Korean J Physiol Pharmacol.  2016 Jan;20(1):9-14. 10.4196/kjpp.2016.20.1.9.

cAMP induction by ouabain promotes endothelin-1 secretion via MAPK/ERK signaling in beating rabbit atria

• Peng LQ
• Li P
• Zhang QL
• Hong L
• Liu LP
• Cui X
• Cui BR

• KMID: 2150468
• DOI: http://doi.org/10.4196/kjpp.2016.20.1.9

Abstract

Adenosine 3',5'-cyclic monophosphate (cAMP) participates in the regulation of numerous cellular functions, including the Na(+)-K(+)-ATPase (sodium pump). Ouabain, used in the treatment of several heart diseases, is known to increase cAMP levels but its effects on the atrium are not understood. The aim of the present study was to examine the effect of ouabain on the regulation of atrial cAMP production and its roles in atrial endothelin-1 (ET-1) secretion in isolated perfused beating rabbit atria. Our results showed that ouabain (3.0 micromol/L) significantly increased atrial dynamics and cAMP levels during recovery period. The ouabain-increased atrial dynamics was blocked by KB-R7943 (3.0 micromol/L), an inhibitor for reverse mode of Na(+)-Ca(2+) exchangers (NCX), but did not by L-type Ca2+ channel blocker nifedipine (1.0 micromol/L) or protein kinase A (PKA) selective inhibitor H-89 (3.0 micromol/L). Ouabain also enhanced atrial intracellular cAMP production in response to forskolin and theophyline (100.0 micromol/L), an inhibitor of phosphodiesterase, potentiated the ouabain-induced increase in cAMP. Ouabain and 8-Bromo-cAMP (0.5 micromol/L) markedly increased atrial ET-1 secretion, which was blocked by H-89 and by PD98059 (30 micromol/L), an inhibitor of extracellular-signal-regulated kinase (ERK) without changing ouabain-induced atrial dynamics. Our results demonstrated that ouabain increases atrial cAMP levels and promotes atrial ET-1 secretion via the mitogen-activated protein kinase (MAPK)/ERK signaling pathway. These findings may explain the development of cardiac hypertrophy in response to digitalis-like compounds.

Keyword

Adenosine 3',5'-cyclic monophosphate (cAMP); Endothelin-1 (ET-1); Mitogen-activated protein kinase (MAPK); Ouabain

MeSH Terms

8-Bromo Cyclic Adenosine Monophosphate
Adenosine
Cardiomegaly
Colforsin
Cyclic AMP-Dependent Protein Kinases
Endothelin-1*
Heart Diseases
Nifedipine
Ouabain*
Phosphotransferases
Protein Kinases
8-Bromo Cyclic Adenosine Monophosphate
Adenosine
Colforsin
Cyclic AMP-Dependent Protein Kinases
Endothelin-1
Nifedipine
Ouabain
Phosphotransferases
Protein Kinases

Figure

• 

  Fig. 1 Effect of ouabain (3.0 µmol/L) on atrial stroke volume (A) and cAMP production (B).Data are presented as the means±SE (n=6). *p<0.05, **p<0.01, ***p<0.001, vs. the control period. ouabain (3.0 µmol/L)), please correct it as ouabain (3.0 µmol/L).

• 

  Fig. 2 Effects of forskolin (A) and theophyline (B) on ouabaininduced atrial adenylate cyclase activity in rabbit atria.Data are presented as the means±SE (n=6). **p<0.01, ***p<0.001 vs. control; #p<0.05, ♦♦p<0.01 vs. ouabain.

• 

  Fig. 3 Effect of H-89 (A), nifedipine (B), and KB-R7943 (C) on ouabain-induced atrial dynamics.Data are presented as the means±SE (n=6). **p<0.01, ***p<0.001 vs. Control period.

• 

  Fig. 4 Effects of ouabain (A) and 8-bromo-cAMP (B) on atrial ET-1 secretion in beating rabbit atria.Data are presented as the means±SE (n=6). *p<0.05, **p<0.01 vs. the control.

• 

  Fig. 5 Effects of PD98059 (A) and H-89 (B) on 8-Bromo-cAMP as well as ouabain (C) induced atrial ET-1 secretion in beating rabbit atria.*p<0.05, vs. control; **p<0.01 vs. the control; #p<0.01 vs. 8-Bromo-cAMP; ♦p<0.05 vs. ouabain.

Reference

1. Altamirano J, Li Y, DeSantiago J, Piacentino V 3rd, Houser SR, Bers DM. The inotropic effect of cardioactive glycosides in ventricular myocytes requires Na⁺-Ca²⁺ exchanger function. J Physiol. 2006; 575:845–854. PMID: 16825310.

2. Satoh H, Ginsburg KS, Qing K, Terada H, Hayashi H, Bers DM. KB-R7943 block of Ca²⁺ influx via Na⁺/Ca²⁺ exchange does not alter twitches or glycoside inotropy but prevents Ca²⁺ overload in rat ventricular myocytes. Circulation. 2000; 101:1441–1446. PMID: 10736290.

3. Saini HK, Dhalla NS. Sarcolemmal cation channels and exchangers modify the increase in intracellular calcium in cardiomyocytes on inhibiting Na⁺-K⁺-ATPase. Am J Physiol Heart Circ Physiol. 2007; 293:H169–H181. PMID: 17322410.

4. White CN, Liu CC, Garcia A, Hamilton EJ, Chia KK, Figtree GA, Rasmussen HH. Activation of cAMP-dependent signaling induces oxidative modification of the cardiac Na⁺-K⁺ pump and inhibits its activity. J Biol Chem. 2010; 285:13712–13720. PMID: 20194511.

5. Xie Z, Askari A. Na⁺/K⁺-ATPase as a signal transducer. Eur J Biochem. 2002; 269:2434–2439. PMID: 12027880.

6. Butcher RW, Sutherland EW. The effects of the catecholamines, adrenergic blocking agents, prostaglandin E1, and insulin on cyclie AMP levels in the rat epididymal fat pad in vitro. Ann N Y Acad Sci. 1967; 139:849–859. PMID: 4382732.

7. Almela P, Atucha NM, Milanés MV, Laorden ML. Cross-talk between protein kinase A and mitogen-activated protein kinases signalling in the adaptive changes observed during morphine withdrawal in the heart. J Pharmacol Exp Ther. 2009; 330:771–782. PMID: 19567779.

8. Liu LP, Hong L, Yu L, Li HY, Ding DZ, Jin SJ, Cui X. Ouabain stimulates atrial natriuretic peptide secretion via the endothelin-1/ET(B) receptor-mediated pathway in beating rabbit atria. Life Sci. 2012; 90:793–798. PMID: 22521291.

9. Baudouin-Legros M, Brouillard F, Tondelier D, Hinzpeter A, Edelman A. Effect of ouabain on CFTR gene expression in human Calu-3 cells. Am J Physiol Cell Physiol. 2003; 284:C620–C626. PMID: 12556359.

10. Bagrov AY, Fedorova OV, Austin-Lane JL, Dmitrieva RI, Anderson DE. Endogenous marinobufagenin-like immunoreactive factor and Na⁺, K⁺ ATPase inhibition during voluntary hypoventilation. Hypertension. 1995; 26:781–788. PMID: 7591018.

11. Mandal A, Delamere NA, Shahidullah M. Ouabain-induced stimulation of sodium-hydrogen exchange in rat optic nerve astrocytes. Am J Physiol Cell Physiol. 2008; 295:C100–C110. PMID: 18448627.

12. Westenfelder C, Sack E, Stuart E, Earnest W, Baranowski R, Kurtzman N. Effect of ouabain on basal and PTH-stimulated cAMP production in dog renal cortex (Abstract). Kidney Int. 1979; 16:844.

13. Mørk A, Geisler A. A comparative study on the effects of tetracyclines and lithium on the cyclic AMP second messenger system in rat brain. Prog Neuropsychopharmacol Biol Psychiatry. 1995; 19:157–169. PMID: 7708928.

14. Ishikawa S, Saito T. Effect of ouabain on cellular free calcium and cellular cyclic AMP production in response to arginine vasopressin in rat renal papillary collecting tubule cells in culture. J Endocrinol. 1989; 121:467–477. PMID: 2547011.

15. Cui X, Lee SJ, Kim SZ, Kim SH, Cho KW. Effects of pituitary adenylate cyclase activating polypeptide27 on cyclic AMP efflux and atrial dynamics in perfused beating atria. Eur J Pharmacol. 2000; 402:129–137. PMID: 10940366.

16. Cho KW, Kim SH, Hwang YH, Seul KH. Extracellular fluid translocation in perfused rabbit atria: implication in control of atrial natriuretic peptide secretion. J Physiol. 1993; 468:591–607. PMID: 8254526.

17. Bakker R, Groot JA. cAMP-mediated effects of ouabain and theophylline on paracellular ion selectivity. Am J Physiol. 1984; 246:G213–G217. PMID: 6320677.

18. Gagerman E, Hellman B, Täljedal IB. Effects of ouabain on insulin release, adenosine 3',5'-monophosphate and guanine 3',5'-monophosphate in pancreatic islets. Endocrinology. 1979; 104:1000–1002. PMID: 86435.

19. Cui X, Wen JF, Jin H, Li D, Jin JY, Kim SH, Kim SZ, Lee HS, Cho KW. Subtype-specific roles of cAMP phosphodiesterases in regulation of atrial natriuretic peptide release. Eur J Pharmacol. 2002; 451:295–302. PMID: 12242091.

20. Muller B, Lugnier C, Stoclet JC. Involvement of rolipram-sensitive cyclic AMP phosphodiesterase in the regulation of cardiac contraction. J Cardiovasc Pharmacol. 1990; 16:796–803. PMID: 1703603.

21. Shahid M, Nicholson CD. Comparison of cyclic nucleotide phosphodiesterase isoenzymes in rat and rabbit ventricular myocardium: positive inotropic and phosphodiesterase inhibitory effects of Org 30029, milrinone and rolipram. Naunyn Schmiedebergs Arch Pharmacol. 1990; 342:698–705. PMID: 1710786.

22. Bers DM, Ziolo MT. When is cAMP not cAMP? Effects of compartmentalization. Circ Res. 2001; 89:373–375. PMID: 11532895.

23. Hohl CM, Li QA. Compartmentation of cAMP in adult canine ventricular myocytes. Relation to single-cell free Ca²⁺ transients. Circ Res. 1991; 69:1369–1379. PMID: 1682066.