Summary
The effect of the stable prostacyclin (PGI2) mimetic iloprost on cyclic AMP levels was investigated in cultured porcine aortic endothelial cells. Iloprost (10−10−10−5 mol/l) did not change cyclic AMP levels at passage 1 when endothelial cells were untreated but did so after inhibition of endogenous PGI2 formation by 48 or 72 h treatment with indomethacin or diclofenac (10−5 mol/l). Iloprost increased cyclic AMP in a concentration-dependent manner and up to 6-fold above control when cells from passage 6 were used. In these cells, basal PGI2 generation was reduced to 20% of that at passage 1. Cyclic AMP stimulation by iloprost (10−5 mol/l) in passage 6 cells was enhanced, reaching up to 11-fold the control level, when cells were cultured for 48 h in the presence of indomethacin or diclofenac (10−5 mol/l). Cyclic AMP formation in LLC-PK1 cells, a kidney epithelial cell line without endogenous PGI2 biosynthesis, was markedly (25-fold above basal) stimulated by iloprost and unchanged by pretreatment with indomethacin and diclofenac. The data demonstrate that a continuous basal PGI2 generation occurs in porcine aortic endothelial cells that may be sufficient to completely desensitize PGI2-dependent adenylate cyclase activation, presumably at the receptor or GTP-binding protein level.
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References
Alt U, Leigh PW, Wilkins AJ, Morris PK, MacDermot J (1986) Desensitization of iloprost responsiveness in human platelets follows prolonged exposure to iloprost in vitro. Br J Clin Pharmacol 22:118–119
Ashby B, Almonor GO, Wernick E, Selak MA (1991) Prostaglandin-concentration-dependent desensitization of adenylate cyclase in human erythroleukaemia (HEL) cells is abolished by pertussis toxin and enhanced by dimethyl sulphoxide. Biochem J 280:801–804
Brotherton AFA, Hoak JC (1982) Role of Ca2+ and cyclic AMP in the regulation of the production of PGI2 by the vascular endothelium. Proc Natl Acad Sci USA 79:495–499
Crutchley DJ, Hirsh MJ (1991) The stable prostacyclin analog, iloprost, and prostaglandin El inhibit monocyte procoagulant activity in vitro. Blood 78:382–386
Friedl A, Harmening C, Schuricht B, Hamprecht B (1985) Rat atrial natriuretic peptide elevates the level of cyclic GMP in astroglia-rich brain cell cultures. Eur J Pharmacol 111:141–142
Gryglewski RJ, Moncada S, Palmer RMJ (1986) Bioassay of prostacyclin and endothelium-derived relaxing factor (EDRF) from porcine aortic endothelial cells. Br J Pharmacol 87:685–694
Hopkins NK, Gorman RR (1981) Regulation of endothelial cell cyclic nucleotide metabolism by prostacyclin. J Clin Invest 67:540–546
Jaschonek K, Faul C, Schmidt H, Renn W (1988) Desensitization of platelets to iloprost. Loss of specific binding sites and heterologous desensitization of adenylate cyclase. Eur J Pharmacol 147:187–196
Keen M, Kelly E, MacDermot J (1989) Prostaglandin receptors in the cardiovascular system: Potential selectivity from receptor subtypes or modified responsiveness. Eicosanoids 2:193–197
Kuhn M; Otten A, Frölich JC, Förstermann U (1991) Endothelial cyclic GMP and cyclic AMP do not regulate the release of endothelium-derived relaxing factor/nitric oxide from bovine aortic endothelial cells. J Pharmacol Exp Ther 256:677–682
Lückhoff A, Mülsch A, Busse R (1990) cAMP attenuates autacoid release from endothelial cells: Relation to internal calcium. Am J Physiol 258:H960-H966
Martin W, White DG, Henderson AH (1988) Endothelium-derived relaxing factor and atriopeptin II elevate cyclic GMP levels in pig aortic endothelial cells. Br J Pharmacol 93:229–239
Pearson JD, Carleton JS, Huchings A (1983) Prostacyclin release stimulated by thrombin or bradykinin in porcine endothelial cells cultured from aorta and umbilical vein. Thromb Res 29:115–125
Schafer AI, Gimbrone MA Jr, Handin RI (1980) Endothelial cell adenylate cyclase: Activation by catecholamines and prostaglandin I2. Biochem Biophys Res Commun 96:1640–1647
Schröder H, Schrör K (1989) Cyclic GMP stimulation by vasopressin in LLC-PK1I cells is L-arginine dependent. Naunyn-Schmiedeberg's Arch Pharmacol 340:475–477
Schröder H, Strobach H, Schrör K (1992) Nitric oxide but not prostacyclin is an autocrine endothelial mediator. Biochem Pharmacol 43:533–537
Schrör K, Seidel H (1988) Blood-vessel wall arachidonate metabolism and its pharmacological modification in a new in vitro assay system. Naunyn-Schmiedeberg's Arch Pharmacol 337:177–182
Schrör K, Sauerland S, Kuhn A, Rosen R (1980) Different sensitivities of prostaglandin cyclooxygenases in blood platelets and coronary arteries against non-steroidal antiinflammatory drugs. Naunyn-Schmiedeberg's Arch Pharmacol 313:69–76
Steiner AL, Parker CW, Kipnis DM (1972) Radioimmunoassay for cyclic nucleotides. J Biol Chem 247:1106–1113
Voyta JC, Via DP, Butterfield CE, Zetter BR (1984) Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J Cell Biol 99:2034–2040
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Schröder, H., Schrör, K. Prostacyclin-dependent cyclic AMP formation in endothelial cells. Naunyn-Schmiedeberg's Arch Pharmacol 347, 101–104 (1993). https://doi.org/10.1007/BF00168779
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DOI: https://doi.org/10.1007/BF00168779