口腔医学研究

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (9): 792-797.DOI: 10.13701/j.cnki.kqyxyj.2023.09.006

• 口腔颌面外科学研究 • 上一篇    下一篇

过表达P2X4受体介导三叉神经痛的实验研究

黄若瑜1, 宋玉丰1, 周敏1, 徐昊旻1, 刘俊1, 张明明2, 高云2, 熊伟1*   

  1. 1.江西省口腔生物医学重点实验室 江西省口腔疾病临床医学研究中心 南昌大学附属口腔医院 江西 南昌 330006;
    2.南昌大学基础医学院生理教研室 江西 南昌 330006
  • 收稿日期:2023-06-01 出版日期:2023-09-28 发布日期:2023-09-25
  • 通讯作者: *熊伟,E-mail:xiongwei96@163.com
  • 作者简介:黄若瑜(1995~ ),女,江西赣州人,硕士,住院医师,研究方向:口腔医学。
  • 基金资助:
    国家自然科学基金(编号:81860199)

Experimental Study on Overexpression of P2X4 Receptors Mediating Trigeminal Neuralgia

HUANG Ruoyu1, SONG Yufeng1, ZHOU Min1, XU Haomin1, LIU Jun1, ZHANG Mingming2, GAO Yun2, XIONG Wei1*   

  1. 1. Jiangxi Provincial Key Laboratory of Stomatology and Biomedicine, Jiangxi Clinical Research Center for Stomatology, Affiliated Stomatology Hospital of Nanchang University, Nanchang 330006, China;
    2. Department of Physiology, School of Basic Medical Sciences, Nanchang University, Nanchang 330006, China
  • Received:2023-06-01 Online:2023-09-28 Published:2023-09-25

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摘要： 目的: 本课题组前期发现,P2X4受体可能通过调控脑源性神经生长因子(brain-derived neurotrophic factor, BDNF)表达,进而介导三叉神经痛(trigeminal neural,TN)分子机制。为此,在TN大鼠模型中转染过表达P2X4受体,进行验证。方法: 以结扎框下神经慢性损伤法建立TN大鼠模型,P2X4受体过表达质粒舌下静脉转染至TN大鼠中,以面部痛敏阈检测仪、荧光定量PCR、间接免疫荧光、Western blot和ELISA法检测大鼠三叉神经脊束核 (spinal trigeminal nucleus,STN)和三叉神经节(trigeminal ganglia,TG)部位疼痛阈值,P2X4、BDNF和TrkB转录水平,P2X4和BDNF定位情况,P2X4、BDNF、TrkB蛋白表达和ERK磷酸化水平,炎性因子白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达。结果: 过表达P2X4受体组大鼠疼痛阈值降低,STN和TG部位P2X4、BDNF受体和TrkB转录水平升高(P＜0.01),STN中BDNF受体与小胶质标志物OX42共定位、TG中P2X4与胶质纤维酸性蛋白 (glial fibrillary acidic protein,GFAP)共定位,STN和TG部位ERK磷酸化水平升高(P＜0.01),大鼠血清中IL-1β和TNF-α表达水平升高(P＜0.05)。结论: 过表达P2X4受体降低大鼠疼痛阈值,表明P2X4离子参与了TN疼痛通路,其机制为P2X4调控了BDNF/ERK通路,增强了ERK磷酸化和炎性因子IL-1β和TNF-α表达。

关键词: P2X4受体, 三叉神经痛, 脑源性神经营养因子, ERK磷酸化, 炎性因子

Abstract: Objective: To validate whether P2X4 receptors mediate the molecular mechanism of trigeminal neuralgia (TN) by regulating the expression of brain-derived neurotrophic factor (BDNF) through transfecting overexpressed P2X4 receptors in a TN rat model. Methods: The P2X4 receptor overexpression plasmid was transfected into the sublingual vein of TN rats. The facial pain threshold assay, quantitative fluorescence PCR, indirect immunofluorescence, protein blotting, and ELISA were used to detect the pain thresholds in rat spinal trigeminal nucleus (STN) and trigeminal ganglia (TG), the transcript levels of P2X4, BDNF, and TrkB, the localization of P2X4 and BDNF, the protein expression of P2X4, BDNF, and TrkB, the phosphorylation levels of ERK, and the expressions of inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Results: In the P2X4 receptor overexpression group, pain thresholds were reduced, transcript levels of P2X4, BDNF receptor, and TrkB were increased in STN and TG sites (P<0.01), BDNF receptor was co-localized with microglial marker OX42 in STN, P2X4 was co-localized with glial fibrillary acidic protein (GFAP) in TG, ERK phosphorylation was increased in STN and TG sites (P<0.01), and IL-1β and TNF-α expression levels were increased in serum (P<0.01). Conclusion: Overexpression of P2X4 receptors decreased pain thresholds in rats, suggesting that P2X4 ions are involved in the TN pain pathway by the mechanism that P2X4 regulates the BDNF/ERK pathway, and enhances ERK phosphorylation and expression of inflammatory factors IL-1β and TNF-α.

Key words: P2X4 receptors, trigeminal ganglia, brain-derived neurotrophic factor, ERK phosphorylation, inflammatory factors