中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (1): 140-148.doi: 10.4103/1673-5374.243719

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

TRPA1激活或抑制是否影响背根神经节神经元TRPA1和CGRP的表达

  

  • 出版日期:2019-01-15 发布日期:2019-01-15
  • 基金资助:

    国家自然科学基金(81501935；基金负责人：李浩)和山东省自然科学基金(No.ZR2014HQ065；基金负责人：李浩)

Effects of TRPA1 activation and inhibition on TRPA1and CGRP expression in dorsal root ganglion neurons

Xiao-Lei Wang1, Li-Wei Cui2, Zhen Liu1, Yue-Ming Gao1, Sheng Wang1, Hao Li3, Hu-Xiang Liu1, Ling-Jia Yu1   

  1. 1 Department of Rheumatology, Qilu Hospital of Shandong University, Jinan, Shandong Province, China
    2 Department of Respiratory Medicine, Qilu Hospital of Shandong University, Jinan, Shandong Province, China
    3 Department of Orthopedics, Qilu Hospital of Shandong University, Jinan, Shandong Province, China
  • Online:2019-01-15 Published:2019-01-15
  • Contact: Ling-Jia Yu, BS, aolin88@126.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81501935 (to HL); the Natural Science Foundation of Shandong Province of China, No. ZR2014HQ065 (to HL).

PDF

296

摘要：

定位于感受伤害性刺激的初级传入背根神经节神经元的瞬时受体电位锚蛋白1(transient receptor potential ankyrin 1，TRPA1)，在疼痛和神经源性炎症发生发展和传导伤害性感觉刺激信号的过程中发挥关键的作用。神经源性炎症的产生可能与TRPA1诱发的降钙素基因相关肽(calcitonin gene-related peptide，CGRP)释放有关，然而，TRPA1和CGRP的表达是否与TRPA1的激活状态有关联目前并不清楚。实验利用体外培养的背根神经节神经元，观察了经TRPA1激动剂甲醛或阻断剂薄荷醇刺激后TRPA1和CGRP的表达变化，探讨与细胞外信号调节激酶1/2(extracellular signal-regulated protein kinase 1/2，ERK1/2)相关的作用机制。(1)将培养的背根神经节神经元分别经甲醛、薄荷醇、ERK1/2抑制剂PD98059+甲醛以及PD98059+薄荷醇刺激后，用real-time PCR、Western blot和免疫荧光双标技术检测了TRPA1和CGRP在mRNA和蛋白水平的表达以及在背根神经节神经元原位的表达；(2)结果显示，甲醛刺激可上调背根神经节神经元TRPA1和CGRP mRNA和蛋白的表达，并使TRPA1和CGRP免疫反应阳性神经元比例增加，而薄荷醇的调节作用则呈现出与以上相反的趋势；(3)上述数据说明，TRPA1激动剂甲醛对背根神经节神经元CGRP表达的影响作用可被ERK1/2抑制剂PD98059所阻断，而PD98059不影响甲醛和TRPA1阻断剂薄荷醇对TRPA1表达的调节作用。

orcid: 0000-0002-4786-2666(Ling-Jia Yu)

关键词: TRPA 1, TRPV 1, 甲醛, 薄荷脑, CGRP, 背根神经节, 神经元, 神经源性炎症, 伤害性刺激信号, ERK 1/2, 神经再生

Abstract:

Transient receptor potential ankyrin 1 (TRPA1) is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion (DRG) neurons. TRPA1 plays a major role in the transmission of nociceptive sensory signals. The generation of neurogenic inflammation appears to involve TRPA1-evoked release of calcitonin gene-related peptide (CGRP). However, it remains unknown whether TRPA1 or CGRP expression is affected by TRPA1 activation. Thus, in this study, we examined TRPA1 and CGRP expression in DRG neurons in vitro after treatment with the TRPA1 activator formaldehyde or the TRPA1 blocker menthol. In addition, we examined the role of extracellular signal-regulated protein kinase 1/2 (ERK1/2) in this process. DRG neurons in culture were exposed to formaldehyde, menthol, the ERK1/2 inhibitor PD98059 + formaldehyde, or PD98059 + menthol. After treatment, real-time polymerase chain reaction, western blot assay and double immunofluorescence labeling were performed to evaluate TRPA1 and CGRP expression in DRG neurons. Formaldehyde elevated mRNA and protein levels of TRPA1 and CGRP, as well as the proportion of TRPA1- and CGRP-positive neurons. In contrast, menthol reduced TRPA1 and CGRP expression. Furthermore, the effects of formaldehyde, but not menthol, on CGRP expression were blocked by pretreatment with PD98059. PD98059 pretreatment did not affect TRPA1 expression in the presence of formaldehyde or menthol.

Key words: nerve regeneration, TRPA1, TRPV1, formaldehyde, menthol, CGRP, dorsal root ganglion, neuron, neurogenic inflammation, nociceptive signal, ERK1/2, neural regeneration