Download PDF
Thromb Haemost 1986; 55(03): 369-374
DOI: 10.1055/s-0038-1661566
Original Article
Schattauer GmbH Stuttgart

Modulation of Arachidonic Acid Metabolism in Human Endothelial Cells by Glucocorticoids[*]

Raffaele De Caterina
Cornell University Medical College, New York, N. Y., USA
,
Babette B Weksler
Cornell University Medical College, New York, N. Y., USA
› Author Affiliations
Further Information

Publication History

Received 12 December 1985

Accepted 10 April 1986

Publication Date:
18 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

To learn whether glucocorticoids inhibit prostaglandin (PG) production in vascular endothelial cells, we investigated the effects of glucocorticoids on PG synthesis by cultured human umbilical vein endothelial cells (EC). Pretreatment of EC with dexamethasone (DX, 10-9 to 5 x 10-5 M) caused a dose-dependent inhibition of PGI2 production when PG synthesis from endogenous arachidonate was stimulated by human thrombin (0.25-2 U/ml) or ionophore A 23187 (1-5 μM). The inhibition was detectable at 10-7 M DX and maximal at 10-5 M (4.0 ± 0.7 vs. control: 7.7 ± 1.9 ng/ml, mean ± S.D., P <0.01). The production of PGE2 and the release of radiolabelled arachidonate (AA) from prelabelled cells were similarly inhibited. Prolonged incubation of EC with glucocorticoids was required to inhibit PG production or arachidonate release: ranging from 8% inhibition at 5 h to 44% at 38 h. In contrast, prostaglandin formation from exogenous AA was not altered by DX treatment. When thrombin or ionophore-stimulated EC were restimulated with exogenous AA (25 μM), DX-treated cells released more PGI2 than control cells (5.7 ± 0.5 vs. 4.1 ± 0.6 ng/ml, P <0.01). Both the decrease in PGI2 production after thrombin/ionophore and the increase after re-stimulation with AA were blunted in the presence of the protein synthesis inhibitor cycloheximide (0.1-0.2 μg/ml). Thus, incubation of EC with glucocorticoids inhibits PG production at the step of phospholipase activation. The time requirement for these steroid effects and their blunting by cycloheximide are consistent with the induction of regulatory proteins, possibly lipocortins, in endothelial cells.

Keywords

Glucocorticoids - Endothelial cells - Prostaglandins - Prostacyclin - Phospholipases

* Presented in abstract form at the 10th International Congress on Thrombosis and Haemostasis, San Diego, July 15-19, 1985.


 

  • References

  • 1 Haynes Jr RC, Murad F. Adrenocorticotropic hormone; adrenocortical steroids and their synthetic analogs; inhibitors of adrenocortical steroid biosynthesis. In The Pharmacological Basis of Therapeutics Goodman-Gilman A, Goodman LS, Gilman A. (eds.) 6th ed. pp 1466-1496 MacMillan Publishing Co; New York: 1980
  • 2 Lewis GP, Piper PJ. Inhibition of release of prostaglandins as an explanation of some of the actions of anti-inflammatory corticosteroids. Nature 1975; 254: 308-311
    CrossrefPubMedGoogle Scholar
  • 3 Hong SL, Levine L. pInhibition of arachidonic acid release from cells as the biochemical action of anti-inflammatory corticosteroids. Proc Natl Acad Sci U.S.A. 1976; 73: 1730-1734
    CrossrefPubMedGoogle Scholar
  • 4 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 5 Levin RI, Jaffe EA, Weksler BB, Tack-Goldman K. Nitroglycerin stimulates synthesis of prostacyclin by human endothelial cells. J Clin Invest 1981; 67: 762-769
    CrossrefPubMedGoogle Scholar
  • 6 Snedecor GW, Cochran WG. Statistical Methods. 2nd edition Iowa State University Press; Ames: 1980: 232-233
    Google Scholar
  • 7 De Caterina R, Dorso CR, Tack-Goldman K, Weksler BB. Nitrates and endothelial prostacyclin production: studies in vitro. Circulation 1985; 71: 176-182
    CrossrefPubMedGoogle Scholar
  • 8 Greaves MW, McDonald-Gibson W. Inhibition of prostaglandin biosynthesis by corticosteroids. Br Med J 1972; 2: 83-84
    CrossrefPubMedGoogle Scholar
  • 9 Tashjian Jr AH, Voelkel EF, McDonough J, Levine L. Hydrocortisone inhibits prostaglandin production by mouse fibrosarcoma cells. Nature 1975; 258: 739-741
    CrossrefPubMedGoogle Scholar
  • 10 Kantrowitz F, Robinson DR, McGuire MB, Levine L. Corticosteroids inhibit prostaglandin production by rheumatoid synovia. Nature 1975; 258: 737-739
    CrossrefPubMedGoogle Scholar
  • 11 Hirata F, Corcoran BA, Venkatasubramanian K, Schiffmann E, Axelrod J. Chemoattractants stimulate degradation of methylated phospholipids and release of arachidonic acid in rabbit leukocytes. Proc Natl Acad Sci U.S.A. 1979; 76: 2640-2643
    CrossrefPubMedGoogle Scholar
  • 12 Chandrabose KA, Lapetina EG, Schmitges CJ, Siegel MI, Cuatrecasas P. Action of corticosteroids in regulation of prostaglandin biosynthesis in cultured fibroblasts. Proc Natl Acad Sci U.S.A. 1978; 75: 214-217
    CrossrefPubMedGoogle Scholar
  • 13 Mitchell MD, Carr BR, Mason JI, Simpson ER. Prostaglandin biosynthesis in the human fetal adrenal gland: regulation by glucocorticosteroids. Proc Natl Acad Sci U.S.A. 1982; 79: 7547-7551
    CrossrefPubMedGoogle Scholar
  • 14 Bray MA, Gordon D. Prostaglandin production by macrophages and the effect of anti-inflammatory drugs. Br J Pharmacol 1978; 63: 635-642
    CrossrefPubMedGoogle Scholar
  • 15 Blackwell GJ, Carnuccio R, Di Rosa M, Flower RJ, Parente L, Persico P. Macrocortin: a polypeptide causing the antiphospholipase effect of glucocorticoids. Nature 1980; 287: 147-149
    CrossrefPubMedGoogle Scholar
  • 16 Cloix JF, Colard O, Rothhut B, Russo-Marie F. Characterization and partial purification of renocortins: two polypeptides formed in renal cells causing the anti-phospholipase action of glucocorticoids. Br J Pharmacol 1983; 79: 313-321
    CrossrefPubMedGoogle Scholar
  • 17 Willis AL, Davidson P, Ramwell PW, Brocklehurst WE, Smith B. Release and actions of prostaglandins in inflammation and fever: inhibition by antiinflammatory and anti-pyretic drugs. In Prostaglandins in Cellular Biology Ramwell PW, Pharriss BB. (eds.) pp 227-68 Plenum Press; New York: 1972
  • 18 Gryglewski RJ, Panczenko B, Korbut R, Grodzinska L, Ocetkiewicz A. Corticosteroids inhibit prostaglandin release from perfused mesenteric blood vessels of rabbit and from perfused lungs of sensitized guinea pig. Prostaglandins 1975; 10: 343-345
    CrossrefPubMedGoogle Scholar
  • 19 Weksler BB, Marcus AJ, Jaffe EA. Synthesis of prostaglandin I2 (prostacyclin) by cultured human and bovine endothelial cells. Proc Natl Acad Sci U.S.A. 1977; 74: 3922-3926
    CrossrefPubMedGoogle Scholar
  • 20 Ager A, Pearson JD, Gordon JL. Radioimmunoassay of 6-oxo-prostaglandin F1α and prostaglandin E2 produced by pig aortic endothelium in culture. Biochem Soc Trans 1978; 7: 1065-1066
    PubMedGoogle Scholar
  • 21 Buchanan MR, Butt RW, Magas Z, Van Ryn J, Hirsh J, Nazir DJ. Endothelial cells produce a lipooxygenase derived chemo-repellent which influences platelet/ endothelial cell interactions. Effects of aspirin and salicylate. Thromb Haemostas 1985; 53: 306-311
    PubMedGoogle Scholar
  • 22 Weksler BB. Prostaglandins and vascular function. Circulation 1984; 70 III 63-71
    CrossrefPubMedGoogle Scholar
  • 23 MacIntyre DE, Pearson JD, Gordon JL. Localization and stimulation of prostacyclin production in vascular cells. Nature 1978; 271: 549-551
    CrossrefPubMedGoogle Scholar
  • 24 Jose PJ, Page DA, Wolstenholme BE, Williams TJ, Dumonde DC. Bradykinin-stimulated prostaglandin E2 production by endothelial cells and its modulation by anti-inflammatory compounds. Inflammation 1981; 5: 363-378
    CrossrefPubMedGoogle Scholar
  • 25 McCall E, Ingram A, Bagley Jr GC. Anti-inflammatory steroids inhibit prostaglandin production by cultured endothelial cells. Fed Proc 1982; 41: 455 (Abstr)
    PubMedGoogle Scholar
  • 26 Huck RL, Whorton AR, Young SL. Corticosteroids (CS) do not inhibit arachidonic acid (AA) and prostacyclin (PGI2) release from cultured porcine aortic endothelial cells (EC). Fed Proc 1982; 41: 1642 (Abstr)
    PubMedGoogle Scholar
  • 27 Blajchman MA, Senyi AF, Hirsh J, Surya Y, Buchanan M, Mustard JF. Shortening of the bleeding time in rabbits by hydrocortisone caused by inhibition of prostacyclin generation by the vessel wall. J Clin Invest 1979; 63: 1026-1035
    CrossrefPubMedGoogle Scholar
  • 28 Jorensen KA, Stoffersen E. Hydrocortisone inhibits platelet prostaglandin and endothelial prostacyclin production. Pharmacol Res Comm 1981; 13: 579-586
    CrossrefPubMedGoogle Scholar
  • 29 Hirata F, Schiffmann E, Venkatasubramanian K, Salomon D, Axelrod J. A phospholipase A2 inhibitory protein in rabbit neutrophils induced by glucocorticoids. Proc Natl Acad Sci U.S.A. 1980; 77: 2533-2536
    CrossrefPubMedGoogle Scholar
  • 30 Russo-Marie F, Duval D. Dexamethasone-induced inhibition of prostaglandin production does not result from a direct action on phospholipase activities but is mediated through a steroid-inducible factor. Biochim Biophys Acta 1982; 712: 177-185
    CrossrefPubMedGoogle Scholar
  • 31 Piovella F, Giddings JC, Almasio P, Ricetti MM, Thomas JE. Effects of ticlopidine and dexamethasone on fibronectin and Factor VIII-related antigen synthesis by cultured endothelial cells. Thromb Res 1983; Suppl IV: 69-73
    PubMedGoogle Scholar
  • 32 Thomas JM F, Hullin F, Chap H, Douste-Blazy L. Phosphatidylcholine is the major phospholipid providing arachidonic acid for prostacyclin synthesis in thrombin-stimulated human endothelial cells. Thromb Res 1984; 34: 117-123
    CrossrefPubMedGoogle Scholar
  • 33 Thomas JM F, Chap H, Douste-Blazy L. Calcium ionophore A23187 induces arachidonic acid release from phosphatidylcholine in cultured human endothelial cells. Biochem Biophys Res Comm 1981; 103: 819-824
    CrossrefPubMedGoogle Scholar
  • 34 Danon A, Assouline G. Inhibition of prostaglandin biosynthesis by corticosteroids requires RNA and protein synthesis. Nature 1978; 273: 552-554
    CrossrefPubMedGoogle Scholar
  • 35 Di Rosa M, Flower RJ, Hirata F, Parente L, Russo-Marie F. Nomenclature announcement. Anti-phospholipase proteins. Prostaglandins 1984; 28: 441-442
    CrossrefPubMedGoogle Scholar
  • 36 Egan RL, Paxton J, Kuehl FA. Mechanism for irreversible self-deactivation of prostaglandin synthetase. J Biol Chem 1976; 251: 7329-7335
    PubMedGoogle Scholar
  • 37 Egan RW, Gale PH, Baptista EM, Kennicott KL, VandenHeuvel WJ A, Walker RW, Fagerness PE, Kuehl Jr FA. Oxidation reactions by prostaglandin cyclooxygenase-hydroperoxidase. J Biol Chem 1981; 256: 7352-7361
    PubMedGoogle Scholar
  • 38 Ham EA, Egan RW, Soderman DD, Gale PH, Kuehl Jr FA. Peroxidase-dependent deactivation of prostacyclin synthetase. J Biol Chem 1979; 254: 2191-2194
    PubMedGoogle Scholar
  • 39 Brotherton AF A, Hoak JC. Prostacyclin biosynthesis in cultured vascular endothelium is limited by deactivation of cyclooxygenase. J Clin Invest 1983; 72: 1255-1261
    CrossrefPubMedGoogle Scholar
  • 40 Naray-Fejes-Toth A, Fejes-Toth G, Fischer C, Froelich JC. Effects of dexamethasone on in vivo prostanoid production in the rabbit. J Clin Invest 1984; 74: 120-123
    CrossrefPubMedGoogle Scholar
  • 41 Casey ML, MacDonald PC, Mitchell MD. Despite a massive increase in cortisol secretion in women during parturition, there is an equally massive increase in prostaglandin synthesis. A paradox?. J Clin Invest 1985; 75: 1852-1857
    CrossrefPubMedGoogle Scholar
  • 42 Schleimer RP, Schulman ES, MacGlashan Jr DW, Peters SP, Hayes EC, Adams GK III, Lichtenstein LM, Adkinson Jr AF. Effects of dexamethason on mediator release from human lung fragments and purified human lung mast cells. J Clin Invest 1983; 71: 1830-1835
    CrossrefPubMedGoogle Scholar
  • 43 Marcus AJ, Weksler BB, Jaffe EA, Broekman MI. Synthesis of prostacyclin from platelet-derived endoperoxides by cultured human endothelial cells. J Clin Invest 1980; 66: 979-986
    CrossrefPubMedGoogle Scholar
  • 44 FitzGerald GA, Brash AR, Oates JA, Pedersen AK. Endogenous prostacyclin biosynthesis and platelet function during selective inhibition of thromboxane synthase in man. J Clin Invest 1983; 72: 1336-1343
    CrossrefPubMedGoogle Scholar
  • 45 Crutchley DJ, Ryan US, Ryan JW. Glucocorticoid modulation of prostacyclin production in cultured bovine pulmonary endothelial cells. J Pharmacol Exper Ther 1985; 233: 650-655
    PubMedGoogle Scholar
  • 46 Mendelsohn FA O, Lloyd CJ, Kachel C, Funder JW. Induction by glucocorticoids of angiotensin-converting enzyme production from bovine endothelial cells in culture and rat lung in vivo. J Clin Invest 1982; 70: 684-692
    CrossrefPubMedGoogle Scholar