛：各部门从系统角度防治顽固性疼痛的努力

• 发表时间: 2007
• 期刊: 医学のあゆみ 223
• 作者: 中井 吉英;水野 康行;阿部 哲也
• 通讯作者: 阿部 哲也

痛みの臨床心理学

疼痛的临床心理学

• 发表时间: 2006
• 期刊: 理学療法 23
• 作者: 阿部 哲也;中井 吉英
• 通讯作者: 中井 吉英

Nitric oxide (NO) serves as a retrograde messenger to activate neuronal NO synthase in the spinal cord via NMDA receptors

• DOI: 10.1016/j.niox.2007.04.004
• 发表时间: 2007-08-01
• 期刊: NITRIC OXIDE-BIOLOGY AND CHEMISTRY
• 影响因子: 3.9
• 作者: Xu, Li;Mabuchi, Tamaki;Ito, Seiji
• 通讯作者: Ito, Seiji

The opioid peptide nociceptin/orphanin FQ mediates prostaglandin E2‐induced allodynia, tactile pain associated with nerve injury

• DOI: 10.1111/j.1460-9568.2006.04623.x
• 发表时间: 2006-02
• 期刊: European Journal of Neuroscience
• 影响因子: 3.4
• 作者: E. Okuda‐Ashitaka;T. Minami;S. Matsumura;H. Takeshima;R. Reinscheid;O. Civelli;S. Ito