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Uncoupled eNOS annihilates neuregulin-1β-induced cardioprotection: a novel mechanism in pharmacological postconditioning in myocardial infarction

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Abstract

Myocardial infarct size can be limited by pharmacological postconditioning (pPC) with cardioprotective agents. Cardioprotective effects of neuregulin-1β (NRG) via activation of protein kinase B (Akt) and downstream pathways like endothelial nitric oxide synthase (eNOS) have been postulated based on results from cell culture experiments. The purpose of this study was to investigate if eNOS may be involved in pPC with NRG. NRG application in an ex vivo mouse model (C57Bl6) of ischemia–reperfusion injury was analyzed. Unexpectedly, the infarct size increased when NRG was infused starting 5 min prior to reperfusion, even though protective Akt and GSK3β phosphorylation were enhanced. In eNOS deficient mice, however, NRG significantly reduced the infarct size. Co-infusion of NRG and l-arginine (Arg) lead to a reduction in infarct size in wild type animals. Electron paramagnetic resonance measurements revealed that NRG treatment prior to reperfusion leads to an enhanced release of reactive oxygen species compared to controls and this effect is blunted by co-infusion of Arg. This study documents the cardioprotective mechanisms of NRG signaling to be mediated by GSK3β inactivation. This is the first study to show that this protection fails in situations with dysfunctional eNOS. In eNOS deficient mice NRG exerts its protective effect via the GSK3β pathway, suggesting that the eNOS can limit cardioprotection. As dysfunctional eNOS has been described in cardiovascular risk factors like diabetes, hypertension, and hypercholesterolemia these findings can help to explain lack of postconditioning performance in models of cardiovascular co-morbidities.

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Abbreviations

Akt:

Protein kinase B

Arg:

l-Arginine

Cyt C :

Cytochrome C

eNOS:

Endothelial NO-synthase

ErbB-receptor:

Erythroblastosis virus B oncogene

EPR:

Electron paramagnetic resonance

GSK3β:

Glycogen synthase kinase 3β

iPC:

Ischemic postconditioning

LAD:

Left anterior descending artery

LVDP:

Left ventricular developed pressure

mPTP:

Mitochondrial permeability transition pore

NRG:

Neuregulin-1β

pPC:

Pharmacological postconditioning

RISK:

Reperfusion injury salvage kinase

ROS:

Reactive oxygen species

References

  1. Skyschally A, van CP, Iliodromitis EK, Schulz R, Kremastinos DT, Heusch G (2009) Ischemic postconditioning: experimental models and protocol algorithms. Basic Res Cardiol 104:469–483

    Article  PubMed  Google Scholar 

  2. Granfeldt A, Lefer DJ, Vinten-Johansen J (2009) Protective ischaemia in patients: preconditioning and postconditioning. Cardiovasc Res 83:234–246

    Article  PubMed  CAS  Google Scholar 

  3. Zhu M, Feng J, Lucchinetti E, Fischer G, Xu L, Pedrazzini T, Schaub MC, Zaugg M (2006) Ischemic postconditioning protects remodeled myocardium via the PI3K–PKB/Akt reperfusion injury salvage kinase pathway. Cardiovasc Res 72:152–162

    Article  PubMed  CAS  Google Scholar 

  4. Gomez L, Paillard M, Thibault H, Derumeaux G, Ovize M (2008) Inhibition of GSK3beta by postconditioning is required to prevent opening of the mitochondrial permeability transition pore during reperfusion. Circulation 117:2761–2768

    Article  PubMed  CAS  Google Scholar 

  5. Hedhli N, Huang Q, Kalinowski A, Palmeri M, Hu X, Russell RR, Russell KS (2011) Endothelium-derived neuregulin protects the heart against ischemic injury. Circulation 123:2254–2262

    Article  PubMed  CAS  Google Scholar 

  6. Sawyer DB, Zuppinger C, Miller TA, Eppenberger HM, Suter TM (2002) Modulation of anthracycline-induced myofibrillar disarray in rat ventricular myocytes by neuregulin-1beta and anti-erbB2: potential mechanism for trastuzumab-induced cardiotoxicity. Circulation 105:1551–1554

    Article  PubMed  CAS  Google Scholar 

  7. Timolati F, Ott D, Pentassuglia L, Giraud MN, Perriard JC, Suter TM, Zuppinger C (2006) Neuregulin-1 beta attenuates doxorubicin-induced alterations of excitation–contraction coupling and reduces oxidative stress in adult rat cardiomyocytes. J Mol Cell Cardiol 41:845–854

    Article  PubMed  CAS  Google Scholar 

  8. Bian Y, Sun M, Silver M, Ho KK, Marchionni MA, Caggiano AO, Stone JR, Amende I, Hampton TG, Morgan JP, Yan X (2009) Neuregulin-1 attenuated doxorubicin-induced decrease in cardiac troponins. Am J Physiol Heart Circ Physiol 297:H1974–H1983

    Article  PubMed  CAS  Google Scholar 

  9. Zhao YY, Sawyer DR, Baliga RR, Opel DJ, Han X, Marchionni MA, Kelly RA (1998) Neuregulins promote survival and growth of cardiac myocytes. Persistence of ErbB2 and ErbB4 expression in neonatal and adult ventricular myocytes. J Biol Chem 273:10261–10269

    Article  PubMed  CAS  Google Scholar 

  10. Hausenloy DJ, Yellon DM (2004) New directions for protecting the heart against ischaemia–reperfusion injury: targeting the reperfusion injury salvage kinase (RISK)-pathway. Cardiovasc Res 61:448–460

    Article  PubMed  CAS  Google Scholar 

  11. da Silva CG, Specht A, Wegiel B, Ferran C, Kaczmarek E (2009) Mechanism of purinergic activation of endothelial nitric oxide synthase in endothelial cells. Circulation 119:871–879

    Article  PubMed  Google Scholar 

  12. Hallstrom S, Gasser H, Neumayer C, Fugl A, Nanobashvili J, Jakubowski A, Huk I, Schlag G, Malinski T (2002) S-Nitroso human serum albumin treatment reduces ischemia/reperfusion injury in skeletal muscle via nitric oxide release. Circulation 105:3032–3038

    Article  PubMed  CAS  Google Scholar 

  13. Vasquez-Vivar J, Kalyanaraman B, Martasek P, Hogg N, Masters BS, Karoui H, Tordo P, Pritchard KA Jr (1998) Superoxide generation by endothelial nitric oxide synthase: the influence of cofactors. Proc Natl Acad Sci USA 95:9220–9225

    Article  PubMed  CAS  Google Scholar 

  14. Xia Y, Dawson VL, Dawson TM, Snyder SH, Zweier JL (1996) Nitric oxide synthase generates superoxide and nitric oxide in arginine-depleted cells leading to peroxynitrite-mediated cellular injury. Proc Natl Acad Sci USA 93:6770–6774

    Article  PubMed  CAS  Google Scholar 

  15. Jolly SR, Kane WJ, Bailie MB, Abrams GD, Lucchesi BR (1984) Canine myocardial reperfusion injury. Its reduction by the combined administration of superoxide dismutase and catalase. Circ Res 54:277–285

    Article  PubMed  CAS  Google Scholar 

  16. Godecke A, Decking UK, Ding Z, Hirchenhain J, Bidmon HJ, Godecke S, Schrader J (1998) Coronary hemodynamics in endothelial NO synthase knockout mice. Circ Res 82:186–194

    Article  PubMed  CAS  Google Scholar 

  17. Lange SA, Wolf B, Schober K, Wunderlich C, Marquetant R, Weinbrenner C, Strasser RH (2007) Chronic angiotensin II receptor blockade induces cardioprotection during ischemia by increased PKC-epsilon expression in the mouse heart. J Cardiovasc Pharmacol 49:46–55

    Article  PubMed  CAS  Google Scholar 

  18. Dikalov S, Skatchkov M, Fink B, Bassenge E (1997) Quantification of superoxide radicals and peroxynitrite in vascular cells using oxidation of sterically hindered hydroxylamines and electron spin resonance. Nitric Oxide 1:423–431

    Article  PubMed  CAS  Google Scholar 

  19. Paulus WJ, Serizawa T, Grossman W (1982) Altered left ventricular diastolic properties during pacing-induced ischemia in dogs with coronary stenoses. Potentiation by caffeine. Circ Res 50:218–227

    Article  PubMed  CAS  Google Scholar 

  20. Kaplan P, Matejovicova M, Herijgers P, Flameng W (2005) Effect of free radical scavengers on myocardial function and Na+, K+-ATPase activity in stunned rabbit myocardium. Scand Cardiovasc J 39:213–219

    Article  PubMed  CAS  Google Scholar 

  21. Khandoudi N, Percevault-Albadine J, Bril A (1998) Comparative effects of carvedilol and metoprolol on cardiac ischemia–reperfusion injury. J Cardiovasc Pharmacol 32:443–451

    Article  PubMed  CAS  Google Scholar 

  22. Shandelya SM, Kuppusamy P, Weisfeldt ML, Zweier JL (1993) Evaluation of the role of polymorphonuclear leukocytes on contractile function in myocardial reperfusion injury. Evidence for plasma-mediated leukocyte activation. Circulation 87:536–546

    Article  PubMed  CAS  Google Scholar 

  23. Liu X, Gu X, Li Z, Li X, Li H, Chang J, Chen P, Jin J, Xi B, Chen D, Lai D, Graham RM, Zhou M (2006) Neuregulin-1/erbB-activation improves cardiac function and survival in models of ischemic, dilated, and viral cardiomyopathy. J Am Coll Cardiol 48:1438–1447

    Article  PubMed  CAS  Google Scholar 

  24. Gao R, Zhang J, Cheng L, Wu X, Dong W, Yang X, Li T, Liu X, Xu Y, Li X, Zhou M (2010) A phase II, randomized, double-blind, multicenter, based on standard therapy, placebo-controlled study of the efficacy and safety of recombinant human neuregulin-1 in patients with chronic heart failure. J Am Coll Cardiol 55:1907–1914

    Article  PubMed  CAS  Google Scholar 

  25. Jabbour A, Hayward CS, Keogh AM, Kotlyar E, McCrohon JA, England JF, Amor R, Liu X, Li XY, Zhou MD, Graham RM, Macdonald PS (2011) Parenteral administration of recombinant human neuregulin-1 to patients with stable chronic heart failure produces favourable acute and chronic haemodynamic responses. Eur J Heart Fail 13:83–92

    Article  PubMed  CAS  Google Scholar 

  26. Lemmens K, Fransen P, Sys SU, Brutsaert DL, De Keulenaer GW (2004) Neuregulin-1 induces a negative inotropic effect in cardiac muscle: role of nitric oxide synthase. Circulation 109:324–326

    Article  PubMed  CAS  Google Scholar 

  27. Fukazawa R, Miller TA, Kuramochi Y, Frantz S, Kim YD, Marchionni MA, Kelly RA, Sawyer DB (2003) Neuregulin-1 protects ventricular myocytes from anthracycline-induced apoptosis via erbB4-dependent activation of PI3-kinase/Akt. J Mol Cell Cardiol 35:1473–1479

    Article  PubMed  CAS  Google Scholar 

  28. Kuramochi Y, Cote GM, Guo X, Lebrasseur NK, Cui L, Liao R, Sawyer DB (2004) Cardiac endothelial cells regulate reactive oxygen species-induced cardiomyocyte apoptosis through neuregulin-1beta/erbB4 signaling. J Biol Chem 279:51141–51147

    Article  PubMed  CAS  Google Scholar 

  29. Moss MB, Brunini TM, Soares DM, Novaes Malagris LE, Roberts NB, Ellory JC, Mann GE, Mendes Ribeiro AC (2004) Diminished l-arginine bioavailability in hypertension. Clin Sci (Lond) 107:391–397

    Article  CAS  Google Scholar 

  30. de Meirelles LR, Resende AC, Matsuura C, Salgado A, Pereira NR, Cascarelli PG, Mendes-Ribeiro AC, Brunini TM (2011) Platelet activation, oxidative stress and overexpression of inducible nitric oxide synthase in moderate heart failure. Clin Exp Pharmacol Physiol 38:705–710

    Article  PubMed  Google Scholar 

  31. Pribis JP, Zhu X, Vodovotz Y, Ochoa JB (2012) Systemic arginine depletion after a murine model of surgery or trauma. JPEN J Parenter Enteral Nutr 36:53–59

    Article  PubMed  CAS  Google Scholar 

  32. Mikulski D, Molski M (2010) Quantitative structure-antioxidant activity relationship of trans-resveratrol oligomers, trans-4,4′-dihydroxystilbene dimer, trans-resveratrol-3-O-glucuronide, glucosides: trans-piceid, cis-piceid, trans-astringin and trans-resveratrol-4′-O-beta-d-glucopyranoside. Eur J Med Chem 45:2366–2380

    Article  PubMed  CAS  Google Scholar 

  33. Gonon AT, Jung C, Yang J, Sjoquist PO, Pernow J (2011) The combination of l-arginine and ischaemic post-conditioning at the onset of reperfusion limits myocardial injury in the pig. Acta Physiol (Oxf) 201:219–226

    Article  CAS  Google Scholar 

  34. Jung C, Gonon AT, Sjoquist PO, Lundberg JO, Pernow J (2010) Arginase inhibition mediates cardioprotection during ischaemia–reperfusion. Cardiovasc Res 85:147–154

    Article  PubMed  CAS  Google Scholar 

  35. Wunderlich C, Schober K, Schmeisser A, Heerwagen C, Tausche AK, Steinbronn N, Brandt A, Kasper M, Schwencke C, Braun-Dullaeus RC, Strasser RH (2008) The adverse cardiopulmonary phenotype of caveolin-1 deficient mice is mediated by a dysfunctional endothelium. J Mol Cell Cardiol 44:938–947

    Article  PubMed  CAS  Google Scholar 

  36. Khong S, Andrews K, Huynh N, Venardos K, Aprico A, Michell D, Zarei M, Moe K, Dusting G, Kaye D, Chin-Dusting J (2012) Arginase II inhibition prevents nitrate tolerance. Br J Pharmacol 166:2015–2023

    Article  PubMed  CAS  Google Scholar 

  37. Guo Y, Li Q, Wu WJ, Tan W, Zhu X, Mu J, Bolli R (2008) Endothelial nitric oxide synthase is not necessary for the early phase of ischemic preconditioning in the mouse. J Mol Cell Cardiol 44:496–501

    Article  PubMed  CAS  Google Scholar 

  38. Ye Y, Lin Y, Manickavasagam S, Perez-Polo JR, Tieu BC, Birnbaum Y (2008) Pioglitazone protects the myocardium against ischemia–reperfusion injury in eNOS and iNOS knockout mice. Am J Physiol Heart Circ Physiol 295:H2436–H2446

    Article  PubMed  CAS  Google Scholar 

  39. Ye Y, Martinez JD, Perez-Polo RJ, Lin Y, Uretsky BF, Birnbaum Y (2008) The role of eNOS, iNOS, and NF-kappaB in upregulation and activation of cyclooxygenase-2 and infarct size reduction by atorvastatin. Am J Physiol Heart Circ Physiol 295:H343–H351

    Article  PubMed  CAS  Google Scholar 

  40. Bell RM, Yellon DM (2003) Bradykinin limits infarction when administered as an adjunct to reperfusion in mouse heart: the role of PI3K, Akt and eNOS. J Mol Cell Cardiol 35:185–193

    Article  PubMed  CAS  Google Scholar 

  41. Tamareille S, Ghaboura N, Treguer F, Khachman D, Croue A, Henrion D, Furber A, Prunier F (2009) Myocardial reperfusion injury management: erythropoietin compared with postconditioning. Am J Physiol Heart Circ Physiol 297:H2035–H2043

    Article  PubMed  CAS  Google Scholar 

  42. Honisch A, Theuring N, Ebner B, Wagner C, Strasser RH, Weinbrenner C (2010) Postconditioning with levosimendan reduces the infarct size involving the PI3K pathway and KATP-channel activation but is independent of PDE-III inhibition. Basic Res Cardiol 105:155–167

    Article  PubMed  Google Scholar 

  43. Yin Z, Gao H, Wang H, Li L, Di C, Luan R, Tao L (2009) Ischaemic post-conditioning protects both adult and aged Sprague-Dawley rat heart from ischaemia–reperfusion injury through the phosphatidylinositol 3-kinase-AKT and glycogen synthase kinase-3beta pathways. Clin Exp Pharmacol Physiol 36:756–763

    Article  PubMed  CAS  Google Scholar 

  44. Miura T, Miki T (2009) GSK-3beta, a therapeutic target for cardiomyocyte protection. Circ J 73:1184–1192

    Article  PubMed  CAS  Google Scholar 

  45. Tsang A, Hausenloy DJ, Mocanu MM, Yellon DM (2004) Postconditioning: a form of “modified reperfusion” protects the myocardium by activating the phosphatidylinositol 3-kinase-Akt pathway. Circ Res 95:230–232

    Article  PubMed  CAS  Google Scholar 

  46. Sumi S, Kobayashi H, Yasuda S, Iwasa M, Yamaki T, Yamada Y, Ushikoshi H, Hattori A, Aoyama T, Nishigaki K, Takemura G, Minatoguchi S (2010) Postconditioning effect of granulocyte colony-stimulating factor is mediated through activation of risk pathway and opening of the mitochondrial KATP channels. Am J Physiol Heart Circ Physiol 299:H1174–H1182

    Article  PubMed  CAS  Google Scholar 

  47. Heitzer T, Brockhoff C, Mayer B, Warnholtz A, Mollnau H, Henne S, Meinertz T, Munzel T (2000) Tetrahydrobiopterin improves endothelium-dependent vasodilation in chronic smokers: evidence for a dysfunctional nitric oxide synthase. Circ Res 86:E36–E41

    Article  PubMed  CAS  Google Scholar 

  48. Heitzer T, Krohn K, Albers S, Meinertz T (2000) Tetrahydrobiopterin improves endothelium-dependent vasodilation by increasing nitric oxide activity in patients with type II diabetes mellitus. Diabetologia 43:1435–1438

    Article  PubMed  CAS  Google Scholar 

  49. Higashi Y, Sasaki S, Nakagawa K, Fukuda Y, Matsuura H, Oshima T, Chayama K (2002) Tetrahydrobiopterin enhances forearm vascular response to acetylcholine in both normotensive and hypertensive individuals. Am J Hypertens 15:326–332

    Article  PubMed  CAS  Google Scholar 

  50. Stroes E, Kastelein J, Cosentino F, Erkelens W, Wever R, Koomans H, Luscher T, Rabelink T (1997) Tetrahydrobiopterin restores endothelial function in hypercholesterolemia. J Clin Invest 99:41–46

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Janet Lehmann and Anita Männel for excellent technical assistance. The study was supported by the research prize of the Dresdener Herz-Kreislauf Tage awarded to Bernd Ebner.

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Author notes

  1. Annette Ebner

    Present address: Laboratory of Experimental and Molecular Cardiology, University of Technology Dresden, Dresden, Germany

  2. Stefan A. Lange

    Present address: Klinikum Worms, Gabriel-von-Seidl-Str. 81, 67550, Worms, Germany

  3. Rüdiger C. Braun-Dullaeus

    Present address: Department of Cardiology, Angiology and Pneumology, Otto-von-Guericke-Universität Magdeburg, Leipziger Str. 44, 39120, Magdeburg, Germany

  4. Christof Weinbrenner

    Present address: Medizinische Klinik I, Klinikum Hanau, Leimenstr. 20, 63450, Hanau, Germany

Authors and Affiliations

  1. Department of Medicine/Cardiology, Heart Center Dresden, University Hospital, University of Technology Dresden, Dresden, Germany

    Bernd Ebner, Stefan A. Lange, Thomas Eckert, Clementine Wischniowski, Rüdiger C. Braun-Dullaeus, Christof Weinbrenner, Carsten Wunderlich, Gregor Simonis & Ruth H. Strasser

  2. Herzzentrum Dresden, Fetscherstr. 76, 01307, Dresden, Germany

    Bernd Ebner

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Correspondence to Bernd Ebner.

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Bernd Ebner and Stefan A. Lange contributed equally to this study.

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Ebner, B., Lange, S.A., Eckert, T. et al. Uncoupled eNOS annihilates neuregulin-1β-induced cardioprotection: a novel mechanism in pharmacological postconditioning in myocardial infarction. Mol Cell Biochem 373, 115–123 (2013). https://doi.org/10.1007/s11010-012-1480-y

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