Inhibitory effects of AG490 on H2O2-induced TRPM2-mediated Ca2+entry

Shunichi Shimizu, Ryo Yonezawa, Tamio Hagiwara, Takashi Yoshida, Nobuaki Takahashi, Satoshi Hamano, Takaharu Negoro, Takahiro Toda, Minoru Wakamori, Yasuo Mori, Masakazu Ishii

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15 Citations (Scopus)

Abstract

Transient receptor potential melastatin 2 (TRPM2) is an oxidative stress-sensitive Ca2+-permeable channel that controls Ca2+signalling. The activation of Janus kinase 2 (Jak2) by oxidative stress is implicated in the production of inflammatory mediators. We found that AG490, a Jak2 inhibitor, had an inhibitory effect on H2O2-induced TRPM2 activation. The purpose of this study was to examine the underlying mechanisms of the inhibitory effects of AG490. Activation of TRPM2 in TRPM2-expressing human embryonic kidney 293 (TRPM2/HEK) cells or the human monocytic cell line U937 was monitored by fluorescence-based Ca2+imaging and patch-clamp techniques. Treatment with AG490 almost completely blocked H2O2-induced increase in intracellular Ca2+in TRPM2/HEK and U937 cells. In the patch-clamp study, AG490 inhibited the H2O2-evoked inward current but not the ADP-ribose-induced inward current in TRPM2/HEK cells. In contrast, Jak inhibitor 1 (pyridone 6) and staurosporine, both of which inhibit Jak2, had no effect on H2O2-induced increase in intracellular Ca2+. Moreover, AG490 decreased intracellular reactive oxygen species level, which was measured by using a hydroperoxide-sensitive fluorescent dye, on incubation with H2O2. In the cell-free assay system, AG490 scavenged hydroxyl radicals but not H2O2. These findings indicate that AG490 significantly reduces H2O2-induced TRPM2 activation, presumably by scavenging hydroxyl radicals rather than Jak2-dependent mechanisms. Although transient receptor potential ankyrin 1 (TRPA1) channel is also activated by H2O2, the H2O2-induced Ca2+entry through TRPA1 was only slightly delayed by AG490. This validates the potential use of AG490, as one of the materials for characterizing the role of TRPM2 channels in pathological models.

Original languageEnglish
Pages (from-to)22-30
Number of pages9
JournalEuropean Journal of Pharmacology
Volume742
DOIs
Publication statusPublished - 2014 Nov 5

Keywords

  • AG490
  • Janus kinase 2
  • Oxidative stress
  • TRPA1
  • TRPM2

ASJC Scopus subject areas

  • Pharmacology

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    Shimizu, S., Yonezawa, R., Hagiwara, T., Yoshida, T., Takahashi, N., Hamano, S., Negoro, T., Toda, T., Wakamori, M., Mori, Y., & Ishii, M. (2014). Inhibitory effects of AG490 on H2O2-induced TRPM2-mediated Ca2+entry. European Journal of Pharmacology, 742, 22-30. https://doi.org/10.1016/j.ejphar.2014.08.023