Role of Ca2+ and cyclic AMP in the regulation of the production of prostacyclin by the vascular endothelium.
Incubation of primary monolayer cultures of human umbilical vein endothelial cells with buffer, thrombin (0.5 unit/ml), ionophore A23187 (10 microM), arachidonic acid (20 microM), prostaglandin H2 (PGH2) (4 microM) resulted in prostacyclin (PGI2) production in nanomolar quantities to the extent of 36 +/- 2, 276 +/- 13, 485 +/- 32, 533 +/- 22, and 532 +/- 22, respectively, as measured by radioimmunoassay of 6-keto-PGF alpha. Preincubation of the endothelium with 1 mM 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate, an antagonist of cytoplasmic Ca2+, or with 4 mM 1-methyl-3-isobutylxanthine (MIX), an inhibitor of cyclic nucleotide phosphodiesterase activity, blocked PGI2 release induced by thrombin or A23187, decreased arachidonic acid-induced release by approximately 50%, but had no effect on PGH2-induced release. Radioimmunoassay of cAMP in the endothelium showed that the basal level (1.85 +/- 0.14 pmol of cAMP per 4.5 x 10(5) cells) was increased by an average of 3.9-fold with 4 mM MIX. PGI2 (0.4 microM) had no significant effect on cAMP levels in the absence of MIX, but caused a 2-fold increase with 4 mM MIX. The findings suggest that: (i) the stimulation of PGI2 biosynthesis is mediated by Ca2+, (ii) increased cAMP inhibits PGI2 production, and (iii) cAMP phosphodiesterase activity modulates PGI2-induced increases in the intracellular concentration of cAMP.
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000502:physiology*; D004727:Endothelium / Q000378:metabolism*; D011464:Epoprostenol / Q000096:biosynthesis*; D006801:Humans; D011453:Prostaglandins / Q000096:biosynthesis*; D011463:Prostaglandins H / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology; D014471:Umbilical Veins
D001095:Arachidonic Acids; D011453:Prostaglandins; D011463:Prostaglandins H; D016718:Arachidonic Acid; D000001:Calcimycin; D011464:Epoprostenol; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D013917:Thrombin; D002118:Calcium