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Electroacupuncture inhibits visceral pain via adenosine receptors in mice with inflammatory bowel disease

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Abstract

To investigate the involvement of peripheral adenosine receptors in the effect of electroacupuncture (EA) on visceral pain in mice with inflammatory bowel disease (IBD). 2,4,6-Trinitrobenzene sulfonic acid (TNBS) was used to induce the visceral pain model. EA (1 mA, 2 Hz, 30 min) treatment was applied to bilateral acupoints “Dachangshu” (BL25) 1 day after TNBS injection once daily for 7 consecutive days. Von Frey filaments were used to measure the mechanical pain threshold. Western blot was used to detect the protein expression levels of adenosine 1 receptor (A1R), adenosine 2a receptor (A2aR), adenosine 2b receptor (A2bR), adenosine 3 receptor (A3R), substance P (SP), and interleukin 1 beta (IL-1β) in colon tissue. EA significantly ameliorated the disease-related indices and reduced the expression of SP and IL-1β in the colon tissues of mice with IBD. EA increased the expression of A1R, A2aR, and A3R and decreased the expression of A2bR in the colon tissue. Furthermore, the administration of adenosine receptor antagonists influenced the effect of EA. EA can inhibit the expression of the inflammatory factors SP and IL-1β by regulating peripheral A1, A2a, A2b, and A3 receptors, thus inhibiting visceral pain in IBD mice.

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Funding

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by a grant from the National Natural Science Foundation of China (81674057) and the major project of National Natural Science Foundation of Hubei province (No. 2015CFA094).

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Authors and Affiliations

  1. Department of Neurobiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, 430030, China

    Tengfei Hou, Hongchun Xiang, Hongping Li, He Zhu, Lixue Lin, Xuefei Hu, Hong Zhang & Man Li

  2. Institute of Integrated Traditional Chinese and Western Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Lingling Yu

  3. Department of Acupuncture, Wuhan Integrated TCM & Western Medicine Hospital, 215 Zhongshan Avenue, Wuhan, 430022, China

    Wen Su

  4. Department of Central Laboratory, Affiliated Hospital of Jiangsu University, Zhenjiang, 212000, China

    Yang Shu

  5. Department of Physiology, Basic Medical College of Nanchang University, Nanchang, 330006, China

    Shangdong Liang

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  1. Tengfei Hou
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All experimental procedures were approved by the Animal Care Committee at Huazhong University of Science and Technology and conformed to the ethical guidelines of the International Association for the Study of Pain (IASP). All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Supplementary Figure 1

Experimental design timeline. Abbreviations: EA, electroacupuncture; CLM, colon length measurement; WB, Western blotting. a Design timeline of experiment 1. The mechanical thresholds, body weight and diarrhoea scores were tested for 3 days before TNBS/vehicle injection and the average of which was calculated as the baseline, behaviour tests (mechanical thresholds, body weight and diarrhoea scores) were observed once daily for 7 consecutive days 1 day after vehicle/TNBS injection. In the TNBS+EA/ sham EA group, EA/sham EA was applied once daily for 7 consecutive days 1 day after TNBS injection. Behaviour tests (mechanical thresholds, body weight and diarrhoea scores) were observed after EA/sham EA treatment every day. After the last time of behaviour tests, mice were deeply anaesthetised and their descending colon tissues were removed for colon length measurement and Western blotting. b Design timeline of experiment 2. The mechanical thresholds, body weight and diarrhoea scores were tested for 3 days before TNBS/vehicle injection and the average of which was calculated as the baseline, behaviour tests (mechanical thresholds, body weight and diarrhoea scores) were observed once daily for 7 consecutive days 1 day after vehicle/TNBS injection.In the TNBS+EA/ sham EA group, EA/sham EA was applied once daily for 7 consecutive days 1 day after TNBS injection. In the TNBS+EA + DPCPX/ZM241385/PSB603/MRS3777 group, 4 kinds of adenosine receptor antagonists were injected 30 min before EA treatment every day, respectively. Behaviour tests were observed after EA treatment every day. After the last time of behaviour tests, mice were deeply anaesthetised and their descending colon tissues were removed for colon length measurement and Western blotting. ○, EA treatment; ●, sham EA treatment; △, EA treatment + A1R antagonist; ▲, EA treatment + A2aR antagonist; ◊, EA treatment + A2bR antagonist; ♦, EA treatment + A3R antagonist. (PNG 502 kb)

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Hou, T., Xiang, H., Yu, L. et al. Electroacupuncture inhibits visceral pain via adenosine receptors in mice with inflammatory bowel disease. Purinergic Signalling 15, 193–204 (2019). https://doi.org/10.1007/s11302-019-09655-4

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