N-ethylmaleimide-sensitive fusion protein (NSF) is involved in central sensitization in the spinal cord through GluR2 subunit composition switch after inflammation
N-ethylmaleimide-sensitive fusion protein (NSF) is involved in central sensitization in the spinal cord through GluR2 subunit composition switch after inflammation
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DOI：
10.1111/j.1460-9568.2008.06293.x
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发表时间：
2008-06-01
影响因子：
3.4
通讯作者：
Ito, Seiji
中科院分区：
文献类型：
--
作者：
Katano, Tayo;Furue, Hidemasa;Ito, Seiji
中枢敏化，类似于海马中的长时程增强，是指在组织损伤或神经损伤后在脊髓背角中的体感神经元中建立的突触功效增加。在炎症过程中，包括谷氨酸受体在内的许多蛋白质被认为在突触后密度（PSD）中动态重组并参与持续性疼痛。足底注射完全弗氏佐剂（CFA）诱导的机械性痛觉过敏在4小时，但不是在24小时，由吲哚美辛，前列腺素合成的抑制剂抑制。为了阐明炎症性疼痛晚期所涉及的分子的性质，我们通过进行二维差异凝胶电泳分析了CFA注射前和注射后24小时从大鼠腰髓制备的PSD组分。通过MALDI-TOF MS和免疫印迹鉴定，N-乙基马来酰亚胺敏感融合蛋白（NSF）是PSD中下调的蛋白。注射CFA后24 h，随着NSF含量的降低，PSD中GluR 2和GluR 3含量降低，GluR 1含量升高。CFA注射后24 h的大鼠在体膜片钳记录显示，刺激发炎皮肤引起的背角神经元兴奋性突触后电流被向内整流，并被Philanthotoxin-433抑制60%，Philanthotoxin-433是一种选择性的钙渗透性α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）受体的抑制剂。这些结果表明，外周炎症通过晚期AMPA受体的亚单位组成转换引起脊髓中枢敏化。
Central sensitization, similar to long-term potentiation in the hippocampus, refers to the increased synaptic efficacy established in somatosensory neurons in the dorsal horn of the spinal cord following tissue injury or nerve damage. In the course of inflammation, many proteins including glutamate receptors are assumed to be dynamically reorganized in the postsynaptic density (PSD) and involved in persistent pain. Mechanical hyperalgesia induced by intraplantar injection of complete Freund's adjuvant (CFA) was inhibited at 4 h, but not at 24 h, by indomethacin, an inhibitor of prostanoid synthesis. To elucidate the nature of the molecule(s) involved in the late phase of inflammatory pain, we analysed the PSD fraction prepared from the lumbar spinal cord of rats before and 24 h after CFA injection by conducting two-dimensional differential gel electrophoresis. N-ethylmaleimide-sensitive fusion protein (NSF) was identified as a downregulated protein in the PSD by MALDI-TOF MS and immunoblotting. Concomitant with the decrease in NSF, GluR2 and GluR3 were decreased and GluR1 was conversely increased in the PSD fraction 24 h after CFA injection. In vivo patch-clamp recordings of rats 24 h after CFA injection showed that excitatory postsynaptic currents of dorsal horn neurons evoked by pinch stimuli to inflamed skin were inwardly rectified and inhibited by 60% by philanthotoxin-433, a selective inhibitor of the Ca2+-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor. These results suggest that peripheral inflammation gives rise to central sensitization in the spinal cord through subunit composition switch of AMPA receptors in the late phase.