Relationship between calcium release and NADPH oxidase inhibition in human neutrophils
Michael D Salmon1, Jatinder Ahluwalia
1Leukocyte and Ion Channel Research Laboratory, School of Health and Biosciences, University of East London, London, UK.
Abstract:
The aim of this study was to investigate the possible relationship between NADPH oxidase activity and changes in cytosolic Ca(2+) in response to different agonists. Treatment of neutrophils with leukotriene B4 (LTB(4)) demonstrated characteristic changes to cytoslic Ca(2+) yielding an EC(50) of 4nM. The pA(2) values for the specific LTB(4) receptor (BLT) antagonists, U-75302 and LY-255283 were 6.32 and 6.38, respectively. Similarly, neutrophils treated with N-formyl-l-methionyl-l-leucyl-l-phenylalanine (FMLP) and platelet activating factor (PAF) exhibited changes in cytoslic Ca(2+) in a dose dependant manner with pD(2) values of 9.0 and 9.9, respectively. The phorbol ester PMA prevented elevations in cytosolic Ca(2+) in response to LTB(4), FMLP and PAF with IC(50) values of 5.88, 1.44 and 5.71nM, respectively. In addition, potent NADPH oxidase inhibitors apocynin and diphenyleneiodonium (DPI) inhibited FMLP mediated cytosolic Ca(2+) release. These results demonstrate that inhibition of the NADPH oxidase suppresses cytosolic Ca(2+) release in FMLP activated human neutrophils.
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