Quantitative Fluorescence Measurement of Cardiac Na+/Ca2+ Exchanger Inhibition by Kinetic Analysis in Stably Transfected HEK293 Cells
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概要
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We developed a method to quantitatively evaluate the potency of Na+/Ca2+ exchanger (NCX) inhibitors with fluorescence microscopy in NCX1-transfected HEK 293 cells. The reverse mode and forward mode NCX activities were measured as the ascending slope of the early phase increase in cytoplasmic Ca2+ concentration after change to low Na+ extracellular solution and the descending rate (inverse of the exponential time constant) on return to normal solution, respectively. Both modes of NCX were inhibited by SEA0400 (2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline) and KB-R7943 (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea methanesulfonate), and the concentration-inhibition relationships for both inhibitors were in good agreement with those previously reported in voltage clamped cardiomyocytes.
著者
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Kawanishi Toru
Division Of Biological Chemistry And Biologicals National Institute Of Health Sciences
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Shigenobu Koki
Department Of Chemical Pharmacology Faculty Of Pharmaceutical Sciences The University Of Tokyo
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Namekata Iyuki
Department Of Pharmacology Faculty Of Pharmaceutical Sciences Toho University
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Tanaka Hikaru
Department of Endocrinology and Metabolism, Hirosaki University
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Iida-Tanaka Naoko
Department of Biochemistry, Teikyo University School of Medicine
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Namekata Iyuki
Department of Pharmacology, Toho University Faculty of Pharmaceutical Sciences, Japan
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Kawanishi Toru
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Japan
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Iida-Tanaka Naoko
Department of Pharmacology, Toho University Faculty of Pharmaceutical Sciences, Japan
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