Simultaneous measurement of superoxide generation and intracellular Ca2+ concentration reveals the effect of extracellular Ca2+ on rapid and transient contents of superoxide generation in differentiated THP-1 cells

Biochemical and Biophysical Research Communications
Kaname IshibashiMitsuo Hiramatsu

Abstract

We invented a simultaneous measuring instrument of fluorescence and chemiluminescence, realizing the analysis of chronological correlation between change in intracellular Ca2+ concentration ([Ca2+]i) and superoxide generation. A human monocytic cell line, THP-1, differentiated to be neutrophil-like cells generated superoxide with increase in intracellular Ca2+ concentration when stimulated with formyl-methionyl-leucyl-phenylalanine (fMLP) whereas PMA, phorbol ester-stimulated superoxide response occurred without change in [Ca2+]i. The cells treated with TMB-8, an intracellular Ca2+ antagonist, generated superoxide rapidly as well as transiently with transient [Ca2+]i elevation after stimulation with fMLP, whereas EGTA-treated cells generated superoxide slowly as well as persistently with transient [Ca2+]i elevation after the stimulation. These results suggest that the rapid and transient contents of superoxide generation are specific for Ca2+ influx from the extracellular domain. Verapamil, voltage-dependent Ca2+ channel blocker, dose-dependently inhibited fMLP-stimulated extracellular Ca2+ influx and superoxide generation without affecting PMA-stimulated superoxide generation. Other channel blockers tested, nifedipine and dilt...Continue Reading

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Citations

May 29, 2007·Infection and Immunity·Jens Martin HerrmannElizabeth R Simons
Feb 6, 2010·Biochemical Pharmacology·Tomoaki IshiharaTohru Mizushima
Nov 7, 2006·Archives of Biochemistry and Biophysics·Roberto StifaneseEdon Melloni
Sep 13, 2014·Journal of Thrombosis and Thrombolysis·Katsuyasu SaigoShion Imoto
Aug 9, 2016·Journal of Clinical Biochemistry and Nutrition·Kimiko KazumuraHiromi Takano-Ohmuro
Jul 23, 2013·The Journal of Immunology : Official Journal of the American Association of Immunologists·Soad M SalehFutwan A Al-Mohanna
May 15, 2013·The Journal of Immunology : Official Journal of the American Association of Immunologists·Shun-Chin YangTsong-Long Hwang

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