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 4 nM. 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 5.71 nM, 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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