MAPK-ERK-CREB 信号通路上调奥沙利铂诱导的大鼠神经病理性疼痛中的 Nav1.6。
MAPK-ERK-CREB signaling pathway upregulates Nav1.6 in oxaliplatin-induced neuropathic pain in the rat.
机构信息
Department of Human Anatomy, School of Basic Medicine, Zhengzhou University, Zhengzhou 450001, China; Institute of Neuroscience, Zhengzhou University, Zhengzhou 450052, China.
Department of Human Anatomy, School of Basic Medicine, Zhengzhou University, Zhengzhou 450001, China; Institute of Neuroscience, Zhengzhou University, Zhengzhou 450052, China; Xinyang Central Hospital, Xinyang, China.
出版信息
Toxicol Lett. 2023 Aug 1;384:149-160. doi: 10.1016/j.toxlet.2023.07.010. Epub 2023 Jul 13.
The voltage-gated sodium channel subtype Nav1.6 is involved in the electrophysiological changes of primary sensory neurons that occur in oxaliplatin-induced neuropathic pain, but its regulatory mechanism remains unclear. In this study, Western blot, RT-qPCR, immunofluorescence staining, chromatin immunoprecipitation were used to prove the mechanism of MAPK-ERK-CREB signaling pathway participating in oxaliplatin-induced neuropathic pain by regulating Nav1.6. The results showed that p-Raf1 and p-ERK, key molecules in MAPK/ERK pathway, and Nav1.6 were significantly increased in DRGs of oxaliplatin-induced neuropathic pain rats. Inhibition of p-Raf1 and p-ERK respectively not only reduced the expression of Nav1.6 protein in DRGs of OXA rats, but also caused a decrease in Nav1.6 mRNA, which led us to further explore the transcription factor CREB regulated by MAPK/ERK pathway. Results showed that CREB was co-distributed with Nav1.6. Inhibition of CREB resulted in decreased mRNA and protein expression of Nav1.6, and alleviated oxaliplatin-induced neuropathic pain. A chromatin immunoprecipitation experiment proved that OXA caused p-CREB to directly bind to the promoter region of Scn8A, which is the encoding gene for Nav1.6, and promote the transcription of Scn8A. In summary, in this study, we found that oxaliplatin can activate the MAPK/ERK pathway, which promotes the expression and activation of CREB and leads to an increase in Scn8A transcription, and then leads to an increase in Nav1.6 protein expression to enhance neuronal excitability and cause pain. This study provides an experimental basis for the molecular mechanism of sodium channel regulation in oxaliplatin-induced neuropathic pain.
电压门控钠通道亚型 Nav1.6 参与奥沙利铂诱导的神经病理性疼痛中初级感觉神经元的电生理变化,但调节机制尚不清楚。在这项研究中,使用 Western blot、RT-qPCR、免疫荧光染色、染色质免疫沉淀来证明 MAPK-ERK-CREB 信号通路通过调节 Nav1.6 参与奥沙利铂诱导的神经病理性疼痛的机制。结果表明,MAPK/ERK 通路中的关键分子 p-Raf1 和 p-ERK 以及 Nav1.6 在奥沙利铂诱导的神经病理性疼痛大鼠的 DRG 中显著增加。p-Raf1 和 p-ERK 的抑制分别不仅降低了 OXA 大鼠 DRG 中 Nav1.6 蛋白的表达,而且导致 Nav1.6 mRNA 减少,这使我们进一步探索由 MAPK/ERK 通路调节的转录因子 CREB。结果表明 CREB 与 Nav1.6 共分布。抑制 CREB 导致 Nav1.6 的 mRNA 和蛋白表达减少,并缓解奥沙利铂诱导的神经病理性疼痛。染色质免疫沉淀实验证明,OXA 导致 p-CREB 直接结合 Nav1.6 的编码基因 Scn8A 的启动子区域,促进 Scn8A 的转录。总之,在这项研究中,我们发现奥沙利铂可以激活 MAPK/ERK 通路,促进 CREB 的表达和激活,导致 Scn8A 转录增加,进而导致 Nav1.6 蛋白表达增加,增强神经元兴奋性并引起疼痛。这项研究为奥沙利铂诱导的神经病理性疼痛中钠通道调节的分子机制提供了实验依据。