Previous studies with lung homogenates and isolated cells have suggested oxygen cell injury results from the inhibition of key enzymes involved in both cytosolic and mitochondrial energy generation. In this study, the extent and pattern of metabolism of D-[U-14C, 5-3H]glucose was examined in perfused lungs isolated from rats before and after 24 h of in vivo exposure to 100% O2. Lung ATP levels after O2 exposure were maintained by a 53% increase in glucose utilization from an unexposed control value of 18.0 +/- 3.2 to 27.5 +/- 3.0 mumol 3H2O.h-1.g dry wt-1, accounted for by an enhanced rate of lactate plus pyruvate production from 15.7 +/- 2.0 to 32.7 +/- 4.1 mumol.h-1.g dry wt-1 with no alteration in lactate-to-pyruvate ratio. CO2 production was unaltered from a control rate of 27.5 +/- 4.0 14CO2 mumol.h-1.g dry wt-1. Maximal rates of glucose metabolism were determined by perfusion with 0.8 mM dinitrophenol, giving for air-exposed lungs a rate of 53.5 +/- 5.0 mumol 3H2O.h-1.g dry wt-1 and increased lactate plus pyruvate and 14CO2 production rates of 46.5 +/- 6.5 and 128.3 +/- 19.6 mumol.h-1.g dry wt-1, respectively. Although this maximal rate of glucose utilization was unaltered in oxygen-exposed lungs, lactate plus pyruvate production was further increased to 80.0 +/- 9.1 mumol.h-1.g dry wt-1 with a concomitant decrease in the dinitrophenol-induced rate of 14CO2 production to 81.5 +/- 9.2 mumol.h-1.g dry wt-1.(ABSTRACT TRUNCATED AT 250 WORDS)