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. 1989 Nov;98(3):973–981. doi: 10.1111/j.1476-5381.1989.tb14628.x

Effects of amiloride analogues on the production of prostacyclin by aortic endothelial cells.

J M Boeynaems 1, D Demolle 1, C Lagneau 1, E J Cragoe Jr 1
PMCID: PMC1854786  PMID: 2511997

Abstract

1. The release of prostacyclin (PGI2) from bovine aortic endothelial cells stimulated by adenosine 5'-triphosphate (ATP) was decreased by amiloride analogues bearing alkyl groups on the 5-amino nitrogen atom, like 5-(N-ethyl-N-isopropyl)amiloride (EIPA), which are inhibitors of the Na+/H+ exchanger. Analogues substituted on a terminal guanidino nitrogen atom were not inhibitory. 2. The release of PGI2 induced by ATP was not significantly depressed in a Na+-poor medium or in a medium acidified to pH 6.9, two conditions known to inhibit the Na+/H+ exchanger. 3. Cytoplasmic alkalinization by ammonium chloride did not suppress the inhibitory action of EIPA. By itself, ammonium chloride decreased the response of endothelial cells to ionophore A23187 and ATP, whereas sodium acetate had no effect. 4. EIPA did not decrease the mobilization of free arachidonic acid induced by ATP. It inhibited the conversion of exogenous arachidonate into PGI2 and prostaglandin E2 (PGE2). 5. Although the intracellular pH was not measured in this study, it seems unlikely that cytoplasmic alkalinization via the activation of the Na+/H+ exchanger plays a significant role in the stimulatory action of ATP on the release of PGI2 from endothelial cells. The inhibition of that release by EIPA and other amiloride analogues might involve a direct effect on cyclo-oxygenase, although an action on the reacylation of free arachidonic acid cannot be excluded.

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Selected References

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  1. Besterman J. M., May W. S., Jr, LeVine H., 3rd, Cragoe E. J., Jr, Cuatrecasas P. Amiloride inhibits phorbol ester-stimulated Na+/H+ exchange and protein kinase C. An amiloride analog selectively inhibits Na+/H+ exchange. J Biol Chem. 1985 Jan 25;260(2):1155–1159. [PubMed] [Google Scholar]
  2. Boeynaems J. M., Galand N., Ketelbant P. Prostacyclin production by the deendothelialized rabbit aorta. J Clin Invest. 1985 Jul;76(1):7–14. doi: 10.1172/JCI111979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boeynaems J. M., Galand N. Stimulation of vascular prostacyclin synthesis by extracellular ADP and ATP. Biochem Biophys Res Commun. 1983 Apr 15;112(1):290–296. doi: 10.1016/0006-291x(83)91829-6. [DOI] [PubMed] [Google Scholar]
  4. Booyse F. M., Sedlak B. J., Rafelson M. E., Jr Culture of arterial endothelial cells: characterization and growth of bovine aortic cells. Thromb Diath Haemorrh. 1975 Dec 15;34(3):825–839. [PubMed] [Google Scholar]
  5. Chesterman C. N., Owe-Young R., Macpherson J., Krilis S. A. Substrate for endothelial prostacyclin production in the presence of platelets exposed to collagen is derived from the platelets rather than the endothelium. Blood. 1986 Jun;67(6):1744–1750. [PubMed] [Google Scholar]
  6. Cocks T. M., Little P. J., Angus J. A., Cragoe E. J., Jr Amiloride analogues cause endothelium-dependent relaxation in the canine coronary artery in vitro: possible role of Na+/Ca2+ exchange. Br J Pharmacol. 1988 Sep;95(1):67–76. doi: 10.1111/j.1476-5381.1988.tb16549.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cragoe E. J., Jr, Woltersdorf O. W., Jr, Bicking J. B., Kwong S. F., Jones J. H. Pyrazine diuretics. II. N-amidino-3-amino-5-substituted 6-halopyrazinecarboxamides. J Med Chem. 1967 Jan;10(1):66–75. doi: 10.1021/jm00313a014. [DOI] [PubMed] [Google Scholar]
  8. Demolle D., Lecomte M., Boeynaems J. M. Pattern of protein phosphorylation in aortic endothelial cells. Modulation by adenine nucleotides and bradykinin. J Biol Chem. 1988 Dec 5;263(34):18459–18465. [PubMed] [Google Scholar]
  9. Dieter P., Schulze-Specking A., Karck U., Decker K. Prostaglandin release but not superoxide production by rat Kupffer cells stimulated in vitro depends on Na+/H+ exchange. Eur J Biochem. 1987 Dec 30;170(1-2):201–206. doi: 10.1111/j.1432-1033.1987.tb13687.x. [DOI] [PubMed] [Google Scholar]
  10. FitzGerald G. A., Smith B., Pedersen A. K., Brash A. R. Increased prostacyclin biosynthesis in patients with severe atherosclerosis and platelet activation. N Engl J Med. 1984 Apr 26;310(17):1065–1068. doi: 10.1056/NEJM198404263101701. [DOI] [PubMed] [Google Scholar]
  11. Fitzgerald D. J., Roy L., Catella F., FitzGerald G. A. Platelet activation in unstable coronary disease. N Engl J Med. 1986 Oct 16;315(16):983–989. doi: 10.1056/NEJM198610163151602. [DOI] [PubMed] [Google Scholar]
  12. Hallam T. J., Pearson J. D. Exogenous ATP raises cytoplasmic free calcium in fura-2 loaded piglet aortic endothelial cells. FEBS Lett. 1986 Oct 20;207(1):95–99. doi: 10.1016/0014-5793(86)80019-9. [DOI] [PubMed] [Google Scholar]
  13. Jentsch T. J., Stahlknecht T. R., Hollwede H., Fischer D. G., Keller S. K., Wiederholt M. A bicarbonate-dependent process inhibitable by disulfonic stilbenes and a Na+/H+ exchange mediate 22Na+ uptake into cultured bovine corneal endothelium. J Biol Chem. 1985 Jan 25;260(2):795–801. [PubMed] [Google Scholar]
  14. Kaczorowski G. J., Barros F., Dethmers J. K., Trumble M. J., Cragoe E. J., Jr Inhibition of Na+/Ca2+ exchange in pituitary plasma membrane vesicles by analogues of amiloride. Biochemistry. 1985 Mar 12;24(6):1394–1403. doi: 10.1021/bi00327a017. [DOI] [PubMed] [Google Scholar]
  15. Kaila K., Voipio J. Postsynaptic fall in intracellular pH induced by GABA-activated bicarbonate conductance. Nature. 1987 Nov 12;330(6144):163–165. doi: 10.1038/330163a0. [DOI] [PubMed] [Google Scholar]
  16. Kitazono T., Takeshige K., Cragoe E. J., Jr, Minakami S. Intracellular pH changes of cultured bovine aortic endothelial cells in response to ATP addition. Biochem Biophys Res Commun. 1988 May 16;152(3):1304–1309. doi: 10.1016/s0006-291x(88)80427-3. [DOI] [PubMed] [Google Scholar]
  17. L'Allemain G., Paris S., Pouysségur J. Role of a Na+-dependent Cl-/HCO3- exchange in regulation of intracellular pH in fibroblasts. J Biol Chem. 1985 Apr 25;260(8):4877–4883. [PubMed] [Google Scholar]
  18. Lubin M., Cahn F., Coutermarsh B. A. Amiloride, protein synthesis, and activation of quiescent cells. J Cell Physiol. 1982 Nov;113(2):247–251. doi: 10.1002/jcp.1041130210. [DOI] [PubMed] [Google Scholar]
  19. Lucas C. A., Gillies R. J., Olson J. E., Giuliano K. A., Martinez R., Sneider J. M. Intracellular acidification inhibits the proliferative response in BALB/c-3T3 cells. J Cell Physiol. 1988 Jul;136(1):161–167. doi: 10.1002/jcp.1041360121. [DOI] [PubMed] [Google Scholar]
  20. Maclouf J. A radioimmunoassay for 6-keto-PGF1 alpha. Methods Enzymol. 1982;86:273–286. doi: 10.1016/0076-6879(82)86200-9. [DOI] [PubMed] [Google Scholar]
  21. Madshus I. H. Regulation of intracellular pH in eukaryotic cells. Biochem J. 1988 Feb 15;250(1):1–8. doi: 10.1042/bj2500001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Marcus A. J., Weksler B. B., Jaffe E. A., Broekman M. J. Synthesis of prostacyclin from platelet-derived endoperoxides by cultured human endothelial cells. J Clin Invest. 1980 Nov;66(5):979–986. doi: 10.1172/JCI109967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moolenaar W. H., Tsien R. Y., van der Saag P. T., de Laat S. W. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. doi: 10.1038/304645a0. [DOI] [PubMed] [Google Scholar]
  24. Needham L., Cusack N. J., Pearson J. D., Gordon J. L. Characteristics of the P2 purinoceptor that mediates prostacyclin production by pig aortic endothelial cells. Eur J Pharmacol. 1987 Feb 10;134(2):199–209. doi: 10.1016/0014-2999(87)90166-x. [DOI] [PubMed] [Google Scholar]
  25. Pearson J. D., Slakey L. L., Gordon J. L. Stimulation of prostaglandin production through purinoceptors on cultured porcine endothelial cells. Biochem J. 1983 Jul 15;214(1):273–276. doi: 10.1042/bj2140273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Perona R., Serrano R. Increased pH and tumorigenicity of fibroblasts expressing a yeast proton pump. Nature. 1988 Aug 4;334(6181):438–440. doi: 10.1038/334438a0. [DOI] [PubMed] [Google Scholar]
  27. Pirotton S., Raspe E., Demolle D., Erneux C., Boeynaems J. M. Involvement of inositol 1,4,5-trisphosphate and calcium in the action of adenine nucleotides on aortic endothelial cells. J Biol Chem. 1987 Dec 25;262(36):17461–17466. [PubMed] [Google Scholar]
  28. Pouysségur J., Sardet C., Franchi A., L'Allemain G., Paris S. A specific mutation abolishing Na+/H+ antiport activity in hamster fibroblasts precludes growth at neutral and acidic pH. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4833–4837. doi: 10.1073/pnas.81.15.4833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ritter J. M., Aksoy A., Cragoe E. J., Jr, Taylor G. W. Actions of amiloride analogues on prostacyclin synthesis by rat aortic rings. Br J Pharmacol. 1987 Dec;92(4):857–862. doi: 10.1111/j.1476-5381.1987.tb11391.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schafer A. I., Crawford D. D., Gimbrone M. A., Jr Unidirectional transfer of prostaglandin endoperoxides between platelets and endothelial cells. J Clin Invest. 1984 Apr;73(4):1105–1112. doi: 10.1172/JCI111296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schoeffter P., Miller R. C. Role of sodium-calcium exchange and effects of calcium entry blockers on endothelial-mediated responses in rat isolated aorta. Mol Pharmacol. 1986 Jul;30(1):53–57. [PubMed] [Google Scholar]
  32. Simchowitz L., Cragoe E. J., Jr Inhibition of chemotactic factor-activated Na+/H+ exchange in human neutrophils by analogues of amiloride: structure-activity relationships in the amiloride series. Mol Pharmacol. 1986 Aug;30(2):112–120. [PubMed] [Google Scholar]
  33. Soltoff S. P., Mandel L. J. Amiloride directly inhibits the Na,K-ATPase activity of rabbit kidney proximal tubules. Science. 1983 May 27;220(4600):957–958. doi: 10.1126/science.6302840. [DOI] [PubMed] [Google Scholar]
  34. Sweatt J. D., Connolly T. M., Cragoe E. J., Limbird L. E. Evidence that Na+/H+ exchange regulates receptor-mediated phospholipase A2 activation in human platelets. J Biol Chem. 1986 Jul 5;261(19):8667–8673. [PubMed] [Google Scholar]
  35. Sweatt J. D., Johnson S. L., Cragoe E. J., Limbird L. E. Inhibitors of Na+/H+ exchange block stimulus-provoked arachidonic acid release in human platelets. Selective effects on platelet activation by epinephrine, ADP, and lower concentrations of thrombin. J Biol Chem. 1985 Oct 25;260(24):12910–12919. [PubMed] [Google Scholar]
  36. Tang C. M., Presser F., Morad M. Amiloride selectively blocks the low threshold (T) calcium channel. Science. 1988 Apr 8;240(4849):213–215. doi: 10.1126/science.2451291. [DOI] [PubMed] [Google Scholar]
  37. Van Coevorden A., Boeynaems J. M. Physiological concentrations of ADP stimulate the release of prostacyclin from bovine aortic endothelial cells. Prostaglandins. 1984 Apr;27(4):615–626. doi: 10.1016/0090-6980(84)90097-2. [DOI] [PubMed] [Google Scholar]
  38. Vigne P., Frelin C., Cragoe E. J., Jr, Lazdunski M. Structure-activity relationships of amiloride and certain of its analogues in relation to the blockade of the Na+/H+ exchange system. Mol Pharmacol. 1984 Jan;25(1):131–136. [PubMed] [Google Scholar]
  39. Winquist R. J., Bunting P. B., Schofield T. L. Blockade of endothelium-dependent relaxation by the amiloride analog dichlorobenzamil: possible role of Na+/Ca++ exchange in the release of endothelium-derived relaxant factor. J Pharmacol Exp Ther. 1985 Dec;235(3):644–650. [PubMed] [Google Scholar]
  40. Yuli I., Oplatka A. Cytosolic acidification as an early transductory signal of human neutrophil chemotaxis. Science. 1987 Jan 16;235(4786):340–342. doi: 10.1126/science.3798116. [DOI] [PubMed] [Google Scholar]

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