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Electroacupuncture Alleviates Pain by Suppressing P2Y12R-Dependent Microglial Activation in Monoarthritic Rats

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Abstract

Electroacupuncture (EA) effectively improves arthritis-induced hyperalgesia and allodynia by repressing spinal microglial activation, which plays a crucial role in pain hypersensitivity following tissue inflammation. However, the mechanism by which EA suppresses spinal microglial activation in monoarthritis (MA) remains unclear. In the present study, a rat model of MA was established through unilateral ankle intra-articular injection of complete Freund’s adjuvant (CFA). The relationship among P2Y12 receptor (P2Y12R) expression, spinal microglial activation, and EA analgesia was investigated using quantitative real-time PCR (qRT‒PCR), western blotting, immunofluorescence (IF), and behavioral testing. The results found that EA treatment at the ipsilateral “Huantiao” (GB30) and “Yanglingquan” (GB34) acupoints markedly attenuated pain and spinal microglia M1 polarization in MA rats. In particular, P2Y12R expression was significantly increased at the mRNA and protein levels in the spinal dorsal horn in MA rats, whereas EA treatment effectively repressed the MA-induced upregulation of P2Y12R. IF analysis further revealed that most P2Y12R was expressed in microglia in the spinal dorsal horn. Pharmacological inhibition of P2Y12R by its antagonist (AR-C69931MX) decreased MA-induced spinal microglial activation and subsequent proinflammatory cytokine production. Consequently, AR-C69931MX significantly intensified the anti-pain hypersensitive function of EA in MA rats. Taken together, these results demonstrate that EA alleviates MA-induced pain by suppressing P2Y12R-dependent microglial activation.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC, 81904283), Special Project for Clinical Research in the Health Industry of the Shanghai Municipal Health Commission (No.20194Y0307), and Shanghai University of Traditional Chinese Medicine project (2021LK082).

Funding

This work was supported by the National Natural Science Foundation of China (NSFC, 81904283), Special Project for Clinical Research in the Health Industry of the Shanghai Municipal Health Commission (No.20194Y0307), Shanghai University of Traditional Chinese Medicine project (2021LK082), and Shanghai Traditional Chinese Medicine "Three-year Action Plan" [ZY(2021-2023)-0211].

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  1. Jing Wang, Wei Song and Yujiao Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Anesthesiology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, No. 185, Pu An Road, Shanghai, 201203, China

    Jing Wang, Wei Song, Yujiao Zhang, Jian Wang, Yongqiang Wang, Jiangang Song & Yalan Zhou

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Contributions

Conceptualization and methodology: J.W., Y.Z. Resources, data curation and visualization: W.S., J.W., Y.W. Writing- original draft preparation: J.S. Supervision, writing- reviewing and editing: Y.Z.

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Correspondence to Jiangang Song or Yalan Zhou.

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Animal experiments were performed with the approval of the Animal Ethics Committee of the Shanghai University of Traditional Chinese Medicine (201801080001), in line with the ARRIVE guidelines to minimize animal suffering.

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Wang, J., Song, W., Zhang, Y. et al. Electroacupuncture Alleviates Pain by Suppressing P2Y12R-Dependent Microglial Activation in Monoarthritic Rats. Neurochem Res 49, 1268–1277 (2024). https://doi.org/10.1007/s11064-024-04114-y

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