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Spinal cord astrocyte P2X7Rs mediate the inhibitory effect of electroacupuncture on visceral hypersensitivity of rat with irritable bowel syndrome

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Abstract

This study explored the role of P2X7 receptors in spinal cord astrocytes in the electroacupuncture-induced inhibition of visceral hypersensitivity (VH) in rats with irritable bowel syndrome (IBS). Visceral hypersensitivity of IBS was intracolonically induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). Visceromotor responses to colorectal distension (CRD-20,40,60,80 mmHg) and abdominal withdrawal reflex scoring (AWRs) were recorded after electroacupuncture at bilateral Zusanli (ST36) and Sanyinjiao (SP6) acupoints to evaluate the analgesic effect of electroacupuncture on visceral pain in rats with IBS. Fluorocitric acid (FCA), an astrocyte activity inhibitor, was injected intrathecally before electroacupuncture intervention and AWRs were recorded. Western blot and real-time qPCR were used to detect the expression of NMDA and P2X7 receptor to observe the regulation effect of electroacupuncture on NMDA receptor in the spinal cord of rats with visceral hypersensitivity. Intrathecal injection of P2X7 agonist or antagonist was administered before electroacupuncture treatment. To observe the effect of P2X7 receptor in spinal astrocytes on the inhibition of visceral hyperalgesia by electroacupuncture, the changes of AWR score, NMDA receptor in the spinal cord, and GFAP expression in astrocytes were detected. Inflammation of the colon had basically subsided at day 21 post-TNBS; persistent visceral hypersensitivity could be suppressed by electroacupuncture. This analgesic effect could be inhibited by FCA. The analgesic effect, downregulation of NMDA receptor NR1 subunit, and P2X7 protein of electroacupuncture were all reversed by FCA. P2X7 receptor antagonist A740003 can cooperate with EA to carry out analgesic effect in rats with visceral pain and downregulate the expression of NR1, NR2B, and GFAP in spinal dorsal horn. However, the P2X7 receptor agonist BzATP could partially reverse the analgesic effect of EA, inhibiting the downregulatory effect of EA on the expression of NR1, NR2B, and GFAP. These results indicate that EA may downregulate the expression of the NMDA receptor by inhibiting the P2X7 receptor in the spinal cord, thereby inhibiting spinal cord sensitization in IBS rats with visceral pain, in which astrocytes are an important medium.

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Funding

This study was supported by the Health Industry Clinical Research Project of Shanghai Municipal Health Commission (201940064), the National Natural Science Foundation of China (81873367); Three-year Action Plan Project of Shanghai Traditional Chinese Medicine Development, No. ZY(2018–2020)-CCCX-2004–01; National Traditional Chinese Medicine Leading Talents Support Program-Qihuang Scholar.

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Authors and Affiliations

  1. Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 201203, China

    Zhi-jun Weng, Shi-xiu Hu, Fang Zhang, Zhi-ying Zhang, Yun Zhou, Min Zhao, Yan Huang, Huan-gan Wu & Hui-rong Liu

  2. Fudan University Shanghai Cancer Center, Shanghai, 200032, China

    Yu-hu Xin

  3. Key Laboratory of Acupuncture-Moxibustion and Immunology, Shanghai University of Traditional Chinese Medicine, No. 650 South Wanping Road, Xuhui District, Shanghai, 201203, China

    Huan-gan Wu & Hui-rong Liu

Authors
  1. Zhi-jun Weng
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  2. Shi-xiu Hu
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  3. Fang Zhang
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  4. Zhi-ying Zhang
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  5. Yun Zhou
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  6. Min Zhao
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  7. Yan Huang
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  8. Yu-hu Xin
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  9. Huan-gan Wu
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  10. Hui-rong Liu
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Contributions

Huangan Wu and Huirong Liu designed the research; Zhijun Weng, Min Zhao, Zhiying Zhang, and Yun Zhou performed the experiments; Fang Zhang and Yan Huang collected and analyzed the data; Yuhu Xin performed literature retrieval. Zhijun Weng and Shixiu Hu wrote the manuscript; all authors reviewed the manuscript prior to its submission, and read and approved the final manuscript. Zhijun Weng and Shixiu Hu contributed equally to this study.

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Correspondence to Huan-gan Wu or Hui-rong Liu.

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All experiments were approved by the Committee for Medical Ethics and the Use of Experimental Animals at Shanghai University of Traditional Chinese Medicine.

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Weng, Zj., Hu, Sx., Zhang, F. et al. Spinal cord astrocyte P2X7Rs mediate the inhibitory effect of electroacupuncture on visceral hypersensitivity of rat with irritable bowel syndrome. Purinergic Signalling 19, 43–53 (2023). https://doi.org/10.1007/s11302-021-09830-6

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