Spinal SIRPα1-SHP2 interaction regulates spinal nerve ligation-induced neuropathic pain via PSD-95-dependent NR2B activation in rats
Spinal SIRPα1-SHP2 interaction regulates spinal nerve ligation-induced neuropathic pain via PSD-95-dependent NR2B activation in rats
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DOI：
10.1016/j.pain.2012.02.006
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发表时间：
2012-05-01
期刊：
影响因子：
7.4
通讯作者：
Lin, Tzer-Bin
中科院分区：
文献类型：
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作者：
Peng, Hsien-Yu;Chen, Gin-Den;Lin, Tzer-Bin
事实上，神经性疼痛的机制没有得到很好的理解是一个主要的障碍，在发展有效的临床治疗。我们研究了信号调节蛋白α 1（SIRP α 1）和含Src同源2结构域的蛋白酪氨酸磷酸酶2（SHP 2）之间的相互作用以及下游脊髓SHP 2/突触后密度95（PSD-95）/N-甲基-D-天冬氨酸受体NR 2B亚基信号级联反应是否在神经病理性疼痛中发挥作用。脊神经结扎（L5）后，我们使用von Frey细丝测试评估触觉异常性疼痛，并通过Western印迹、逆转录聚合酶链反应、免疫共沉淀和免疫荧光分析背角样本（L4-5）。神经结扎诱导异常性疼痛、SIRP α 1、SHP 2、磷酸化SHP 2（pSHP 2）和磷酸化NR 2B（pNR 2B）表达，以及同侧背角中的SHP 2-PSD-95、pSHP 2-PSD-95、PSD-95-NR 2B和PSD-95-pNR 2B共免疫沉淀。在触诱发痛大鼠中，损伤诱导的SHP 2免疫反应位于同侧背角神经元，并且与PSD-95和NR 2B免疫反应一致。SIRP α 1沉默使用小干扰RNA（siRNA; 1，3，或5 μ g/大鼠7天）防止损伤诱导的异常性疼痛和相关的蛋白质表达，磷酸化和免疫共沉淀的变化。鞘内给予NSC-87877（一种SHP 2拮抗剂; 1、10或100 μ M/大鼠）和SIRP α 1中和抗体（1、10或30 μ g/大鼠）抑制脊神经结扎诱导的异常性疼痛、脊髓SHP 2和NR 2B磷酸化以及SHP 2/磷酸化SHP 2-PSD-95和PSD-95-NR 2B/磷酸化NR 2B共沉淀。SHP 2 siRNA导致与NSC-87877和SIRP α 1抗体治疗相似的效果，除了它阻止异常性疼痛相关的脊髓SHP 2表达。总之，我们的研究结果表明，脊髓SIRP α 1-SHP 2相互作用存在，随后触发SHP 2/PSD-95/NR 2B信号传导，从而在神经性疼痛的发展中发挥作用。(C)2012年国际疼痛研究协会。由Elsevier B出版。V.保留所有权利。
The fact that neuropathic pain mechanisms are not well understood is a major impediment in the development of effective clinical treatments. We examined whether the interaction between signal regulatory protein alpha 1 (SIRP alpha 1) and Src homology-2 domain-containing protein tyrosine phosphatase 2 (SHP2), and the downstream spinal SHP2/postsynaptic density 95 (PSD-95)/N-methyl-D-aspartate receptor NR2B subunit signaling cascade play a role in neuropathic pain. Following spinal nerve ligation (L5), we assessed tactile allodynia using the von Frey filament test and analyzed dorsal horn samples (L4-5) by Western blotting, reverse transcription polymerase chain reaction, coimmunoprecipitation, and immunofluorescence. Nerve ligation induced allodynia, SIRP alpha 1, SHP2, phosphorylated SHP2 (pSHP2), and phosphorylated NR2B (pNR2B) expression, and SHP2-PSD-95, pSHP2-PSD-95, PSD-95-NR2B, and PSD-95-pNR2B coimmunoprecipitation in the ipsilateral dorsal horn. In allodynic rats, injury-induced SHP2 immunoreactivity was localized in the ipsilateral dorsal horn neurons and coincident with PSD-95 and NR2B immunoreactivity. SIRP alpha 1 silencing using small interfering RNA (siRNA; 1, 3, or 5 mu g/rat for 7 days) prevented injury-induced allodynia and the associated changes in protein expression, phosphorylation, and coimmunoprecipitation. Intrathecal administration of NSC-87877 (an SHP2 antagonist; 1, 10, or 100 mu M/rat) and SIRP alpha 1-neutralizing antibodies (1, 10, or 30 mu g/rat) suppressed spinal nerve ligation-induced allodynia, spinal SHP2 and NR2B phosphorylation, and SHP2/phosphorylated SHP2-PSD-95 and PSD-95-NR2B/phosphorylated NR2B coprecipitation. SHP2 siRNA led to similar effects as the NSC-87877 and SIRP alpha 1 antibody treatments, except it prevented the allodynia-associated spinal SHP2 expression. In conclusion, our results suggest that a spinal SIRP alpha 1-SHP2 interaction exists that subsequently triggers SHP2/PSD-95/NR2B signaling, thereby playing a role in neuropathic pain development. (C) 2012 International Association for the Study of Pain. Published by Elsevier B. V. All rights reserved.