Enhanced Na+, K+-ATPase activity and endothelial modulation decrease phenylephrine-induced contraction in aorta from ouabain-treated normotensive and hypertensive rats

Ana Paula Davel; Gisele Kruger Couto; Camilla Ferreira Wenceslau; Emilia Cristina Peres; Fabiano Elias Xavier; Luciana Venturini Rossoni

doi:10.1515/hmbci-2013-0058

Article

Enhanced Na⁺, K⁺-ATPase activity and endothelial modulation decrease phenylephrine-induced contraction in aorta from ouabain-treated normotensive and hypertensive rats

Ana Paula Davel , Gisele Kruger Couto , Camilla Ferreira Wenceslau , Emilia Cristina Peres , Fabiano Elias Xavier and Luciana Venturini Rossoni

Published/Copyright: December 18, 2013

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From the journal Hormone Molecular Biology and Clinical Investigation Volume 18 Issue 2

Abstract

Aim: The purpose of this study was to compare the effect of long-term ouabain treatment on the vascular reactivity and Na⁺, K⁺-ATPase activity of a conductance artery from normotensive and hypertensive rats.

Methods: Male Wistar rats were treated with ouabain (~8.0 µg/day, subcutaneously) or vehicle for 5 and 20 weeks, and spontaneously hypertensive rats (SHRs) for 5 weeks. Vasoconstrictor response to phenylephrine (10⁻¹⁰ to 10⁻⁴ M) and relaxation curves to KCl (1–10 mM) were analyzed in thoracic aorta. The effects of endothelial removal, L-NAME (100 μM), and indomethacin (10 μM) were used to evaluate the endothelial, nitric oxide (NO), and cyclooxygenase (COX) modulation of phenylephrine response, respectively. Protein expression of endothelial and neuronal NO synthase (NOS) and COX-2 were also investigated.

Results: The phenylephrine-induced contraction was reduced, whereas the relaxation to KCl was enhanced in the aorta of ouabain-treated Wistar rats and SHRs. In both strains, endothelial modulation of α-adrenergic response was enhanced, related to an increased NO and reduced COX-derived vasoconstrictor factor modulation. Aortas from 20-week ouabain-treated Wistar rats showed reduced COX-2 and enhanced eNOS protein expression. In SHRs, 5-week ouabain treatment reduced COX-2 and increased nNOS protein expression.

Conclusions: The results suggest that long-term ouabain treatment reduces the α-adrenergic response of aorta from normotensive rats and SHRs, associated with an increase of NO synthesis, reduced COX-2-derived vasoconstrictor factors, and enhanced ouabain-sensitive Na⁺, K⁺-ATPase activity. These aortic mechanisms could be adjustments to the elevated blood pressure induced by ouabain, even in the presence of preexisting hypertension.

Keywords: aorta; cyclooxygenase-2; endothelium; Na+, K+-ATPase; ouabain

Corresponding author: Ana Paula Davel, Institute of Biology, Department of Structural and Functional Biology, Building J, Laboratory 04, State University of Campinas-UNICAMP, Rua Monteiro Lobato, 255, CEP 13083-862 Campinas, SP, Brazil, Phone: +55 19 35216189, Fax: +55 19 35216185, E-mail: anadavel@unicamp.br

Acknowledgments

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Desenvolvimento Cientíﬁco e Tecnológico (CNPq). F.E. Xavier and L.V. Rossoni are research fellows from CNPq.

References

1. Hamlyn JM, Blaustein MP, Bova S, DuCharme DW, Harris DW, Mandel F, Mathews WR, Ludens JH. Identification and characterization of a ouabain-like compound from human plasma. Proc Natl Acad Sci USA 1991;88:6259–63.10.1073/pnas.88.14.6259Search in Google Scholar PubMed PubMed Central

2. Manunta P, Rogowski AC, Hamilton BP, Hamlyn JM. Ouabain-induced hypertension in the rat: relationships among plasma and tissue ouabain and blood pressure. J Hypertens 1994;12:549–60.10.1097/00004872-199405000-00008Search in Google Scholar

3. Ludens JH, Clark MA, Robinson FG, DuCharme DW. Rat adrenal cortex is a source of a circulating ouabain-like compound. Hypertension 1992;19:721–4.10.1161/01.HYP.19.6.721Search in Google Scholar PubMed

4. Huang BS, Leenen FH. Brain renin-angiotensin system and ouabain-induced sympathetic hyperactivity and hypertension in Wistar rats. Hypertension 1999;34:107–12.10.1161/01.HYP.34.1.107Search in Google Scholar

5. Manunta P, Ferrandi M, Bianchi G, Hamlyn JM. Endogenous ouabain in cardiovascular function and disease. J Hypertens 2009;27:9–18.10.1097/HJH.0b013e32831cf2c6Search in Google Scholar PubMed

6. Blaustein MP, Leenen FH, Chen L, Golovina VA, Hamlyn JM, Pallone TL, Van Huysse JW, Zhang J, Wier WG. How NaCl raises blood pressure: a new paradigm for the pathogenesis of salt-dependent hypertension. Am J Physiol 2012;302:H1031–49.10.1152/ajpheart.00899.2011Search in Google Scholar PubMed PubMed Central

7. Hamlyn JM, Ringel R, Schaeffer J, Levinson PD, Hamilton BP, Kowearski AA, Blaustein MP. A circulating inhibitor of (Na+, K+) ATPase associated with essential hypertension. Nature 1982;300:650–2.10.1038/300650a0Search in Google Scholar PubMed

8. Rossoni LV, Salaices M, Marín J, Vassallo DV, Alonso MJ. Alterations in phenylephrine-induced contractions and the vascular expression of Na⁺,K⁺-ATPase in ouabain-induced hypertension. Br J Pharmacol 2002;135:771–81.10.1038/sj.bjp.0704501Search in Google Scholar PubMed PubMed Central

9. Rossoni LV, Salaices M, Miguel M, Briones AM, Barker LA, Vassallo DV, Alonso MJ. Ouabain-induced hypertension is accompanied by increases in endothelial vasodilator factors. Am J Physiol Heart Circ Physiol 2002;283:110–8.10.1152/ajpheart.00454.2002Search in Google Scholar PubMed

10. Xavier FE, Rossoni LV, Alonso MJ, Balfagón G, Vassallo DV, Salaices M. Ouabain induced hypertension alters the participation of endothelial factors in α-adrenergic responses differently in rat resistance and conductance mesenteric arteries. Br J Pharmacol 2004;143:215–25.10.1038/sj.bjp.0705919Search in Google Scholar PubMed PubMed Central

11. Xavier FE, Yogi A, Callera GE, Tostes RC, Alvarez Y, Salaices M, Alonso MJ, Rossoni LV. Contribution of the endothelin and renin-angiotensin systems to the vascular changes in rats chronically treated with ouabain. Br J Pharmacol 2004;143:794–802.10.1038/sj.bjp.0705994Search in Google Scholar PubMed PubMed Central

12. Hernanz R, Briones AM, Martín A, Beltrán AE, Tejerina T, Salaices M, Alonso MJ. Ouabain treatment increases nitric oxide bioavailability and decreases superoxide anion production in cerebral vessels. J Hypertens 2008;26:1944–54.10.1097/HJH.0b013e328308de55Search in Google Scholar PubMed

13. Briones AM, Padilha AS, Cogolludo AL, Alonso MJ, Vassallo DV, Pérez-Vizcaino F, Salaices M. Activation of BKCa channels by nitric oxide prevents coronary artery endothelial dysfunction in ouabain-induced hypertensive rats. J Hypertens 2009;27:83–91.10.1097/HJH.0b013e328317a7cfSearch in Google Scholar

14. Wenceslau CF, Davel AP, Xavier FE, Rossoni LV. Long-term ouabain treatment impairs vascular function in resistance arteries. J Vasc Res 2011;48:316–26.10.1159/000322576Search in Google Scholar PubMed

15. Pulgar VM, Jeffers AB, Rashad HM, Diz DI, Aileru AA. Increased constrictor tone induced by ouabain treatment in rats. J Cardiovasc Pharmacol 2013;62:174–83.10.1097/FJC.0b013e3182955d33Search in Google Scholar PubMed PubMed Central

16. Kimura K, Manunta P, Hamilton BP, Hamlyn JM. Different effects of in vivo ouabain and digoxin on renal artery function and blood pressure in the rat. Hypertens Res 2000;23:67–76.10.1291/hypres.23.Supplement_S67Search in Google Scholar

17. Linde CI, Antos LK, Golovina VA, Blaustein MP. Nanomolar ouabain increases NCX1 expression and enhances Ca²⁺ signaling in human arterial myocytes: a mechanism that links salt to increased vascular resistance? Am J Physiol Heart Circ Physiol 2012;303:H784–94.10.1152/ajpheart.00399.2012Search in Google Scholar PubMed PubMed Central

18. Cargnelli G, Trevisi L, Debetto P, Luciani S, Bova S. Effect of long-term ouabain treatment on contractile responses of rat aortae. J Cardiovasc Pharmacol 2000;35:538–42.10.1097/00005344-200004000-00004Search in Google Scholar PubMed

19. Songu-Mize E, Vassallo DV, Rashed SM, Varner KJ. Ouabain amplifies contractile responses to phenylephrine in rat tail arteries in hypertension. J Basic Clin Physiol Pharmacol 1995;6:309–19.10.1515/JBCPP.1995.6.3-4.309Search in Google Scholar PubMed

20. Vassallo DV, Songu-Mize E, Rossoni LV, Amaral SM. Effects of ouabain on vascular reactivity. Braz J Med Biol Res 1997;30:545–52.10.1590/S0100-879X1997000400016Search in Google Scholar

21. Rossoni LV, Pinto VD, Vassallo DV. Effects of small doses of ouabain on the arterial blood pressure of anesthetized hypertensive and normotensive rats. Braz J Med Biol Res 2001;34:1065–77.10.1590/S0100-879X2001000800014Search in Google Scholar

22. Padilha A, Rossoni LV, Xavier FE, Vassallo DV. Ouabain at nanomolar concentration promotes the synthesis and release of angiotensin II from the endothelium of the tail vascular bed of spontaneously hypertensive rats. J Cardiovasc Pharmacol 2004;44:372–80.10.1097/01.fjc.0000138165.96364.51Search in Google Scholar

23. Xavier FE, Davel AP, Fukuda LE, Rossoni LV. Chronic ouabain treatment exacerbates blood pressure elevation in spontaneously hypertensive rats: the role of vascular mechanisms. J Hypertens 2009;27:1233–42.10.1097/HJH.0b013e32832a391fSearch in Google Scholar

24. Gallo LC, Davel AP, Xavier FE, Rossoni LV. Time-dependent increases in ouabain-sensitive Na+, K+-ATPase activity in aortas from diabetic rats: the role of prostanoids and protein kinase C. Life Sci 2010;87:302–8.10.1016/j.lfs.2010.07.005Search in Google Scholar

25. Lüscher TF, Vanhoutte PM. Endothelium-dependent contractions to acetylcholine in the aorta of the spontaneously hypertensive rat. Hypertension 1986;8:344–8.10.1161/01.HYP.8.4.344Search in Google Scholar

26. Alvarez Y, Briones AM, Balfagón G, Alonso MJ, Salaices M. Hypertension increases the participation of vasoconstrictor prostanoids from cyclooxygenase-2 in phenylephrine responses. J Hypertens 2005;23:767–77.10.1097/01.hjh.0000163145.12707.63Search in Google Scholar

27. Chen GP, Li L, Yang Y, Fu M, Yao L, Wu T, Zhang XQ, Hu SJ. Chronic inhibition of farnesyl pyrophosphate synthase improves endothelial function in spontaneously hypertensive rats. Biochem Pharmacol 2010;80:1684–9.10.1016/j.bcp.2010.08.015Search in Google Scholar

28. Yang Q, Xue HM, Wong WT, Tian XY, Huang Y, Tsui SK, Ng PK, Wohlfart P, Li H, Xia N, Tobias S, Underwood MJ, He GW. AVE3085, an enhancer of endothelial nitric oxide synthase, restores endothelial function and reduces blood pressure in spontaneously hypertensive rats. Br J Pharmacol 2011;163:1078–85.10.1111/j.1476-5381.2011.01308.xSearch in Google Scholar

29. Nagakawa M, Takamatsu H, Toyoda T, Sawada S, Tsuji H, Ijichi H. Effect of inhibition of Na⁺,K⁺-ATPase on the prostacyclin generation of cultured human vascular endothelial cells. Life Sci 1987;40:351–7.10.1016/0024-3205(87)90136-6Search in Google Scholar

30. Alvarez Y, Pérez-Girón JV, Hernanz R, Briones AM, García-Redondo A, Beltrán A, Alonso MJ, Salaices M. Losartan reduces the increased participation of cyclooxygenase-2-derived products in vascular responses of hypertensive rats. J Pharmacol Exp Ther 2007;321:381–8.10.1124/jpet.106.115287Search in Google Scholar PubMed

31. Blaustein MP. Physiological effects of endogenous ouabain: control of intracellular Ca²⁺ stores and cell responsiveness. Am J Physiol 1993;264:C1367–87.10.1152/ajpcell.1993.264.6.C1367Search in Google Scholar

32. Marín J, Redondo J. Vascular sodium pump: endothelial modulation and alterations in some pathological processes and aging. Pharmacol Ther 1999;84:249–71.10.1016/S0163-7258(99)00037-6Search in Google Scholar

33. Kisters K, Krefting ER, Hausberg M, Kohnert KD, Honig A, Bettin D. Importance of decreased intracellular phosphate and magnesium concentrations and reduced ATPase activities in spontaneously hypertensive rats. Magnes Res 2000;13:183–8.Search in Google Scholar

34. Ponte A, Marín J, Arribas S, González R, Barrús MT, Salaices M, Sánchez-Ferrer CF. Endothelial modulation of ouabain-induced contraction and sodium pump activity in aortas of normotensive Wistar-Kyoto and spontaneously hypertensive rats. J Vasc Res 1996;33:164–74.10.1159/000159145Search in Google Scholar PubMed

35. Ponte A, Sánchez-Ferrer CF, Hernández C, Alonso MJ, Marín J. Effect of ageing and hypertension on endothelial modulation of ouabain-induced contraction and sodium pump activity in the rat aorta. J Hypertens 1996;14:705–12.10.1097/00004872-199606000-00005Search in Google Scholar PubMed

36. Liu J, Tian J, Haas M, Shapiro JI, Askari A, Xie Z. Ouabain interaction with cardiac Na⁺/K⁺-ATPase initiates signal cascades independent of changes in intracellular Na⁺ and Ca²⁺ concentrations. J Biol Chem 2000;275: 27838–44.10.1074/jbc.M002950200Search in Google Scholar PubMed

37. Xie Z, Cai T. Na⁺, K⁺-ATPase-mediated signal transduction: from protein interaction to cellular function. Mol Interv 2003;3:157–68.10.1124/mi.3.3.157Search in Google Scholar PubMed

38. Liu J, Xie Z. The sodium pump and cardiotonic steroids-induced signal transduction protein kinases and calcium-signaling microdomain in regulation of transporter trafficking. Biochim Biophys Acta 2010;1802:1237–45.10.1016/j.bbadis.2010.01.013Search in Google Scholar PubMed PubMed Central

39. Blanco G, Wallace DP. Novel role of ouabain as a cystogenic factor in autosomal dominant polycystic kidney disease. Am J Physiol Renal Physiol 2013;305:F797–812.10.1152/ajprenal.00248.2013Search in Google Scholar PubMed PubMed Central

40. Li Z, Xie Z. The Na/K-ATPase/Src complex and cardiotonic steroid-activated protein kinase cascades. Pflugers Arch 2009;457:635–44.10.1007/s00424-008-0470-0Search in Google Scholar PubMed

41. Eva A, Kirch U, Scheiner-Bobis G. Signaling pathways involving the sodium pump stimulate NO production in endothelial cells. Biochim Biophys Acta 2006;1758:1809–14.10.1016/j.bbamem.2006.09.006Search in Google Scholar PubMed

Received: 2013-9-30

Accepted: 2013-11-20

Published Online: 2013-12-18

Published in Print: 2014-5-1

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Articles in the same Issue

https://doi.org/10.1515/hmbci-2013-0058

Keywords for this article

aorta; cyclooxygenase-2; endothelium; Na+, K+-ATPase; ouabain