Tags

Type your tag names separated by a space and hit enter

The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor.
Am J Physiol Cell Physiol. 2014 Aug 15; 307(4):C384-94.AJ

Abstract

Transient receptor potential ankyrin 1 (TRPA1) is a Ca(2+)-permeable nonselective cation channel expressed in neuronal and nonneuronal cells and plays an important role in acute and inflammatory pain. Here, we show that an NADPH oxidase (NOX) inhibitor, diphenyleneiodonium (DPI), functions as a TRPA1 activator in human embryonic kidney cells expressing human TRPA1 (HEK-TRPA1) and in human fibroblast-like synoviocytes. Application of DPI at 0.03-10 μM induced a Ca(2+) response in HEK-TRPA1 cells in a concentration-dependent manner. The Ca(2+) response was effectively blocked by a selective TRPA1 antagonist, HC-030031 (HC). In contrast, DPI had no effect on HEK cells expressing TRPV1-V4 or TRPM8. Four other NOX inhibitors, apocynin (APO), VAS2870 (VAS), plumbagin, and 2-acetylphenothiazine, also induced a Ca(2+) response in HEK-TRPA1 cells, which was inhibited by pretreatment with HC. In the presence of 5 mM glutathione, the Ca(2+) response to DPI was effectively reduced. Moreover, mutation of cysteine 621 in TRPA1 substantially inhibited the DPI-induced Ca(2+) response, while it did not inhibit the APO- and VAS-induced responses. The channel activity was induced by DPI in excised membrane patches with both outside-out and inside-out configurations. Internal application of neomycin significantly inhibited the DPI-induced inward currents. In inflammatory synoviocytes with TRPA1, DPI evoked a Ca(2+) response that was sensitive to HC. In mice, intraplantar injection of DPI caused a pain-related response which was inhibited by preadministration with HC. Taken together, our findings demonstrate that DPI and other NOX inhibitors activate human TRPA1 without mediating NOX.

Authors+Show Affiliations

Laboratory of Cellular Pharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan;Laboratory of Cellular Pharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan;Laboratory of Cellular Pharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan;Department of Drug Metabolism and Disposition, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan;Laboratory of Neuropharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan; and.Department of Drug Metabolism and Disposition, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan;Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.Laboratory of Neuropharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan; and.Laboratory of Neuropharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan; and.Laboratory of Cellular Pharmacology, School of Pharmacy, Aichi-Gakuin University, Nagoya, Japan; kmuraki@dpc.agu.ac.jp.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24944203

Citation

Suzuki, Hiroka, et al. "The NADPH Oxidase Inhibitor Diphenyleneiodonium Activates the Human TRPA1 Nociceptor." American Journal of Physiology. Cell Physiology, vol. 307, no. 4, 2014, pp. C384-94.
Suzuki H, Hatano N, Muraki Y, et al. The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor. Am J Physiol Cell Physiol. 2014;307(4):C384-94.
Suzuki, H., Hatano, N., Muraki, Y., Itoh, Y., Kimura, S., Hayashi, H., Onozaki, K., Ohi, Y., Haji, A., & Muraki, K. (2014). The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor. American Journal of Physiology. Cell Physiology, 307(4), C384-94. https://doi.org/10.1152/ajpcell.00182.2013
Suzuki H, et al. The NADPH Oxidase Inhibitor Diphenyleneiodonium Activates the Human TRPA1 Nociceptor. Am J Physiol Cell Physiol. 2014 Aug 15;307(4):C384-94. PubMed PMID: 24944203.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The NADPH oxidase inhibitor diphenyleneiodonium activates the human TRPA1 nociceptor. AU - Suzuki,Hiroka, AU - Hatano,Noriyuki, AU - Muraki,Yukiko, AU - Itoh,Yuka, AU - Kimura,Satoko, AU - Hayashi,Hidetoshi, AU - Onozaki,Kikuo, AU - Ohi,Yoshiaki, AU - Haji,Akira, AU - Muraki,Katsuhiko, Y1 - 2014/06/18/ PY - 2014/6/20/entrez PY - 2014/6/20/pubmed PY - 2014/10/10/medline KW - NADPH oxidase inhibitors KW - calcium channel KW - transient receptor potential SP - C384 EP - 94 JF - American journal of physiology. Cell physiology JO - Am J Physiol Cell Physiol VL - 307 IS - 4 N2 - Transient receptor potential ankyrin 1 (TRPA1) is a Ca(2+)-permeable nonselective cation channel expressed in neuronal and nonneuronal cells and plays an important role in acute and inflammatory pain. Here, we show that an NADPH oxidase (NOX) inhibitor, diphenyleneiodonium (DPI), functions as a TRPA1 activator in human embryonic kidney cells expressing human TRPA1 (HEK-TRPA1) and in human fibroblast-like synoviocytes. Application of DPI at 0.03-10 μM induced a Ca(2+) response in HEK-TRPA1 cells in a concentration-dependent manner. The Ca(2+) response was effectively blocked by a selective TRPA1 antagonist, HC-030031 (HC). In contrast, DPI had no effect on HEK cells expressing TRPV1-V4 or TRPM8. Four other NOX inhibitors, apocynin (APO), VAS2870 (VAS), plumbagin, and 2-acetylphenothiazine, also induced a Ca(2+) response in HEK-TRPA1 cells, which was inhibited by pretreatment with HC. In the presence of 5 mM glutathione, the Ca(2+) response to DPI was effectively reduced. Moreover, mutation of cysteine 621 in TRPA1 substantially inhibited the DPI-induced Ca(2+) response, while it did not inhibit the APO- and VAS-induced responses. The channel activity was induced by DPI in excised membrane patches with both outside-out and inside-out configurations. Internal application of neomycin significantly inhibited the DPI-induced inward currents. In inflammatory synoviocytes with TRPA1, DPI evoked a Ca(2+) response that was sensitive to HC. In mice, intraplantar injection of DPI caused a pain-related response which was inhibited by preadministration with HC. Taken together, our findings demonstrate that DPI and other NOX inhibitors activate human TRPA1 without mediating NOX. SN - 1522-1563 UR - https://www.unboundmedicine.com/medline/citation/24944203/The_NADPH_oxidase_inhibitor_diphenyleneiodonium_activates_the_human_TRPA1_nociceptor_ L2 - https://journals.physiology.org/doi/10.1152/ajpcell.00182.2013?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -