Acute hypoxia alters eicosanoid production of perfused pulmonary artery endothelial cells in culture

Abstract

Hypoxia alters vascular tone which regulates regional blood flow in the pulmonary circulation. Endothelial derived eicosanoids alter vascular tone and blood flow and have been implicated as modulators of hypoxic pulmonary vasoconstriction. Eicosanoid production was measured in cultured bovine pulmonary endothelial cells during constant flow and pressure perfusion at two oxygen tensions (hypoxia: 4% O 2, 5% CO 2, 91% N 2; normoxia: 21% O 2, 5% CO 2, 74% N 2). Endothelial cells were grown to confluence on microcarrier beads. Cell cartridges ( N=8) containing 2 ml of microcarrier beads (≅ 5 × 10 6 cells) were constantly perfused (3 ml/min) with Krebs' solutions (pH 7.4, T 37°C) equilibrated with each gas mixture. After a ten minute equilibration period, lipids were extracted (C 18 Sep Pak®) from twenty minute aliquots of perfusate over three hours (nine aliquots per cartridge). Eicosanoids (6-keto PGF 1 α ; TXB 2; and total leukotriene [LT - LTC 4, LTD 4, LTE 4, LTF 4]) were assayed by radioimmunoassay. Eicosanoid production did not vary over time. 6-keto PGF 1 α production was increased during hypoxia (normoxia 291 ± 27 vs hypoxia 395 ± 35 ng/min/gm protein; p < 0.01). Thromboxane production (normoxia 19 ± 2 vs hypoxia 20 ± 2 ng/min/gm protein) and total leukotriene production (normoxia 363 ± 35 vs hypoxia 329 ± 29 ng/min/gm protein) did not change with hypoxia. These data demonstrated that oxygen increased endothelial prostacyclin production but did not effect thromboxane or leukotriene production.