Increasing evidence suggests that transmembrane protein 16A (TMEM16A) in nociceptive neurons is an important molecular component contributing to peripheral pain transduction. The present study aimed to evaluate the role and mechanism of TMEM16A in chronic nociceptive responses elicited by spared nerve injury (SNI). In this study, SNI was used to induce neuropathic pain. Drugs were administered intrathecally. The expression and cellular localization of TMEM16A, the ERK pathway, and NK-1 in the dorsal root ganglion (DRG) were detected by western blot and immunofluorescence. Behavioral tests were used to evaluate the role of TMEM16A and p-ERK in SNI-induced persistent pain and hypersensitivity. The role of TMEM16A in the hyperexcitability of primary nociceptor neurons was assessed by electrophysiological recording. The results show that TMEM16A, p-ERK, and NK-1 are predominantly expressed in small neurons associated with nociceptive sensation. TMEM16A is colocalized with p-ERK/NK-1 in DRG. TMEM16A, the MEK/ERK pathway, and NK-1 are activated in DRG after SNI. ERK inhibitor or TMEM16A antagonist prevents SNI-induced allodynia. ERK and NK-1 are downstream of TMEM16A activation. Electrophysiological recording showed that CaCC current increases and intrathecal application of T16Ainh-A01, a selective TMEM16A inhibitor, reverses the hyperexcitability of DRG neurons harvested from rats after SNI. We conclude that TMEM16A activation in DRG leads to a positive interaction of the ERK pathway with activation of NK-1 production and is involved in the development of neuropathic pain after SNI. Also, the blockade of TMEM16A or inhibition of the downstream ERK pathway or NK-1 upregulation may prevent the development of neuropathic pain.

越来越多的证据表明，伤害性神经元中的跨膜蛋白 16A (TMEM16A) 是促进外周疼痛转导的重要分子成分。本研究旨在评估 TMEM16A 在幸免神经损伤 (SNI) 引起的慢性伤害性反应中的作用和机制。在这项研究中，SNI 用于诱导神经性疼痛。药物被鞘内给药。通过蛋白质印迹和免疫荧光检测TMEM16A、ERK通路和NK-1在背根神经节（DRG）中的表达和细胞定位。行为测试用于评估 TMEM16A 和 p-ERK 在 SNI 引起的持续性疼痛和超敏反应中的作用。通过电生理记录评估 TMEM16A 在初级伤害感受器神经元过度兴奋中的作用。结果表明，TMEM16A，p-ERK 和 NK-1 主要在与伤害性感觉相关的小神经元中表达。TMEM16A 与 DRG 中的 p-ERK/NK-1 共定位。TMEM16A、MEK/ERK 通路和 NK-1 在 SNI 后在 DRG 中被激活。ERK 抑制剂或 TMEM16A 拮抗剂可防止 SNI 诱导的异常性疼痛。ERK 和 NK-1 是 TMEM16A 激活的下游。电生理记录显示，CaCC 电流增加和 T16Ainh-A01（一种选择性 TMEM16A 抑制剂）的鞘内应用可逆转 SNI 后从大鼠采集的 DRG 神经元的过度兴奋。我们得出结论，DRG 中的 TMEM16A 激活导致 ERK 通路与 NK-1 产生激活的正相互作用，并参与 SNI 后神经性疼痛的发展。还，