Resistance of rat pulmonary alveolar epithelial cells to neutrophil- and oxidant-induced injury.

Abstract

We have previously reported that rat pulmonary alveolar epithelial cells are resistant to neutrophil-generated oxidants in contrast to the situation described for endothelial cells. In the present study, we investigated the roles of intracellular catalase and glutathione-dependent reactions in providing protection against cytotoxic concentrations of H2O2 and stimulated neutrophils. Catalase was found to be instrumental in protecting epithelial cells because when inhibited by either azide or 3-amino-1,2,4-triazole, there was an increase in the cytotoxic effect of exogenous H2O2 and stimulated neutrophils. Associated with this potentiation of injury was a reduction in epithelial cell clearance of H2O2. Partial inhibition of glutathione-dependent reactions by depleting intracellular glutathione with buthionine sulfoximine or by inhibiting the enzyme glutathione reductase with 1,3-bis(2-chloroethyl)-1-nitrosourea also augmented the cytotoxic effect of both H2O2 and stimulated neutrophils. This increase in neutrophil-induced cytotoxicity was caused by the addition of an oxidant-dependent mechanism of killing on top of the previously described oxidant-independent pathway. Importantly, the increased susceptibility to injury caused by inhibition of glutathione-dependent reactions was not associated with a reduction in epithelial cell consumption of exogenous H2O2, contrary to the case with catalase. This suggests that there are glutathione-dependent reactions that protect epithelial cells in ways separate from reducing the total burden of exogenous H2O2 on the cells.