Inositol 1,4,5-Trisphosphate Receptor-Mediated Initial Ca2+ Mobilization Constitutes a Triggering Signal for Hydrogen Peroxide-Induced Apoptosis in INS-1 β-Cells
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概要
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Reactive oxygen species, including hydrogen peroxide (H2O2), are known to induce β-cell apoptosis. The present study investigated the role of Ca2+ in H2O2-induced apoptosis of the β-cell line INS-1. Annexin V assay with flow cytometry and DNA ladder assay demonstrated that treatment of INS-1 cells with 100 μM H2O2 for 18 h significantly increased apoptotic cells. A comparable level of apoptosis was also observed after 18 h when the cells were treated with 100 μM H2O2 only for initial 30 min. The H2O2-induced apoptosis was abolished by 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetra(acetoxymethyl)ester (BAPTA/AM), a chelator of intracellular Ca2+, by 2-aminoethoxydiphenylborate (2-APB), a blocker of inositol 1,4,5-trisphosphate (IP3) receptors and cation channels, and by xestospongin D, a blocker of IP3 receptors, and was partially blocked by SKF-96365, a non-selective cation channel blocker. However, nicardipine, an L-type voltage-dependent Ca2+ channel blocker, or N-(p-amylcinnamoyl)anthranilic acid (ACA), a TRPM2 blocker, had little effect on the apoptosis. The inhibitory effect of BAPTA/AM or 2-APB on the H2O2-induced apoptosis was largely attenuated when the drug was added 30 min or 1 h after start of the treatment with H2O2. These results suggest that the initial intracellular Ca2+ elevation induced by H2O2, which is mediated via IP3 receptors and store-operated cation channels, plays an obligatory role in the induction of β-cell apoptosis.
著者
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Takada Masahiro
Department Of Astronomy
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ISHIKAWA Tomohisa
Department of Neuropsychiatry, Ehime University Medical School
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Noguchi Akiko
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, University of Shizuoka
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Sayama Yurie
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, University of Shizuoka
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Kurohane Kaneko
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, University of Shizuoka
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Ishikawa Tomohisa
Department Of Cellular And Molecular Pharmacology Graduate School Of Pharmaceutical Sciences Univers
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Ishikawa Tomohisa
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, University of Shizuoka
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Takada Masahiro
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, University of Shizuoka
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