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Activation of TRPA1 channels by fenamate nonsteroidal anti-inflammatory drugs

  • Ion Channels, Receptors and Transporters
  • Published:
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Abstract

Transient receptor potential A1 (TRPA1) forms nonselective cation channels implicated in acute inflammatory pain and nociception. The mechanism of ligand activation of TRPA1 may involve either covalent modification of cysteine residues or conventional reversible ligand–receptor interactions. For certain electrophilic prostaglandins, covalent modification has been considered as the main mechanism involved in their stimulatory effect on TRPA1. Because some nonsteroidal anti-inflammatory drugs (NSAIDs) are structural analogs of prostaglandins, we examined several nonelectrophilic NSAIDs on TRPA1 activation using electrophysiological techniques and intracellular Ca2+ measurements and found that a selected group of NSAIDs can act as TRPA1 agonists. Extracellularly applied flufenamic, niflumic, and mefenamic acid, as well as flurbiprofen, ketoprofen, diclofenac, and indomethacin, rapidly activated rat TRPA1 expressed in Xenopus oocytes and human TRPA1 endogenously expressed in WI-38 fibroblasts. Similarly, the NSAID ligands activated human TRPA1 inducibly expressed in HEK293 cells, but the responses were absent in uninduced and parental HEK293 cells. The response to fenamate agonists was blocked by TRPA1 antagonists, AP-18, HC-030031, and ruthenium red. At subsaturating concentrations, the fenamate NSAIDs also potentiate the activation of TRPA1 by allyl isothiocyanate, cinnamaldehyde, and cold, demonstrating positive synergistic interactions with other well-characterized TRPA1 activators. Importantly, among several thermosensitive TRP channels, the stimulatory effect is specific to TRPA1 because flufenamic acid inhibited TRPV1, TRPV3, and TRPM8. We conclude that fenamate NSAIDs are a novel class of potent and reversible direct agonists of TRPA1. This selective group of TRPA1-stimulating NSAIDs should provide a structural basis for developing novel ligands that noncovalently interact with TRPA1 channels.

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Abbreviations

2APB:

2-Aminoethoxydiphenyl borate

AITC:

Allyl isothiocyanate

CA:

Cinnamaldehyde

[Ca2+]i :

Intracellular Ca2+ concentration

COX:

Cyclooxygenase

ECS:

Extracellular solution

FFA:

Flufenamic acid

MFA:

Mefenamic acid

NFA:

Niflumic acid

NSAIDs:

Nonsteroidal anti-inflammatory drugs

TRP:

Transient receptor potential

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Acknowledgements

We thank Drs. Ardem Patapoutian and Michael Bandell for the HEK-TRPA1 cell line, Dr. Jun Chen for helpful discussion of the project, Ms. Dina Chuang-Zhu for technical assistance. This work was supported by American Heart Association Grant-in-Aid 0755277B and US NIH grants RO1-NS042183, RO1-DK081654, R21-NS056942, and P30-NS045758.

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Correspondence to Michael X. Zhu.

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Hongzhen Hu, Jinbin Tian, and Yingmin Zhu contributed equally to this work.

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Hu, H., Tian, J., Zhu, Y. et al. Activation of TRPA1 channels by fenamate nonsteroidal anti-inflammatory drugs. Pflugers Arch - Eur J Physiol 459, 579–592 (2010). https://doi.org/10.1007/s00424-009-0749-9

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  • DOI: https://doi.org/10.1007/s00424-009-0749-9

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