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Activation of P2X4 receptors in midbrain cerebrospinal fluid-contacting nucleus leads to mechanical hyperalgesia in chronic constriction injury rats

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Abstract

Neuropathic pain is a refractory pain state, and its mechanism is still not clear. Previous studies have shown that the purine receptor P2X4R expressed on hyperactive microglia in the spinal cord is essential for the occurrence and development of neuropathic pain. The cerebrospinal fluid-contacting nucleus (CSF-contacting nucleus) in the midbrain has been found to play an important role in the descending inhibition system of modulation. However, there have been no studies on P2X4R in the CSF-contacting nucleus involved in neuropathic pain. To investigate whether P2X4R is expressed in the CSF-contacting nucleus and whether its expression in the CSF-contacting nucleus is involved in the regulation of neuropathic pain, we used a model of chronic sciatic nerve ligation injury (CCI) to simulate neuropathic pain conditions. Immunohistochemistry experiments were conducted to identify the expression of P2X4R in the CSF-contacting nuclei in CCI rats, and western blot analysis showed a significant increase in P2X4R levels 7 days after modeling. Then, we packaged a P2rx4 gene-targeting shRNA in scAAV9 to knock down the P2X4R level in the CSF-contacting nucleus, and we found that CCI-induced mechanical hyperalgesia was reversed. In conclusion, P2X4R expressed in the CSF-contacting nucleus is involved in the process of neuropathic pain, and downregulating P2X4R protein in the CSF-contacting nucleus can reverse the occurrence and development of hyperalgesia, which could represent a potent therapeutic strategy for neuropathic pain.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (grant numbers 81603700, 81703898 and 81904283) and Research project of Shanghai Municipal Health Commission (No.20194Y0307).

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Author notes

  1. Wei Song, Yue Yong, and Yalan Zhou contributed equally to this work.

Authors and Affiliations

  1. Department of Anesthesiology, Shuguang Hospital Affiliated With Shanghai University of Traditional Chinese Medicine, No.528 Zhangheng Road, Shanghai, 201203, People’s Republic of China

    Wei Song, Yalan Zhou, Guijie Yu, Wei Tang, Jian Wang, Jun Guo, Lili Li & Jiangang Song

  2. Research Institute of Acupuncture Anesthesia, Shuguang Hospital Affiliated With Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China

    Yue Yong & Liyue Lu

  3. Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical University, No.209 Tongshan Road, Xuzhou, 221004, People’s Republic of China

    Licai Zhang

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Contributions

JS, LZ, and WS: conception, supervision, and design of this article. WS, YY, and YZ: experiment and manuscript writing. LYL, GY, WT, and LLL: experiment and data collection. JG and JW: data analysis. All authors contributed to the article and approved the submitted version.

Corresponding authors

Correspondence to Licai Zhang or Jiangang Song.

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The authors declare no competing interests.

Ethical approval

All experiments were approved by the Committee for the Ethical Use of Laboratory Animals at Shanghai University of Traditional Chinese Medicine and conducted in accordance with the guidelines of the International Association for the Study of Pain (IASP).

Conflicts of interest

Wei Song declares that he/she has no conflict of interest.

Yue Yong declares that he/she has no conflict of interest.

Yalan Zhou declares that he/she has no conflict of interest.

Liyue Lu declares that he/she has no conflict of interest.

Guijie Yu declares that he/she has no conflict of interest.

Wei Tang declares that he/she has no conflict of interest.

Jian Wang declares that he/she has no conflict of interest.

Jun Guo declares that he/she has no conflict of interest.

Lili Li declares that he/she has no conflict of interest.

Licai Zhang declares that he/she has no conflict of interest.

Jiangang Song declares that he/she has no conflict of interest.

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Song, W., Yong, Y., Zhou, Y. et al. Activation of P2X4 receptors in midbrain cerebrospinal fluid-contacting nucleus leads to mechanical hyperalgesia in chronic constriction injury rats. Purinergic Signalling 19, 481–487 (2023). https://doi.org/10.1007/s11302-022-09911-0

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