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The real-time detection of acupuncture-induced extracellular ATP mobilization in acupoints and exploration of its role in acupuncture analgesia

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Abstract

Our and in vitro studies had confirmed that mechanosensitive ATP release and accumulation in acupoints was elicited by acupuncture (AP), which might be a pivotal step for triggering AP analgesia. But to date, the dynamics of extracellular ATP (eATP) in the interstitial space during AP process was poorly known, mainly due to the low temporal resolution of the current detection approach. This study attempted to capture rapid eATP signals in vivo in the process of needling, and further explored the role of this eATP mobilization in initiating AP analgesic effect. Ipsilateral 20-min needling was applied on Zusanli acupoint (ST36) of complete Freund’s adjuvant (CFA)–induced ankle arthritis rats. Pain thresholds were assessed in injured-side hindpaws. eATP in the interstitial space was microdialyzed and real-time quantified by luciferin-luciferase assay at 1-min interval with the aid of the microfluid chip. We revealed in behavioral tests that modulation of eATP levels in ST36 influenced AP analgesic effect on ankle arthritis. A transient eATP accumulation was induced by needling that started to mobilize at 4 min, climbed to the peak of 11.21 nM within 3.25 min and gradually recovered. Such AP-induced eATP mobilization was significantly impacted by ankle inflammation, needling depth, needle manipulation, and the presence of local ecto-nucleotidases. This work reveals that needling elicits a transient eATP mobilization in acupoints, which contributes to initiating AP analgesia. This study will help us better understand the peripheral mechanism of AP analgesia and guide clinicians to optimize the needle manipulations to improve AP efficacy.

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Availability of data and material

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

Abbreviations

Ado:

Adenosine

ARL67156:

6-N,N-Diethyl-β-γ-dibromomethylene-d-adenosine-5′-triphosphate

CFA:

Complete Freund’s adjuvant

eATP:

Extracellular ATP

[eATP]:

Concentration of extracellular ATP

HBSS:

Hank’s balanced salt solution

HPLC:

High-performance liquid chromatography

L-L:

Luciferin-luciferase

Nt5e:

Ecto-5′-nucleotidase

NTPDases:

Nucleoside triphosphate diphosphohydrolases

PAP:

Prostatic acid phosphatase

P2 receptors:

Purinergic receptors

PDMS:

Polydimethylsiloxane

PWL:

Paw withdraw latency

PWT:

Paw withdraw threshold

TNAP:

Tissue nonspecific alkaline phosphatase

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Acknowledgements

We like to thank Prof. Ryszard Grygorczyk (Université de Montréal, Montreal, Canada) and Prof. Wolfgang Schwarz (Goethe-University, Frankfurt, Germany) for advice on microfluid chip.

Funding

This study received funding from the National Natural Science Foundation of China (Grant No. 81574076 to L-N W, 82174488 to G-H D, and 82105005 to D S), Budget Research Project of Shanghai Education Commission (Grant No. 2021LK098), Shanghai Key Laboratory of Acupuncture Mechanism and Acupoint Function (Grant No. 21DZ2271800 to G-H D), and Postgraduate Innovation Training Project of Shanghai University of Traditional Chinese Medicine (Grant No. Y2021025 to Y-J L).

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Authors and Affiliations

  1. School of Acupuncture-Moxibustion and Tuina, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai, 201203, China

    Wei-Min Zuo, Yu-Jia Li, Kai-Yu Cui, Dan Shen, Ya-Wen Zheng, Xue-Yong Shen & Li-Na Wang

  2. HuBei Provincial Hospital of Traditional Chinese Medicine, 4 Huayuanshan Road, Wuhan, 430060, Hubei, China

    Wei-Min Zuo

  3. School of Traditional Chinese Medicine, Naval Medical University, 800 Xiangyin Road, Shanghai, 200433, China

    Dan Shen

  4. Shanghai Key Laboratory of Acupuncture Mechanism and Acupoint Function (14DZ2260500), Fudan University, 220 Handan Road, Shanghai, 201433, China

    Di Zhang, Meng Huang, Li-Na Wang & Guang-Hong Ding

  5. Department of Aeronautics and Astronautics, Fudan University, 220 Handan Road, Shanghai, 200433, China

    Di Zhang, Yong Wu & Guang-Hong Ding

  6. Shanghai Research Center for Acupuncture and Meridians, Shanghai, 201203, China

    Xue-Yong Shen

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  1. Wei-Min Zuo
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  2. Yu-Jia Li
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  3. Kai-Yu Cui
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  4. Dan Shen
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  5. Di Zhang
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  10. Li-Na Wang
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  11. Guang-Hong Ding
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Contributions

Conceptualization by Li-Na Wang, Guang-Hong Ding, and Xue-Yong Shen; data collection and figure formation by Wei-Min Zuo, Yu-Jia Li, Kai-Yu Cui, Dan Shen, and Ya-Wen Zheng; data analysis by Wei-Min Zuo, Yu-Jia Li; figure formation by Wei-Min Zuo, Yu-Jia Li, Kai-Yu Cui, Dan Shen; methodology by Wei-Min Zuo, Meng Huang, and Yong Wu; data interpretation by Li-Na Wang; first manuscript writing by Wei-Min Zuo; manuscript revision by Li-Na Wang; funding acquisition by Li-Na Wang and Di Zhang. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Li-Na Wang or Guang-Hong Ding.

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All experimental protocols were approved by the Animal Care Committee of Shanghai University of Traditional Chinese Medicine (Shanghai, China) (No SZY201807008).

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Zuo, WM., Li, YJ., Cui, KY. et al. The real-time detection of acupuncture-induced extracellular ATP mobilization in acupoints and exploration of its role in acupuncture analgesia. Purinergic Signalling 19, 69–85 (2023). https://doi.org/10.1007/s11302-021-09833-3

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