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S35b, a new phenylsulfonylfuroxan compound, inhibits thrombin-induced synthesis of platelet-activating factor and prostacyclin in human endothelial cells

  • Inflammation
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Abstract

Endothelial cells (EC) produce platelet activating factor (PAF) and prostacyclin (PGI2) in response to inflammatory agents such as thrombin. Upon cell stimulation a calcium-dependent phospholipase A2 (PLA2) is activated which hydrolyzes a membrane phospholipid to yield 1-0-alkyl-2-lyso-sn-glycero-3-phospho-choline (lyso-PAF) and free arachidonic acid. Lyso-PAF is in turn converted into PAF by a specific acetyltransferase and arachidonic acid is metabolized via cyclic endoperoxides to PGI2. In the present study we report that S35b (4-methyl-3-phenylsulfonylfuroxan), an new phenyl-sulfonylfuroxan compound with potent antiaggregatory effect, inhibits thrombin-induced PAF synthesis and acetyltransferase activation as well as PGI2 production in human umbilical vein endothelial cells (HUVEC) in a concentration dependent way. Additionally, we show that S35b stimulates the production of cyclic GMP (cGMP) in HUVEC in a concentration- and time-dependent manner. At high concentration, S35b potentiates the cAMP increase induced by iloprost or forskolin without having a significant influence on cAMP level itself. Potentiation of cAMP increase during agonist-induced EC stimulation seems not to be important for the effect of S35b on cellular function as the compound is active in inhibiting PAF production when endothelial cells are pretreated with indomethacin to block PGI2 synthesis. The increase of cGMP evoked by S35b may account for the effect on endothelial cell function.

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References

  1. G. Camussi, M. Aglietta, F. Malavasi, C. Tetta, W. Piacibello, F. Sanavio and F. Bussolino,The release of platelet-activating factor from human endothelial cells in culture. J. Immunol.131, 2397–2403 (1983).

    PubMed  Google Scholar 

  2. S. M. Prescott, G. A. Zimmerman and T. M. McIntyre,Human endothelial cells in culture produce platelet-activating factor (1-alkyl-2-acetyl-sn-glyceryl-3-phosphocholine) when stimulated with thrombin. Proc. Natl. Acad. Sci. USA81, 3934–3938 (1984).

    PubMed  Google Scholar 

  3. T. M. McIntyre, G. A. Zimmerman, K. Satoh and S. M. Prescott,Cultured endothelial cells synthesize both platelet-activating factor and prostacyclin in response to histamine, bradykimin and adenosine triphosphate. J. Clin. Invest.76, 271–280 (1985).

    PubMed  Google Scholar 

  4. E. A. Jaffe, J. Grulich, B. B. Weksler, G. Hampel and K. Watanabe,Correlation between thrombin-induced prostacyclin production and inositol trisphosphate and cytosolic free calcium levels in cultured human endothelial cells. J. Biol. Chem.262, 9557–8565 (1987).

    Google Scholar 

  5. T. J. Hallam, J. D. Pearson and L. A. Needham,Thrombin-stimulated elevation of human endothelial-cell cytoplasmic free calcium concentration causes prostacyclin production. Biochem. J.251, 243–249 (1988).

    PubMed  Google Scholar 

  6. P. Braquet, T. Y. Shen, L. Touqui and B. B. Vargaftig,Perspectives in platelet-activating factor research. Pharmacol. Rev.39, 97–145 (1987).

    PubMed  Google Scholar 

  7. S. M. Prescott, G. A. Zimmerman and T. M. McIntyre,Plateletactivating factor. J. Biol. Chem.265, 17381–17384 (1990).

    PubMed  Google Scholar 

  8. F. Snyder,Platelet-activating factor and related acetylated lipids as potent biologically active cellular mediators. Am. J. Physiol.259, C697-C708 (1990).

    PubMed  Google Scholar 

  9. G. A. Zimmerman, T. M. McIntyre, M. Mehra and S. M. Prescott,Endothelial cell-associated platelet-activating factor: a novel mechanism for signaling intercellular adhesion. J. Cell Biol.110, 529–540 (1990).

    Article  PubMed  Google Scholar 

  10. R. Gryglewski, S. Bunting, S. Moncada, R. Flower and J. R. Vane,Mediators produced by the endothelial cell. Hypertension12, 530–548 (1988).

    PubMed  Google Scholar 

  11. J. R. Vane,Regulatory function of the vascular endothelium. N. Engl. J. Med.323, 27–36 (1990).

    PubMed  Google Scholar 

  12. S. M. Prescott, T. M. McIntyre and G. A. Zimmerman,The role of platelet-activating factor in endothelial cells. Thromb. Haemostas.64, 99–103 (1990).

    Google Scholar 

  13. M. Hirafuji, J. M. Mencia-huerta and J. Benveniste,Regulation of PAF-acether (platelet-activating factor) biosynthesis in cultured human vascular endothelial cells stimulated with thrombin. Biochim. Biophys. Acta,930, 359–369 (1987).

    Article  PubMed  Google Scholar 

  14. D. Ghigo, F. Bussolino, G. Garbarino, R. Heller, F. Turrini, G. P. Pescarmona, E. J. Cragoe Jr., L. Pegoraro and A. Bosia,Role of Na +/H + exchange in thrombin-induced platelet-activating factor production by human endothelial cells. J. Biol. Chem.263, 19437–19446 (1988).

    PubMed  Google Scholar 

  15. R. E. Whatley, P. Nelson, G. A. Zimmerman, D. L.Stevens, C. J. Parker, T. M. McIntyre and S. M. Prescott,The regulation of platelet-activating factor production in endothelial cells: the role of calcium and protein kinase. C. J. Biol. Chem.264, 6325–6333 (1989).

    Google Scholar 

  16. G. B. Zavoico, J. K. Hrbolich, M. A. Gimbrone and A. I. Schafer,Enhancement of thrombin- and ionomycin-stimulated prostacyclin and platelet-activating factor production in cultured endothelial cells by a tumor-promoting phorbol ester. J. Cell. Physiol.143, 596–605 (1990).

    Article  PubMed  Google Scholar 

  17. R. Heller, F. Bussolino, D. Ghigo, G. Garbarino, H. Schröder, G. Pescarmona and A. Bosia,Protein kinase C and cyclic AMP modulate thrombin-induced platelet-activating factor synthesis in human endothelial cells. Biochim. Biophys. Acta1093 55–64 (1991).

    Article  PubMed  Google Scholar 

  18. F. H. Chilton and T. R. Connell,1-Ether-linked phosphoglycerides, Major endogenous sources of arachidonate in the human neutrophil. J. Biol. Chem.263, 5260–5265 (1988).

    PubMed  Google Scholar 

  19. R. E. Whatley, G. A. Zimmerman, T. M. McIntyre, R. Taylor and S. M. Prescott:Production of platelet-activating factor by endothelial cells. Semin. Thromb. Hemostas. 13, 445–453 (1987).

    Google Scholar 

  20. B. B. Weksler, A. J. Markus and E. A. Jaffe,Synthesis of prostaglandin I 2 (prostacyclin) by cultured human and bovine endothelial cells. Proc. Natl. Acad. Sci. USA74, 3922–3926 (1977).

    PubMed  Google Scholar 

  21. A. I. Schafer, D. D. Crawford and M. A. Gimbrone,Unidirectional transfer of prostaglandin endoperoxides between platelets and endothelial cells. J. Clin. Invest.73, 1105–1108 (1984).

    PubMed  Google Scholar 

  22. M. G. Lampugnani, M. Pedenovi, E. Dejana, D. Rotilio, M. B. Donati, F. Bussolino, G. Garbarino, D. Ghigo and A. Bosia,Human α-thrombin induces phosphoinositide turnover and Ca ++ movements in cultured human umbilical vein endothelial cells. Thromb. Res.54, 75–87 (1989).

    Article  PubMed  Google Scholar 

  23. M. J. Berridge,Inositol trisphosphate and diacylglycerol, two interacting second messengers. Annu. Rev. Biochem.56, 159–193 (1987).

    PubMed  Google Scholar 

  24. R. Heller, F., Bussolino, D. Ghigo, G. Garbarino, G. P. Pescarmona, U. Till and A. Bosia,Nitrovasodilators inhibit thrombin-induced platelet-activating factor synthesis in human endothelial cells. Biochem. Pharmacol.44, 223–229 (1992).

    Article  PubMed  Google Scholar 

  25. D. Ghigo, R. Heller, R. Calvino, P. Alessio, R. Fruttero, A. Gasco, A. Bosia and G. Pescarmona,Characterization of a new compound, S35b, as a guanylate cyclase activator in human platelets. Biochem. Pharmacol.43, 1281–1288 (1992).

    Article  PubMed  Google Scholar 

  26. A. L. Steiner, C. W. Parker and D. M. Kipnis,Radioimmunoassay for cyclic nucleotides. I. Preparation of antibodies and iodinated cyclic nucleotides. J. Biol. Chem.247, 1106–1113 (1972).

    PubMed  Google Scholar 

  27. E. G. Bligh and W. J. Dyer,A rapid method of total lipid extraction and purificaton. Can. J. Biochem. Physiol.37, 911–917 (1959).

    PubMed  Google Scholar 

  28. G. A. Zimmerman, R. E. Whatley, T. M. McIntyre, D. M. Benson and S. M. Prescott,Endothelial cells for studies of plateletactivating factor and arachidonate metabolites. In Methods in Enzymology, Vol.187, (Ed R. C. Murphy and F. A. Fitzpatrick) pp. 520–535. Academic Press Inc. 1990.

  29. F. Bussolino, F. Gremo, C. Tetta, G. P. Pescarmona and G. Camussi,Production of platelet-activating factor by chick retina. J. Biol. Chem.261, 16502–16508 (1986).

    PubMed  Google Scholar 

  30. F. Snyder, M. Blank, T.-C. Lee, M. Robinson and D. Woodard,Measurements of key enzyme activities involved in the metabolism of platelet activating factor. In Methods in Enzymology, Vol.141, (Ed. R. C. Murphy and F. A. Fitzpatrick) pp. 379–396, Academic Press Inc. 1987.

  31. A. M. Marcus, B. B. Weksler, E. A. Jaffe and M. J. Broekman,Synthesis of prostacyclin from platelet-derived endoperoxides by cultured human endothelial cells. J. Clin. Invest.66, 979–986 (1980).

    PubMed  Google Scholar 

  32. R. Calvino, R. Fruttero, D. Ghigo, A. Bosia, G. P. Pescarmona and A. Gasco,4-Methyl-3-(R-sulfonyl)-furoxans: a new class of potent inhibitors of the platelet aggregation. J. Med. Chem.35, 3296–3300 (1992).

    Article  PubMed  Google Scholar 

  33. C. Lugnier, P. Schoeffter, A. Le Bec, E. Strouthou and J. C. Stoclet,Selective inhibition of cyclic nucleotide phosphodiesterases of human, bovine and rat aorta. Biochem. Pharmacol.35, 1743–1751 (1986).

    Article  PubMed  Google Scholar 

  34. E. Noack and M. Feelisch,Molecular aspects underlying the vasodilator action of molsidomine. J. Cardiovasc. Pharmacol.14 (Suppl. 11) S1-S5 (1989).

    Google Scholar 

  35. M. Feelisch and E. Noack,Nitric oxide (NO) formation from nitrovasodilators occurs independently of hemoglobin or nonheme iron. Eur. J. Pharmacol.142, 465–469 (1987).

    Article  PubMed  Google Scholar 

  36. M. Feelisch, J. Ostrowski and E. Noack,On the mechanism of NO release from sydnonimines. J. Cardiovasc. Pharmacol.14 (Suppl. 11, S13-S22 (1989).

    Google Scholar 

  37. S. A. Waldman and F. Murad,Cyclic GMP synthesis and function. Pharmacol. Rev.39, 163–196 (1987).

    PubMed  Google Scholar 

  38. S. Nakashima, T. Tohmatsu, H. Hattori, Y. Okano and Y. Nozawa,Inhibitory action of cyclic GMP on secretion, polyphosphoinositide hydrolysis and calcium mobilization in thrombinstimulated human platelets. Biochem. Biophys. Res. Commun.135, 1099–1104 (1986).

    Article  PubMed  Google Scholar 

  39. M.-F. Simon, H. Chap and L. Douste-Blazy,Effect of a stimulant of guanylate cyclase, SIN-1, on calcium movements and phospholipase C activation in thrombin-stimulated human platelets. Biochem. Pharmacol.37, 1263–1269 (1988).

    Article  PubMed  Google Scholar 

  40. M. Hirata, K. P. Kohse, C.-H. Chang, T. Ikebe and F. Murad,Mechanism of cyclic GMP inhibition of inositol phosphate formation in rat aorta segments and cultured bovine aortic smooth muscle cells. J. Biol. Chem.265, 1268–1273 (1990).

    PubMed  Google Scholar 

  41. R. Deana, M. Ruzzene, M. G. Doni, F. Zoccarato and A. Alexandre,Cyclic GMP and nitroprusside inhibit the activation of human platelets by fluoroaluminate. Biochim. Biophys. Acta1014, 203–206 (1989).

    Article  PubMed  Google Scholar 

  42. R. O. Morgan and A. C. Newby,Nitroprusside differentially inhibits ADP-stimulated calcium influx and mobilization in human platelets. Biochem. J.258, 447–454 (1989).

    PubMed  Google Scholar 

  43. M.-F. Simon and H. Chap,Inhibition of calcium influx in thrombin-stimulated platelets by SIN-1, an activator of soluble guanylate cyclase. J. Cardiovasc. Pharmacol.14 (Suppl. 11), S106-S110 (1989).

    PubMed  Google Scholar 

  44. R. E. Whatley, D. F. Fennell, J. A. Kurrus, G. A. Zimmerman, T. M. McIntyre and S. M. Prescott,Synthesis of plateletactivating factor by endothelial cells: the role of G proteins. J. Biol. Chem.265, 15550–15559 (1990).

    PubMed  Google Scholar 

  45. D. H. Maurice and R. J. Haslam,Molecular basis of the synergistic inhibition of platelet function by nitrovasodilators and activators of adenylate cyclase: Inhibition of cyclic AMP breakdown by cyclic cGMP. Mol. Pharmacol.37, 671–681 (1990).

    PubMed  Google Scholar 

  46. K. Schrör, H. Darius, R. Matzky and R. Ohlendorf,The antiplatelet and cardiovascular actions of a new carbacyclin derivative (ZK 36374) — equipotent to PGI 2 in vitro. Naunyn-Schmiedeberg's Arch Pharmacol.316, 252–255 (1981).

    Article  Google Scholar 

  47. A. B. Legrand, T. K. Narayanan, U. S. Ryan, R. S. Aronstam and J. D. Catravas,Modulation of adenylate cyclase activity in cultured bovine pulmonary arterial endothelial cells. Biochem. Pharmacol.38, 423–430 (1989).

    Article  PubMed  Google Scholar 

  48. G. Doni, R. Deana, S.D. Bertoncello, F. Zoccarato and A. Alexandre,Forskolin and prostacyclin inhibit fouoride induced platelet activation and protein kinase C dependent reponses. Biochem. Biophys. Res. Commun.156, 1316–1323 (1988).

    Article  PubMed  Google Scholar 

  49. E. R. Lazarowski and E. G. Lapetina,Activation of platelet phospholipase C is inhibited by elevation of cyclic AMP. Biochem. Biophys. Res. Commun.158, 440–444 (1989).

    Article  PubMed  Google Scholar 

  50. H. Schröder, H. Strohbach and K. Schrör,Nitric oxide but not prostacyclin is an autocrine endothelial mediator. Biochem. Pharmacol.43, 533–537 (1992).

    Article  PubMed  Google Scholar 

  51. H. Schröder and K. Schrör,Prostacyclin-dependent cyclic AMP formation in endothelial cells. NS Arch. Pharmacol.347, 101–104 (1973).

    Google Scholar 

  52. Y. Yada, S. Nagao, S. Okano and Y. Nozawa,Inhibition by cyclic AMP of guanine nucleotide-induced activation of phosphoinositide-specific phospholipase C in human platelets. FEBS Lett.242, 368–372 (1989).

    Article  PubMed  Google Scholar 

  53. J. G. de Mey, M. Claeys and P. M. Vanhoutte,Endothelium-dependent inhibitory effects of acetylcholine, adenosine diphosphate, thrombin and arachidonic acid in the canine femoral artery. J. Pharm. Exp. Ther.222, 166–173 (1982).

    Google Scholar 

  54. R. M. J. Palmer, A. G. Ferridge and S. Moncada,Nitric oxide release accounts for the biological activity of endotheliumderived relaxing factor. Nature327, 524–526 (1987).

    PubMed  Google Scholar 

  55. W. Martin, D. G. White and A. H. Henderson,Endothelium-derived relaxing factor and atriopeptin II elevate cyclic GMP levels in pig aortic endothelial cells. Br. J. Pharmacol.93 229–239 (1988).

    PubMed  Google Scholar 

  56. C. Boulanger, V. B. Schini, S. Moncada and P. M. Vanhoutte,Stimulation of cyclic GMP production in cultured endothelial cells of the pig by bradykinin, adenosine diphosphate, calcium ionophore A23187 and nitric oxide. Br. J. Pharmacol.101, 152–156 (1990).

    PubMed  Google Scholar 

  57. G. M. Vercellotti, H. Q. Yin, K. S. Gustafson, R. D. Nelson and H. S. Jacob,Platelet-activating factor primes neutrophil responses to agonists: Role in promoting neutrophil-mediated endothelial damage. Blood71, 1100–1107 (1988).

    PubMed  Google Scholar 

  58. P. Kubes, G. Ibbotson, J. Russell, J. L. Wallace and D. N. Granger,Role of platelet-activating factor in ischemia/reperfusion-induced leukocyte adherence. Am. J. Physiol.259, G300-G305 (1990).

    PubMed  Google Scholar 

  59. F. A. Nicolini, P. Mehta, D. Lawson and J. L. Mehta,Reduction in human neutrophil chemotaxis by the prostacyclin analogue iloprost. Thromb. Res.59, 669–674 (1990).

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Pathological Biochemistry, Medical Academy, Erfurt, Germany

    R. Heller & U. Till

  2. Dipartimento di Genetica, Biologia e Chimica Medica, Dipartimento di Scienza e Tecnologia del Farmaco, University of Torino, Via Santena 5 bis, 10126, Torino, Italy

    R. Calvino, R. Fruttero & A. Gasco

  3. Dipartimento di Genetica, Biologia e Chimica Medica, University of Torino, Italy

    F. Bussolino, D. Ghigo, P. Alessio, R. Todde, G. Pescarmona & A. Bosia

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  1. R. Heller
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Heller, R., Bussolino, F., Calvino, R. et al. S35b, a new phenylsulfonylfuroxan compound, inhibits thrombin-induced synthesis of platelet-activating factor and prostacyclin in human endothelial cells. Agents and Actions 40, 157–165 (1993). https://doi.org/10.1007/BF01984055

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