Cardiovascular Drugs and Therapy

, Volume 24, Issue 2, pp 95–106 | Cite as

Hypoxia Inducible Factor-1 Protects Against Nitrate Tolerance and Stunning in Rabbit Cardiac Myocytes

  • Tao Tan
  • Harvey R. WeissEmail author



We tested whether upregulation of hypoxia inducible factor-1 (HIF-1) would restore the blunted effects of natriuretic peptides and nitric oxide caused by chronic nitrate exposure and stunning in cardiac myocytes.


HIF-1α was increased with deferoxamine (150 mg/kg for 2 days). Nitrate tolerance was induced by a chronic nitroglycerin patch (0.3 mg/h for 5 days). We used freshly isolated rabbit ventricular myocytes. Half the myocytes were subjected to simulated ischemia [15 min 95% N2-5% CO2] and reperfusion [reoxygenation] to produce stunning. Cell function was measured utilizing a video-edge detector. Shortening was examined at baseline and after brain natriuretic peptide (BNP, 10−8, 10−7 M) or S-nitroso-N-acetyl-penicillamine (SNAP, 10−6, 10−5 M) followed by KT5823 (cyclic GMP protein kinase inhibitor, 10−6 M). We also measured cyclic GMP protein kinase protein levels and kinase activity.


In control, BNP (−29%) reduced percent shortening, while KT5823 partially restored function. Deferoxamine treated control myocytes responded similarly. In patched nonstunned myocytes, BNP (−12%) reduced shortening less and KT5823 did not increase function. However, deferoxamine restored the blunted effects of BNP (−21%) and KT5823. In stunned myocytes, BNP (−11%) reduced shortening less and KT5823 did not affect function. Deferoxamine increased the effects of BNP (−27%) and KT5823 in stunning. Patch combined with stunning also similarly blunted the effects of BNP (−12%) and KT5823. Deferoxamine improved the effects of BNP (−22%) and KT5823. Similar results were observed after SNAP. Stunning reduced cyclic GMP protein kinase activity and deferoxamine restored activity. Deferoxamine had no effect on kinase activity in nitrate tolerance.


We found that upregulation of HIF-1 could protect isolated cardiac myocytes against nitrate tolerance through a cyclic GMP protein kinase-independent mechanism and through a kinase-dependent mechanism in stunning.

Key words

Cardiac myocytes Hypoxia inducible factor-1 Nitrate tolerance Myocardial stunning Cyclic GMP signaling 


  1. 1.
    Murad F. Cellular signaling with nitric oxide and cyclic GMP. Braz J Med Biol Res. 1999;32:1317–27.PubMedGoogle Scholar
  2. 2.
    Thadani U. Nitrate tolerance, rebound, and their clinical relevance in stable angina pectoris, unstable angina, and heart failure. Cardiovasc Drugs Ther. 1997;10:735–42.CrossRefPubMedGoogle Scholar
  3. 3.
    Shah AM, MacCarthy PA. Paracrine and autocrine effects of nitric oxide on myocardial function. Pharmacol Ther. 2000;86:49–86.CrossRefPubMedGoogle Scholar
  4. 4.
    Tan T, Zhang Q, Anyadike C, Scholz PM, Weiss HR. Chronic nitrates blunt the effects of not only nitric oxide but also natriuretic peptides in cardiac myocytes. Pharmacol Res. 2007;56:49–55.CrossRefPubMedGoogle Scholar
  5. 5.
    Munzel T, Daiber A, Mulsch A. Explaining the phenomenon of nitrate tolerance. Circ Res. 2005;97:618–28.CrossRefPubMedGoogle Scholar
  6. 6.
    Heyndrickx GR. Myocardial stunning: an experimental act with a large clinical audience. Arch Mal Coeur Vaiss. 2003;96:665–70.PubMedGoogle Scholar
  7. 7.
    Anyadike C, Scholz PM, Zhang Q, Katz E, Weiss HR. Brain natriuretic peptide reverses the effects of myocardial stunning in rabbit myocardium. Pharmacology. 2007;80:40–8.CrossRefPubMedGoogle Scholar
  8. 8.
    Zhang Q, Lazar M, Yan L, et al. Cyclic GMP reduces myocardial stunning through non-cyclic GMP protein kinase mechanisms. J Cardiovasc Pharmacol. 2004;44:235–43.CrossRefPubMedGoogle Scholar
  9. 9.
    Dery MA, Michaud MD, Richard DE. Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int J Biochem Cell Biol. 2005;37:535–40.CrossRefPubMedGoogle Scholar
  10. 10.
    Poellinger L, Johnson RS. HIF-1 and hypoxic response: the plot thickens. Curr Opin Genet Dev. 2004;14:81–5.CrossRefPubMedGoogle Scholar
  11. 11.
    Semenza GL. O2-regulated gene expression: transcriptional control of cardiorespiratory physiology by HIF-1. J Appl Physiol. 2004;96:1173–7. discussion 0–2.CrossRefPubMedGoogle Scholar
  12. 12.
    Luciano JA, Tan T, Zhang Q, Huang E, Scholz P, Weiss HR. Hypoxia inducible factor-1 improves the actions of nitric oxide and natriuretic peptides after simulated ischemia-reperfusion. Cell Physiol Biochem. 2008;21:421–8.CrossRefPubMedGoogle Scholar
  13. 13.
    Su J, Zhang Q, Moalem J, Tse J, Scholz PM, Weiss HR. Functional effects of C-type natriuretic peptide and nitric oxide are attenuated in hypertrophic myocytes from pressure-overloaded mouse hearts. Am J Physiol Heart Circ Physiol. 2005;288:H1367–73.CrossRefPubMedGoogle Scholar
  14. 14.
    Zhu H, Cai C, Chen J. Suppression of P-glycoprotein gene expression in Hs578T/Dox by the overexpression of caveolin-1. FEBS letters. 2004;576:369–74.CrossRefPubMedGoogle Scholar
  15. 15.
    Ziolo MT, Kohr MJ, Wang H. Nitric oxide signaling and the regulation of myocardial function. J Mol Cell Cardiol. 2008;45:625–32.CrossRefPubMedGoogle Scholar
  16. 16.
    Yan L, Zhang Q, Scholz PM, Weiss HR. Cyclic GMP protein kinase activity is reduced in thyroxine-induced hypertrophic cardiac myocytes. Clin Exp Pharmacol Physiol. 2003;30:943–50.CrossRefPubMedGoogle Scholar
  17. 17.
    Francis SH, Blount MA, Zoraghi R, Corbin JD. Molecular properties of mammalian proteins that interact with cGMP: protein kinases, cation channels, phosphodiesterases, and multi-drug anion transporters. Front Biosci. 2005;10:2097–117.CrossRefPubMedGoogle Scholar
  18. 18.
    Kojda G, Kottenberg K. Regulation of basal myocardial function by NO. Cardiovasc Res. 1999;41:514–23.CrossRefPubMedGoogle Scholar
  19. 19.
    Klemenska E, Beresewicz A. Bioactivation of organic nitrates and the mechanism of nitrate tolerance. Cardiol J. 2009;16:11–9.PubMedGoogle Scholar
  20. 20.
    Gori T, Parker JD. Nitrate tolerance: a unifying hypothesis. Circulation. 2002;106:2510–3.CrossRefPubMedGoogle Scholar
  21. 21.
    Mulsch A, Oelze M, Kloss S, et al. Effects of in vivo nitroglycerin treatment on activity and expression of the guanylyl cyclase and cGMP-dependent protein kinase and their downstream target vasodilator-stimulated phosphoprotein in aorta. Circulation. 2001;103:2188–94.PubMedGoogle Scholar
  22. 22.
    Sage PR, de la Lande IS, Stafford I, et al. Nitroglycerin tolerance in human vessels: evidence for impaired nitroglycerin bioconversion. Circulation. 2000;102:2810–5.PubMedGoogle Scholar
  23. 23.
    Warnholtz A, Tsilimingas N, Wendt M, Munzel T. Mechanisms underlying nitrate-induced endothelial dysfunction: insight from experimental and clinical studies. Heart Fail Rev. 2002;7:335–45.CrossRefPubMedGoogle Scholar
  24. 24.
    Barnes E, Khan MA. Myocardial stunning in man. Heart Fail Rev. 2003;8:155–60.CrossRefPubMedGoogle Scholar
  25. 25.
    Gowda RM, Khan IA, Vasavada BC, Sacchi TJ. Reversible myocardial dysfunction: basics and evaluation. Int J Cardiol. 2004;97:349–53.CrossRefPubMedGoogle Scholar
  26. 26.
    Weiss HR, Gandhi A, Scholz PM. Negative effect of nitric oxide on shortening-frequency relationship in cardiac myocytes is diminished after simulated ischemia-reperfusion. Basic Res Cardiol. 2003;98:311–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Hannan RL, John MC, Kouretas PC, Hack BD, Matherne GP, Laubach VE. Deletion of endothelial nitric oxide synthase exacerbates myocardial stunning in an isolated mouse heart model. J Surg Res. 2000;93:127–32.CrossRefPubMedGoogle Scholar
  28. 28.
    Brune B, Zhou J. Nitric oxide and superoxide: interference with hypoxic signaling. Cardiovasc Res. 2007;75:275–82.CrossRefPubMedGoogle Scholar
  29. 29.
    Shrivastava K, Shukla D, Bansal A, Sairam M, Banerjee PK, Ilavazhagan G. Neuroprotective effect of cobalt chloride on hypobaric hypoxia-induced oxidative stress. Neurochem Int. 2008;52:368–75.CrossRefPubMedGoogle Scholar
  30. 30.
    Kirito K, Hu Y, Komatsu N. HIF-1 prevents the overproduction of mitochondrial ROS after cytokine stimulation through induction of PDK-1. Cell Cycle. 2009;8:2844–9.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Heart and Brain Circulation Laboratory, Department of Physiology and BiophysicsUniversity of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical SchoolPiscatawayUSA

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