Objective The rostral anterior cingulate cortex(rACC)is implicated in processing the emotional component of pain.N-methyl-D-aspartate receptors(NMDARs)are highly expressed in the rACC and mediate painrelated affect by activating a signaling pathway that involves cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)and/or extracellular regulated kinase(ERK)/cAMP-response element-binding protein(CREB).The present study investigated the contributions of the NMDAR glycine site and GluN2B subunit to the activation of ERK and CREB both in vitro and in vivo in rat rACC.Methods Immunohistochemistry and Western blot analysis were used to separately assess the expression of phospho-ERK(pERK)and phospho-CREB(pCREB)in vitro and in vivo.Double immunostaining was also used to determine the colocalization of pERK and pCREB.Results Both bath application of NMDA in brain slices in vitro and intraplantar injection of formalin into the rat hindpaw in vivo induced significant up-regulation of pERK and pCREB in the rACC,which was inhibited by the NMDAR antagonist DL-2-amino-5-phospho-novaleric acid.Selective blockade of the NMDAR GluN2B subunit and the glycinebinding site,or degradation of endogenous D-serine,a co-agonist for the glycine site,significantly decreased the up-regulation of pERK and pCREB expression in the rACC.Further,the activated ERK predominantly colocalized with CREB.Conclusion Either the glycine site or the GluN2B subunit of NMDARs participates in the phosphorylation of ERK and CREB induced by bath application of NMDA in brain slices or hindpaw injection of 5% formalin in rats,and these might be fundamental molecular mechanisms underlying pain affect.
Hong CaoWen-Hua RenMu-Ye ZhuZhi-Qi ZhaoYu-Qiu Zhang
本文采用大鼠坐骨神经慢性压迫损伤引起的神经病理痛模型,研究脊髓背角细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)在外周神经损伤引起的神经病理疼痛发生中的作用。结果显示,单侧坐骨神经压迫性损伤后1天,大鼠损伤侧脊髓背角ERK的磷酸化(激活)水平显著上调,其下游转录因子cAMP反应原件结合蛋白(cAMP response element binding protein,CREB)在双侧脊髓背角的激活水平也同时上调,而此时由神经损伤引起的痛觉敏化行为尚未出现。神经损伤之前和损伤后早期鞘内给予促分裂原活化蛋白激酶激酶(mitogen-activated protein kinase kinase,MEK)的抑制剂U0126,可阻断和延迟坐骨神经损伤引起的触诱发痛和热痛觉过敏行为的发生。这些结果提示,脊髓背角ERK-CREB信号的激活参与外周神经损伤引起的神经病理疼痛的发生,对该信号通路的早期干预可能是控制神经病理性疼痛的重要手段。
