Real-Time Measurement of Nitric Oxide in Coronary Outflow during Transient Myocardial Ischemia and Reperfusion

  • Yoshiaki Fukuhiro
  • Seiichi Mochizuki
  • Masami Goto
  • Takashi Fujiwara
  • Taiji Murakami
  • Hiroshi Inada
  • Hisao Masaki
  • Ichiro Morita
  • Fumihiko Kajiya
Part of the Progress in Experimental Cardiology book series (PREC, volume 1)


To examine the kinetics and determinants of the production and release of nitric oxide (NO) from a heart during transient myocardial ischemia and reperfusion, we directly measured NO in the coronary efluent from isolated beating rat hearts during reperfusion following transient myocardial ischemia using a newly developed NO microelectrode. Isolated rat hearts were perfused with an oxygenated Krebs-Henseleit buffered solution and were subjected to one-minute or ten-minute global ischemia followed by reperfusion at 100 cmH2O. The time course of measured NO current during reperfusion showed a monophasic pattern in the case of one-minute ischemia but a biphasic pattern in the case of ten-minute ischemia. Immediately after the onset of reperfusion, coronary flow increased almost stepwise after one-minute ischemia and gradually after ten-minute ischemia. After one-minute ischemia? measured NO current first stayed at a relatively low level and then gradually increased (monophasic pattern). After ten-minute ischemia, following a transient peak, the measured NO current gradually increased (biphasic pattern). There was an excellent linear relationship between coronary flow rate and the calculated amount of NO during the second rise of NO release in the case often-minute ischemia. These data suggest that the time course of NO release from a heart during reperfusion is determined by the production of NO during ischemia, which is ischemic-duration dependent, and by the reperfusion-rate dependent mechanism.


Nitric Oxide Wall Shear Stress Calculated Amount Biphasic Pattern Calcium Ionophore A23187 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Moncada S, Palmer RM, Higgs EA. 1991. Nitric oxide: physiology, pathophysiology, and pharmacology. Phannacol Rev 43(2):109–142.Google Scholar
  2. 2.
    Kelm M, Schrader J. 1990. Control of coronary vascular tone by nitric oxide. Circ Res 66(6):1561–1575.PubMedGoogle Scholar
  3. 3.
    Hare JM, Colucci WS. 1995. Role of nitric oxide in the regulation of myocardial function. Prog Cardiovasc Dis 38(2):155–166.PubMedCrossRefGoogle Scholar
  4. 4.
    Kelly RA, Balligant JL, Smith TW. 1996. Nitric oxide and cardiac function. Circ Res 79(3):363–380.PubMedGoogle Scholar
  5. 5.
    Node K, Kitakaze M, Kosaka H, Komamura K, Minamino T, Inoue M, Tada M, Hori M, Kamada T. 1996. Increased release of NO during ischemia reduces myocardial contractility and improves metabolic dysfunction. Circulation 93(2):356–364.PubMedGoogle Scholar
  6. 6.
    Node K, Kitakaze M, Kosaka H, Komamura K, Minamino T, Tada M, Inoue M, Hori M, Kamada T. 1995. Plasma nitric oxide end products are increased in the ischemic canine heart. Biochem Biophys Res Commun 211(2):370–374.PubMedCrossRefGoogle Scholar
  7. 7.
    Zweier JL, Wang P, Kuppusamy P. 1995. Direct measurement of nitric oxide generation in the ischemic heart using electron paramagnetic resonance spectroscopy. J Biol Chem 270(1):304–307.PubMedCrossRefGoogle Scholar
  8. 8.
    Malinski T, Taha Z. 1992. Nitric oxide release from a single cell measured in situ by a porphyrinic-based microsensor. Nature 358:676–678.PubMedCrossRefGoogle Scholar
  9. 9.
    Shibuki K. 1990. An electrochemical microprobe for detecting nitric oxide release in brain tissue. Neurosci Res 9(1):69–76.PubMedCrossRefGoogle Scholar
  10. 10.
    Ichimori K, Ishida H, Fukahori M, Nakazawa H, Murakami E. 1994. Practical nitric oxide measurement employing a nitric oxide-selective electrode. Rev Sci Instrum 65(8):2714–2718.CrossRefGoogle Scholar
  11. 11.
    Tschudi MR, Mesaros S, Luscher TF, Malinski T. 1996. Direct in situ measurement of nitric oxide in mesenteric resistance arteries. Hypertension 27(1):32–35.PubMedGoogle Scholar
  12. 12.
    Vallance P, Patton S, Bhagat K, MacAllister R, Radomski M, Moncada S, Malinski T. 1995. Direct measurement of nitric oxide in human beings. Lancet 345:153–154.CrossRefGoogle Scholar
  13. 13.
    Mochizuki S, Goto M, Hirano K, Fukuhiro Y, Kajiya F. 1996. Direct measurement of nitric oxide in arterial wall using newly developed nitric oxide microsensor. Biomed Eng 8(4):18–23.Google Scholar
  14. 14.
    Hecker M, Mulsch A, Bassenge E, Busse R. 1993. Vasoconstriction and increased flow: two principal mechanisms of shear stress-dependent endothelial autacoid release. Am J Physiol 265(3, Pt 2):H828–H833.PubMedGoogle Scholar
  15. 15.
    Cowan CL, Cohen RA. 1991. Two mechanisms mediate relaxation by bradykinin of pig coronary artery: NO-dependent and independent responses. Am J Physiol 261(3, Pt 2):H830–H835.PubMedGoogle Scholar
  16. 16.
    Masini E, Bianchi S, Mugnai L, Gambassi F, Lupini M, Pistelli A, Mannaioni PF. 1991. The effect of nitric oxide generators on ischemia reperfusion injury and histamine release in isolated perfused guinea-pig heart. Agents Actions 33(1–2):53–56.PubMedCrossRefGoogle Scholar
  17. 17.
    Ungureanu LD, Balligand JL, Kelly RA, Smith TW. 1995. Myocardial contractile dysfunction in the systemic inflammatory response syndrome: role of a cytokine-inducible nitric oxide synthase in cardiac myocytes. J Mol Cell Cardiol 27(1):155–167.Google Scholar
  18. 18.
    Oddis CV, Finkel MS. 1995. Cytokine-stimulated nitric oxide production inhibits mitochondrial activity in cardiac myocytes. Biochem Biophys Res Commun 213(3):1002–1009.PubMedCrossRefGoogle Scholar
  19. 19.
    Kanai AJ, Strauss HC, Truskey GA, Grews AL, Crunfeld S, Malinski T. 1995. Shear stress induces ATP-independent transient nitric oxide release from vascular endothelial cells, measured directly with a porphyrinic microsensor. Circ Res 77(2):284–293.PubMedGoogle Scholar
  20. 20.
    Korenaga R, Ando J, Tsuboi H, Yang W, Sakuma I, Toyo-oka T, Kamiya A. 1994. Laminar flow stimulates ATP-and shear stress-dependent nitric oxide production in cultured bovine endothelial cells. Biochem Biophys Res Commun 198(1):213–219.PubMedCrossRefGoogle Scholar
  21. 21.
    Kuchan MJ, Frangos JA. 1994. Role of calcium and calmodulin in flow-induced nitric oxide production in endothelial cells. Am J Physiol 266(3, Pt 1):C628–C636.PubMedGoogle Scholar
  22. 22.
    Parratt JR, Vegh A, Papp JG. 1995. Bradykinin as an endogenous myocardial protective substance with particular reference to ischemic preconditioning—a brief review of the evidence. Can J Physiol Pharmacol 73(7):837–842.PubMedGoogle Scholar
  23. 23.
    Masini E, Giannella E, Bianchi S, Palmerani B, Pistelli A, Mannaioni PF. 1988. Histamine release in acute coronary occlusion—reperfusion in isolated guinea-pig heart. Agents Actions 23(3–4):266–269.PubMedCrossRefGoogle Scholar
  24. 24.
    Tani M, Neely JR. 1989. Role of intracellular Na+ in CA2+ overload and depressed recovery of ventricular hnction of reperfused ischemic rat hearts. Possible involvement of H+/Na+ and Na+/ Ca2+ exchange. Circ Res 65(4):1045–1056.PubMedGoogle Scholar
  25. 25.
    Marban E, Kitakaze M, Chacko VP, Pike MM. 1988. Ca2+ transients in perfused hearts revealed by gated 19F NMR spectroscopy. Circ Res 63(3):673–678.PubMedGoogle Scholar
  26. 26.
    Fleming I, Busse R. 1995. Control and consequences of endothelial nitric oxide formation. Adv Phamacol 34(187):187–206.CrossRefGoogle Scholar
  27. 27.
    Busse R, Mulsch A. 1990. Calcium-dependent nitric oxide synthesis in endothelial cytosol is mediated by calmodulin. FEBS Lett 265(1–2):133–136.PubMedCrossRefGoogle Scholar
  28. 28.
    Boulanger C, Schini VB, Moncada S, Vanhoutte PM. 1990. Stimulation of cyclic GMP production in cultured endothelial cells of the pig by bradykinin, adenosine diphosphate, calcium ionophore A23187 and nitric oxide. Br J Phamacol 101(1):152–156.Google Scholar
  29. 29.
    Heslinga JW, Allaart CP, Westerhof N. 1996. Intramyocardial pressure measurements in the isolated perfused papillary muscle of rat heart. Eur J Morphol 34(1):55–62.PubMedCrossRefGoogle Scholar
  30. 30.
    Corson MA, James NL, Lana SE, Nerem RM, Berk BC, Harrison DG. 1996. Phosphorylation of endothelial nitric oxide synthase in response to fluid shear stress. Circ Res 79(5):984–991.PubMedGoogle Scholar
  31. 31.
    Ayajiki K, Kindermann M, Hecker M, Fleming I, Busse R. 1996. Intracellular pH and tyrosine phosphorylation but not calcium determine shear stress-induced nitric oxide production in native endothelial cells. Circ Res 78(5):745–746.Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Yoshiaki Fukuhiro
    • 1
  • Seiichi Mochizuki
    • 1
  • Masami Goto
    • 1
  • Takashi Fujiwara
    • 1
  • Taiji Murakami
    • 1
  • Hiroshi Inada
    • 1
  • Hisao Masaki
    • 1
  • Ichiro Morita
    • 1
  • Fumihiko Kajiya
    • 1
  1. 1.Kawasaki Medical SchoolJapan

Personalised recommendations