Endotoxin administration in rats produced a significant increase in plasma, hepatic, and intestinal phospholipase A2 activity within three minutes after injection. The elevated phospholipase A2 activity seen in these tissues returned to normal levels six minutes after injection. The changes in phospholipase A2 activity were dose-dependent within the 0, 10, and 20 mg/kg range of treatment with Escherichia coli endotoxin. This increase in plasma, hepatic, and intestinal phospholipase A2 was abolished by prior treatment of the rats with 3 mg/kg indomethacin, a drug known to improve survival in endotoxic shock. The fact that the change in phospholipase A2 occurs soon after endotoxin administration and that the change in phospholipase is blocked by protective doses of indomethacin suggests that phospholipase A2 activation may be an important initial event in the lethal action of endotoxin, and that the protective effects of indomethacin may be directly related to inhibition of phospholipase A2 activity. Further, in vitro studies of the effects of indomethacin on hepatic phospholipase A2 activity showed that indomethacin significantly inhibited this enzyme. Indomethacin (25 μmol/L) produced 56% inhibition in phospholipase A2 activity and the apparent Ki for indomethacin was 9.2 μmol/L. Kinetic analysis using the Lineweaver-Burk method showed that the indomethacin inhibition was of the noncompetitive type.