1. Rat peritoneal macrophages stimulated with phorbol 12-myristate 13-acetate (PMA) (40 nM) show an increase in the rate of oxygen consumption (measured with an O2 electrode) and the production of superoxide (measured by cytochrome c reduction), which are both dependent on the presence of exogenous glucose. There is a 1:1 correlation between the oxygen consumed and the superoxide produced over a range of glucose concentrations (0-10 mM). 2. Preincubation of macrophages with dexamethasone (1 microM) for 3 h significantly decreased the Vmax. for PMA-induced glucose-dependent oxygen consumption (P < 0.001) and glucose-dependent superoxide production (P < 0.001). However, dexamethasone did not significantly change the Km for glucose in either PMA-induced oxygen consumption or superoxide production. Dexamethasone is therefore a non-competitive inhibitor of PMA-stimulated glucose-dependent oxygen consumption (Ki = 0.83 +/- 0.09 microM) and superoxide generation (Ki = 0.87 +/- 0.09 microM). 3. The PMA-induced rate of oxygen consumption by macrophages was decreased at oxygen concentrations below approx. 15 microM. The Km of oxygen for PMA-induced oxygen consumption was 1.28 +/- 0.13 microM (n = 12), and this was not significantly different in the presence of dexamethasone; Km = 1.61 +/- 0.31 microM (n = 12). It is therefore concluded that in vivo macrophage superoxide production is not limited by external oxygen or glucose concentrations, even in hypoxic joints.
- © 1993 The Biochemical Society, London