Disruption of nNOS-PSD95 protein-protein interaction inhibits acute thermal hyperalgesia and chronic mechanical allodynia in rodents
Disruption of nNOS-PSD95 protein-protein interaction inhibits acute thermal hyperalgesia and chronic mechanical allodynia in rodents
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DOI：
10.1111/j.1476-5381.2009.00300.x
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发表时间：
2009-09-01
影响因子：
7.3
通讯作者：
Lai, Y.
中科院分区：
文献类型：
--
作者：
Florio, S. K.;Loh, C.;Lai, Y.
背景与目的：突触后密度蛋白95 （PSD95）包含3个PSD95/犬椎间盘大/ZO-1同源结构域，连接神经元一氧化氮合酶（nNOS）与n -甲基- d -天冬氨酸（NMDA）受体。本报告评估了nNOS和PSD95蛋白-蛋白相互作用的破坏对痛觉过敏和神经性疼痛啮齿动物模型疼痛敏感性的影响。实验方法：我们生成了两个干扰nNOS-PSD95相互作用的分子：IC87201，一个小分子抑制剂；和tat-nNOS（残基1-299），一种含有nNOS的PSD95结合域的细胞渗透性融合蛋白。然后，我们使用体外和体内急性痛觉过敏和慢性异常性疼痛模型来表征这些抑制剂，这两种疾病都被认为需要nNOS激活。关键结果：IC87201和dat -nNOS（1-299）抑制了nNOS与PSD95的体外结合，但不抑制nNOS的催化活性。这两种抑制剂还阻断了nmda诱导的原代海马培养物中3',5'-环鸟苷单磷酸（cGMP）的产生。鞘内给药两种抑制剂均能有效逆转nmda诱导的小鼠热痛觉过敏。在抗痛觉过敏剂量下，对急性疼痛阈值或运动协调没有影响。鞘内注射IC87201和stat - nnos也能逆转坐骨神经慢性收缩引起的机械性异常痛。结论和意义：nNOS-PSD95相互作用在急性和慢性疼痛中维持超敏反应是重要的。破坏nNOS-PSD95相互作用提供了一种获得选择性抗痛觉化合物的新途径。
Background and purpose:Post-synaptic density protein 95 (PSD95) contains three PSD95/Dosophilia disc large/ZO-1 homology domains and links neuronal nitric oxide synthase (nNOS) with the N-methyl-D-aspartic acid (NMDA) receptor. This report assesses the effects of disruption of the protein-protein interaction between nNOS and PSD95 on pain sensitivity in rodent models of hyperalgesia and neuropathic pain.Experimental approach:We generated two molecules that interfered with the nNOS-PSD95 interaction: IC87201, a small molecule inhibitor; and tat-nNOS (residues 1-299), a cell permeable fusion protein containing the PSD95 binding domain of nNOS. We then characterized these inhibitors using in vitro and in vivo models of acute hyperalgesia and chronic allodynia, both of which are thought to require nNOS activation.Key results:IC87201 and tat-nNOS (1-299) inhibited the in vitro binding of nNOS with PSD95, without inhibiting nNOS catalytic activity. Both inhibitors also blocked NMDA-induced 3',5'-cyclic guanosine monophosphate (cGMP) production in primary hippocampal cultures. Intrathecal administration of either inhibitor potently reversed NMDA-induced thermal hyperalgesia in mice. At anti-hyperalgesic doses, there was no effect on acute pain thresholds or motor coordination. Intrathecal administration of IC87201 and tat-nNOS also reversed mechanical allodynia induced by chronic constriction of the sciatic nerve.Conclusions and implications:nNOS-PSD95 interaction is important in maintaining hypersensitivity in acute and chronic pain. Disruption of the nNOS-PSD95 interaction provides a novel approach to obtain selective anti-hyperalgesic compounds.