Direct non-genomic effect of steroid hormones on superoxide anion generation in the bone resorbing osteoclasts.

We have investigated the possible acute effect of steroid hormones, including 1alpha,25 dihydroxycholecalciferol (1alpha,25(OH)2D3) and estradiol, on the generation of superoxide anion (O2*-) in bone resorbing osteoclasts. Evidence is presented demonstrating acute non-genomic stimulatory action of 1alpha,25(OH)2D3 on the production of free radicals by rat osteoclasts cultured on calcified matrix. The increase in O2*- production was observed in the range of 6-10 s (n = 5) following exposure of enriched osteoclasts to 1alpha,25(OH)2D3 and was found to be transient with the peak response being in the range of 5-45 s (n = 5). The decline in the transient was much slower than the elevation, time for the decay being in the range 1-5 min (n = 5) and remained above the levels present prior to the addition. The exposure of the osteoclast to dexamethasone was found to have no effect on O2*- generation, whilst estradiol was found to be inhibitory. The mode of stimulation and the kinetics of the transients of O2*- in the bone resorbing osteoclasts produced by 1alpha,25(OH)2D3 were similar to that of parathyroid hormone (PTH) and pertussis. The exposure of the bone resorbing osteoclasts to cholera toxin was found to have no effect, suggesting that the stimulatory action is unlikely to be mediated via cAMP elevation. The importance of these observations is discussed in the context of calcium homeostasis and bone physiology.