Butylated hydroxytoluene (BHT)-induced lung damage in mice is an excellent model system for studying mechanisms of chemically-induced, reversible alveolar injury. Changes in the pulmonary contents of protein kinase C (PKC) and the calcium-dependent protease, calpain, were previously noted during the repair phase following BHT-induced pneumotoxicity. Calpain is believed to initiate PKC down-regulation. PKC-alpha is the major PKC isozyme and calpain II the major calpain isozyme in mouse lung. We have now studied the time course of these enzymatic changes in detail. Pulmonary PKC-alpha concentrations decreased as early as 45 min after an i.p. injection of 200 mg/kg BHT. Calpain II levels rose within the first 40 min after BHT injection, and then declined below control levels. The rapidity of these changes implies a role of these enzymes in mediating the onset of injury. Lung damage and repair, as estimated by measuring the lung weight/body weight ratio, is maximal 6 days after administration of this dose of BHT. The extent of the decreased PKC-alpha and calpain II concentrations at this time was linearly related to the estimated degree of injury based on increased lung weight. This correlation suggests the value of monitoring these enzymes as putative early biomarkers of alveolar injury.