We examined the digitonin-stimulated release of superoxide by alveolar macrophages (AMs) from young rats exposed to greater than 95% oxygen for 24-168 h. AMS were obtained by lung lavages, and the release of superoxide stimulated by digitonin was measured by using cytochrome C reduction. The total cell count of lung lavages decreased at 24 and 168 h of oxygen exposure (p less than 0.05 for both). The contamination of polymorphonuclear cells (PMNs) was less than 1% up to 120 h of oxygen exposure, and at 168 h PMNs increased to 5% of total cells in lung lavages. The viability of AMs was greater than 95% up to 72 h and then decreased to 90% at 120 h of oxygen exposure and 87% at 168 h. Digitonin-stimulated superoxide release by AMs recovered from lung lavages in rats exposed to hyperoxia showed a slight increase during the first 48 h. However, oxygen exposure for 72 h or more caused a significant decrease of the stimulated superoxide release of AMs compared to AMs from control rats. This decline in stimulated superoxide release of AMs resulting from hyperoxia was not prevented by vitamin E treatment.