PMID- 27433342 OWN - NLM STAT- PubMed-not-MEDLINE DCOM- 20160719 LR - 20231111 IS - 2052-1707 (Print) IS - 2052-1707 (Electronic) IS - 2052-1707 (Linking) VI - 4 IP - 3 DP - 2016 Jun TI - Diclofenac, a nonsteroidal anti-inflammatory drug, is an antagonist of human TRPM3 isoforms. PG - e00232 LID - 10.1002/prp2.232 [doi] LID - e00232 AB - The effects of diclofenac (Dic), an acetic acid derivative-type nonsteroidal anti-inflammatory drug, were examined on the function of transient receptor potential (TRP) melastatin (TRPM) 3 (TRPM3) in human embryonic kidney 293 cell-line (HEK293) cells with recombinant human TRPM3 isoforms (TRPM31325, TRPM3-3, TRPM3-9, and TRPM3-S) and in a neuroblastoma cell line human neuroblastoma IMR-32 cells (IMR-32 cells) derived from human peripheral neurons. TRPM3 responses evoked by pregnenolone sulfate (PregS) were effectively inhibited by Dic in a concentration-dependent manner in Ca(2+) measurement and electrophysiological assays. The apparent IC 50 for PregS-induced Ca(2+) response of TRPM31325, TRPM3-3, and TRPM3-9 was calculated to be 18.8, 42.5, and 7.1 mumol/L, respectively. The TRPM3-dependent Ca(2+) responses evoked by nifedipine, another TRPM3 agonist, were also significantly inhibited by Dic. In contrast, aceclofenac, an acetoxymethyl analog of Dic, had no effects on PregS-induced TRPM3 responses. Constitutive channel activity of TRPM3-S without TRPM3 agonists was substantially inhibited by Dic, ruling out the possibility of interaction of Dic against TRPM3 agonists to the channel binding sites. Moreover, Dic reversibly inhibited TRPM3 single-channel activity recorded in excised outside-out patches without affecting the channel conductance. In differentiated neuronal IMR-32 cells with endogenous TRPM3, Dic inhibited PregS-evoked Ca(2+) responses with an apparent IC 50 of 17.1 mumol/L. Taken together, our findings demonstrate that Dic inhibits human TRPM3 without interacting with the channel pore. FAU - Suzuki, Hiroka AU - Suzuki H AD - Laboratory of Cellular Pharmacology School of Pharmacy Aichi-Gakuin University 1-100 Kusumoto Chikusa, Nagoya 464-8650 Japan. FAU - Sasaki, Eiji AU - Sasaki E AD - Laboratory of Cellular Pharmacology School of Pharmacy Aichi-Gakuin University 1-100 Kusumoto Chikusa, Nagoya 464-8650 Japan. FAU - Nakagawa, Ayumi AU - Nakagawa A AD - Laboratory of Cellular Pharmacology School of Pharmacy Aichi-Gakuin University 1-100 Kusumoto Chikusa, Nagoya 464-8650 Japan. FAU - Muraki, Yukiko AU - Muraki Y AD - Laboratory of Cellular Pharmacology School of Pharmacy Aichi-Gakuin University 1-100 Kusumoto Chikusa, Nagoya 464-8650 Japan. FAU - Hatano, Noriyuki AU - Hatano N AD - Laboratory of Cellular Pharmacology School of Pharmacy Aichi-Gakuin University 1-100 Kusumoto Chikusa, Nagoya 464-8650 Japan. FAU - Muraki, Katsuhiko AU - Muraki K AUID- ORCID: 0000-0002-4779-6494 AD - Laboratory of Cellular Pharmacology School of Pharmacy Aichi-Gakuin University 1-100 Kusumoto Chikusa, Nagoya 464-8650 Japan. LA - eng PT - Journal Article DEP - 20160407 PL - United States TA - Pharmacol Res Perspect JT - Pharmacology research & perspectives JID - 101626369 PMC - PMC4876142 OTO - NOTNLM OT - Diclofenac OT - TRPM3 OT - neuroblastoma OT - pain EDAT- 2016/07/20 06:00 MHDA- 2016/07/20 06:01 PMCR- 2016/04/07 CRDT- 2016/07/20 06:00 PHST- 2016/01/28 00:00 [received] PHST- 2016/03/04 00:00 [revised] PHST- 2016/03/07 00:00 [accepted] PHST- 2016/07/20 06:00 [entrez] PHST- 2016/07/20 06:00 [pubmed] PHST- 2016/07/20 06:01 [medline] PHST- 2016/04/07 00:00 [pmc-release] AID - PRP2232 [pii] AID - 10.1002/prp2.232 [doi] PST - epublish SO - Pharmacol Res Perspect. 2016 Apr 7;4(3):e00232. doi: 10.1002/prp2.232. eCollection 2016 Jun.