Basic Research in Cardiology

, Volume 101, Issue 2, pp 168–179 | Cite as

Post–conditioning reduces infarct size in the isolated rat heart: Role of coronary flow and pressure and the nitric oxide/cGMP pathway

  • C. Penna
  • S. Cappello
  • D. Mancardi
  • S. Raimondo
  • R. Rastaldo
  • D. Gattullo
  • G. Losano
  • P. PagliaroEmail author


We aimed to assess the role of the nitric oxide (NO)–cGMP pathway in cardioprotection by brief intermittent ischemias at the onset of reperfusion (i.e., post–conditioning (Post–con)). We also evaluated the role of coronary flow and pressure in Post–con. Rat isolated hearts perfused at constant– flow or –pressure underwent 30 min global ischemia and 120 min reperfusion. Post–con obtained with brief ischemias of different duration (modified, MPost–con) was compared with Post–con obtained with ischemias of identical duration (classical, C–Post–con) and with ischemic preconditioning (IP). Infarct size was evaluated using nitro–blue tetrazolium staining and lactate dehydrogenase (LDH) release. In the groups, NO synthase (NOS) or guanylyl–cyclase (GC) was inhibited with LNAME and ODQ, respectively. In the subgroups, the enzyme immunoassay technique was used to quantify cGMP release. In the constant–flow model, M–Post–con and C–Post–con were equally effective, but more effective than IP in reducing infarct size. The cardioprotection by M–Post–con was only blunted by the NOS–inhibitor, but was abolished by the GC–antagonist. Post–ischemic cGMP release was enhanced by MPost–con. In the constant–pressure model IP, M–Post–con and C–Post–con were equally effective in reducing infarct size. Post–con protocols were more effective in the constant–flow than in the constant–pressure model. In all groups, LDH release during reperfusion was proportional to infarct size. In conclusion, Post–con depends upon GC activation, which can be achieved by NOS–dependent and NOS–independent pathways. The benefits of M– and CPost–con are similar. However, protection by Post–con is greater in the constant–flow than in the constant–pressure model.

Key words

Ischemia cardioprotection cGMP nitric oxide reperfusion 


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  1. 1.
    Acar C, Partington MT, Buckberg GD (1990) Studies of controlled reperfusion after ischemia. XVII. Reperfusion conditions: controlled reperfusion through an internal mammary artery graft – a new technique emphasizing fixed pressure versus fixed flow. J Thorac Cardiovasc Surg 100:724–736PubMedGoogle Scholar
  2. 2.
    Amrani M, Chester AH, Jayakumar J, Schyns CJ, Yacoub MH (1995) L–arginine reverses low coronary reflow and enhances post–ischaemic recovery of cardiac mechanical function. Cardiovasc Res 30:200–204CrossRefPubMedGoogle Scholar
  3. 3.
    Depre C, Fierain L, Hue L (1997) Activation of nitric oxide synthase by ischaemia in the perfused heart Cardiovasc Res 33:82–87PubMedGoogle Scholar
  4. 4.
    Gaboury J, Woodman RC, Granger DN, Reinhardt P, Kubes P (1993) Nitric oxide prevents leukocyte adherence: role of superoxide. Am J Physiol Heart Circ Physiol 265:H862–H867Google Scholar
  5. 5.
    Galagudza M, Kurapeev D, Minasian S, Valen G, Vaage J (2004) Ischemic postconditioning: brief ischemia during reperfusion converts persistent ventricular fibrillation into regular rhythm. Eur J Cardiothorac Surg 25:1006–1010CrossRefPubMedGoogle Scholar
  6. 6.
    Giraldez RR, Panda A, Xia Y, Sanders SP, Zweier JL (1997) Decreased nitric–oxide synthase activity causes impaired endothelium– dependent relaxation in the postischemic heart. J Biol Chem 272:21420–21426CrossRefPubMedGoogle Scholar
  7. 7.
    Godber BL, Doel JJ, Sapkota GP, Blake DR, Stevens CR, Eisenthal R, Harrison R (2000) Reduction of nitrite to nitric oxide catalyzed by xanthine oxidoreductase. J Biol Chem 275:7757–7763CrossRefPubMedGoogle Scholar
  8. 8.
    Gross GJ (2000) The role of mitochondrial KATP channels in cardioprotection. Basic Res Cardiol 95:280–284CrossRefPubMedGoogle Scholar
  9. 9.
    Halkos ME, Kerendi F, Corvera JS, Wang NP, Kin H, Payne CS, Sun HY, Guyton RA, Vinten–Johansen J, Zhao ZQ (2004) Myocardial protection with postconditioning is not enhanced by ischemic preconditioning. Ann Thorac Surg 78:961–919CrossRefPubMedGoogle Scholar
  10. 10.
    Han H, Kaiser R, Hu K, Laser M, Ertl G, Bauersachs J (2003) Selective modulation of endogenous nitric oxide formation in ischemia/reperfusion injury in isolated rat hearts–effects on regional myocardial flow and enzyme release. Basic Res Cardiol 98:165–174PubMedGoogle Scholar
  11. 11.
    Hattori R, Otani H, Maulik N, Das DK (2002) Pharmacological preconditioning with resveratrol: role of nitric oxide. Am J Physiol Heart Circ Physiol 282:H1988–H1995PubMedGoogle Scholar
  12. 12.
    Heusch G (2004) Postconditioning: old wine in a new bottle? J Am Coll Cardiol 44:1111–1112CrossRefPubMedGoogle Scholar
  13. 13.
    Hori M, Kitakaze M, Sato H, Takashima S, Iwakura K, Inoue M, Kitabatake A, Kamada T (1991) Staged reperfusion attenuates myocardial stunning in dogs. Role of transient acidosis during early reperfusion. Circulation 84:2135–2145PubMedGoogle Scholar
  14. 14.
    Jones SP, Girod WG, Palazzo AJ, Granger DN, Grisham MB, Jourd'Heuil D, Huang PL, Lefer DJ (1999) Myocardial ischemiareperfusion injury is exacerbated in absence of endothelial cell nitric oxide synthase. Am J Physiol Heart Circ Physiol 276:H1567–H1573Google Scholar
  15. 15.
    Kin H, Zhao ZQ, Sun HY, Wang NP, Corvera JS, Halkos ME, Kerendi F, Guyton RA, Vinten–Johansen J (2004) Postconditioning attenuates myocardial ischemia–reperfusion injury by inhibiting events in the early minutes of reperfusion. Cardiovasc Res 62:74–85CrossRefPubMedGoogle Scholar
  16. 16.
    Kostic MM, Schrader J (1992) Role of nitric oxide in reactive hyperemia of the guinea pig heart. Circ Res 70:208–212PubMedGoogle Scholar
  17. 17.
    Kozlov AV, Staniek K, Nohl H (1999) Nitrite reductase activity is a novel function of mammalian mitochondria. FEBS Lett 454:127–130CrossRefPubMedGoogle Scholar
  18. 18.
    Li H, Samouilov A, Liu X, Zweier JL (2001) Characterization of the magnitude and kinetics of xanthine oxidase–catalyzed nitrite reduction. Evaluation of its role in nitric oxide generation in anoxic tissues. J Biol Chem 276:24482–24489PubMedGoogle Scholar
  19. 19.
    Ma XL, Gao F, Liu GL, Lopez BL, Christopher TA, Fukuto JM, Wink DA, Feelisch M (1999) Opposite effects of nitric oxide and nitroxyl on postischemic myocardial injury. Proc Natl Acad Sci USA 96:14617–14622PubMedGoogle Scholar
  20. 20.
    McLean PG, Perretti M, Ahluwalia A (1999) Inducible expression of the kinin B1 receptor in the endotoxemic heart: mechanisms of des–Arg9bradykinininduced coronary vasodilation. Br J Pharmacol 128:275–282CrossRefPubMedGoogle Scholar
  21. 21.
    Millar TM, Stevens CR, Benjamin N, Eisenthal R, Harrison R, Blake DR (1998) Xanthine oxidoreductase catalyses the reduction of nitrates and nitrite to nitric oxide under hypoxic conditions. FEBS Lett 427:225–228CrossRefPubMedGoogle Scholar
  22. 22.
    Mocanu MM, Baxter GF, Yue Y, Critz SD, Yellon DM (2000) The p38 MAPK inhibitor, SB203580, abrogates ischaemic preconditioning in rat heart but timing of administration is critical. Basic Res Cardiol 95:472–478CrossRefPubMedGoogle Scholar
  23. 23.
    Nohl H, Staniek K, Sobhian B, Bahrami S, Redl H, Kozlov AV (2000) Mitochondria recycle nitrite back to the bioregulator nitric monoxide. Acta Biochim Pol 47:913–921PubMedGoogle Scholar
  24. 24.
    Pabla R, Buda AJ, Flynn DM, Blesse SA, Shin AM, Curtis MJ, Lefer DJ (1996) Nitric oxide attenuates neutrophil–mediated myocardial contractile dysfunction after ischemia and reperfusion. Circ Res 78:65– 72PubMedGoogle Scholar
  25. 25.
    Pabla R, Buda AJ, Flynn DM, Salzberg DB, Lefer DJ (1995) Intracoronary nitric oxide improves postischemic coronary blood flow and myocardial contractile function. Am J Physiol Heart Circ Physiol 269:H1113–H1121Google Scholar
  26. 26.
    Pagliaro P, Mancardi D, Rastaldo R, Penna C, Gattullo D, Miranda KM, Feelisch M, Wink DA, Kass DA, Paolocci N (2003) Nitroxyl affords thiol–sensitive myocardial protective effects akin to early preconditioning. Free Radic Biol Med 34:33–43CrossRefPubMedGoogle Scholar
  27. 27.
    Pagliaro P, Rastaldo R, Penna C, Mancardi D, Cappello S, Losano GA (2004) Nitric oxide (NO)–cyclic guanosine monophosphate (cGMP) pathway is involved in ischemic postconditioning in the isolated rat heart. Circulation 110:III–136Google Scholar
  28. 28.
    Pantos C, Bescond–Jacquet A, Tzeis S, Paizis I, Mourouzis I, Moraitis P, Malliopoulou V, Politi ED, Karageorgiou H, Varonos D, Cokkinos DV (2005) Trimetazidine protects isolated rat hearts against ischemia–reperfusion injuryin an experimental timing –dependent manner. Basic Res Cardiol 100:154–160PubMedGoogle Scholar
  29. 29.
    Paolocci N, Ekelund UE, Isoda T, Ozaki M, Vandegaer K, Georgakopoulos D, Harrison RW, Kass DA, Hare JM (2000) cGMP–independent inotropic effects of nitric oxide and peroxynitrite donors: potential role for nitrosylation. Am J Physiol Heart Circ Physiol 279:H1982–H1988PubMedGoogle Scholar
  30. 30.
    Piper HM, Abdallah Y, Schafer C (2004) The first minutes of reperfusion: a window of opportunity for cardioprotection. Cardiovasc Res 61:365–371PubMedGoogle Scholar
  31. 31.
    Qin Q, Yang XM, Cui L, Critz SD, Cohen MV, Browner N, Lincoln TM, Downey JM (2004) Exogenous nitric oxide triggers preconditioning by a cGMPand mKATP–dependent mechanism. Am J Physiol Heart Circ Physiol. 287:H712–H718CrossRefPubMedGoogle Scholar
  32. 32.
    Rosenkranz AC, Hood SG, Woods RL, Dusting GJ, Ritchie RH (2002) Acute antihypertrophic actions of bradykinin in the rat heart: importance of cyclic GMP. Hypertension 40:498–503CrossRefPubMedGoogle Scholar
  33. 33.
    Sato H, Jordan JE, Zhao ZQ, Sarvotham SS, Vinten–Johansen J (1997) Gradual reperfusion reduces infarct size and endothelial injury but augments neutrophil accumulation. Ann Thorac Surg 64:1099–1107CrossRefPubMedGoogle Scholar
  34. 34.
    Schulz R, Kelm M, Heusch G (2004) Nitric oxide in myocardial ischemia/reperfusion injury. Cardiovasc Res 61:402–413CrossRefPubMedGoogle Scholar
  35. 35.
    Sun HY, Wang NP, Kerendi F, Halkos M, Kin H, Guyton RA, Vinten–Johansen J, Zhao ZQ (2005) Hypoxic postconditioning reduces cardiomyocyte loss by inhibiting ROS generation and intracellular Ca2+ overload. Am J Physiol Heart Circ Physiol 288:H1900–H1908PubMedGoogle Scholar
  36. 36.
    Thom SR, Fisher D, Zhang J, Bhopale VM, Ohnishi ST, Kotake Y, Ohnishi T, Buerk DG (2003) Stimulation of perivascular nitric oxide synthesis by oxygen. Am J Physiol Heart Circ Physiol 284:H1230–H1239PubMedGoogle Scholar
  37. 37.
    Tiravanti E, Samouilov A, Zweier JL (2004) Nitrosyl–heme complexes are formed in the ischemic heart: evidence of nitrite–derived nitric oxide formation, storage, and signaling in post–ischemic tissues. J Biol Chem 279:11065–11073CrossRefPubMedGoogle Scholar
  38. 38.
    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 95:230–232CrossRefPubMedGoogle Scholar
  39. 39.
    Valen G, Vaage J (2005) Pre– and postconditioning during cardiac surgery. Basic Res Cardiol 100:179–186CrossRefPubMedGoogle Scholar
  40. 40.
    Vinten–Johansen J, Zhao ZQ, Zatta AJ, Kin H, Halkos ME, Kerendi F (2005) Postconditioning A new link in nature's armor against myocardial ischemiareperfusion injury. Basic Res Cardiol 100:295–310PubMedGoogle Scholar
  41. 41.
    Wang P, Zweier JL (1996) Measurement of nitric oxide and peroxynitrite generation in the postischemic heart. J Biol Chem 271:29223–29230PubMedGoogle Scholar
  42. 42.
    Wink DA, Miranda KM, Katori T, Mancardi D, Thomas DD, Ridnour L, Espey MG, Feelisch M, Colton CA, Fukuto JM, Pagliaro P, Kass DA, Paolocci N (2003) Orthogonal properties of the redox siblings nitroxyl and nitric oxide in the cardiovascular system: a novel redox paradigm. Am J Physiol Heart Circ Physiol 285:H2264–H2276PubMedGoogle Scholar
  43. 43.
    Woditsch I, Schror K (1992) Prostacyclin rather than endogenous nitric oxide is a tissue protective factor in myocardial ischemia. Am J Physiol Heart Circ Physiol 263:H1390–H1396Google Scholar
  44. 44.
    Yang XM, Philipp S, Downey JM, Cohen MV. (2005) Postconditioning's protection is not dependent on circulating blood factors or cells but involves adenosine receptors and requires PI3–kinase and guanylyl cyclase activation. Basic Res Cardiol 100:57–63PubMedGoogle Scholar
  45. 45.
    Yang X–M, Proctor JB, Cui L, Krieg T, Downey JM, Cohen MV (2004) Multiple, brief coronary occlusions during early reperfusion protect rabbit hearts by targeting cell signalling pathways. J Am Coll Cardiol 44:1103–1110CrossRefPubMedGoogle Scholar
  46. 46.
    Zhang Z, Naughton D, Winyard PG, Benjamin N, Blake DR, Symons MC (1998) Generation of nitric oxide by a nitrite reductase activity of xanthine oxidase: a potential pathway for nitric oxide formation in the absence of nitric oxide synthase activity. Biochem Biophys Res Commun 249:767–772CrossRefPubMedGoogle Scholar
  47. 47.
    Zhao ZQ, Corvera JS, Halkos ME, Kerendi F, Wang NP, Guyton RA, Vinten– Johansen J (2003) Inhibition of myocardial injury by ischemic postconditioning during reperfusion: comparison with ischemic preconditioning. Am J Physiol Heart Circ Physiol 285:H579–H588PubMedGoogle Scholar
  48. 48.
    Zweier JL, Samouilov A, Kuppusamy P (1999) Non–enzymatic nitric oxide synthesis in biological systems. Biochim Biophys Acta 1411:250–262PubMedGoogle Scholar

Copyright information

© Steinkopff-Verlag 2005

Authors and Affiliations

  • C. Penna
    • 2
  • S. Cappello
    • 3
  • D. Mancardi
    • 2
  • S. Raimondo
    • 2
  • R. Rastaldo
    • 3
  • D. Gattullo
    • 2
  • G. Losano
    • 3
  • P. Pagliaro
    • 1
    Email author
  1. 1.Dipartimento di Scienze Cliniche e Biologiche Università di TorinoOspedale S. Luigi, Regione GonzoleOrbassano (TO)Italy
  2. 2.Dipartimento di Scienze Cliniche e Biologiche dell'Università di TorinoOrbassano (TO)Italy
  3. 3.Sezione di Fisiologia del Dipartimento di Neuroscienze dell'Università di TorinoOrbassano (TO)Italy

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