Microvascular Oxygenation and Oxidative Stress During Postischemic Reperfusion

PO2, ROS, and NO during reperfusion
  • Silvia Bertuglia
  • Andrea Giusti
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 566)


Increased formation of ROS on reperfusion after ischemia underlies ischemia reperfusion (I/R) damage. We measured, in real time, both oxygen tension in microvessels and tissue and oxidant stress during postischemic reperfusion in hamster cheek pouch microcirculation. We measured PO2 by using phosphorescence quenching microscopy and oxygen radical species (ROS) production in the systemic blood. We evaluated the effects of a NOS inhibitor (L-NMMA) and superoxide dismutase (SOD) on the oxidative stress during reperfusion. Microvascular injury was assessed by measuring diameter change, the perfused capillary length (PCL), and leukocyte adhesion.

Our findings demonstrate that early reperfusion is characterized by low concentration of oxygen linked to increased production of ROS. After this initial transience in arterioles, the oxygen tension and production of ROS return to normal after reperfusion, while the blood flow and capillary perfusion decrease. The early increased ROS production, in turn, may impair oxygen consumption by endothelial cells, thus further promoting activation of oxygen to ROS. This event is substantiated by the finding that treatment with SOD maintains ROS at normal levels, which, in turn, should be effective to increase the production of endothelial NO. Conversely, a decrease in NO levels led to decreased ROS production during early reperfusion, which increased later during reperfusion, ultimately causing vasoconstriction and greatly increasing venular leukocyte adhesion on postcapillary venules during hypoxic conditions. Therefore, low-flow hypoxia is primarily responsible for vascular endothelial damage during reperfusion through changes in ROS and NO production.


Reactive Oxygen Species Nitric Oxide Mean Arterial Blood Pressure Early Reperfusion Cheek Pouch 
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  1. 1.
    B. Haliwell, and J. M. C. Gutteridge, Free radicals in biology and medicine, 3rd ed. Oxford University Press (1993).Google Scholar
  2. 2.
    S. Bertuglia, and A. Giusti, Microvascular oxygenation, oxidative stress, nitric oxide suppression and superoxide dismutase during postischemic reperfusion, Am. J. Physiol. 278, H1064–1071 (2003).Google Scholar
  3. 3.
    J. S. Beckman, and W. H. Koppenol, Nitric oxide, superoxide and peroxynitrite: the good, the bad and the ugly, Am. J. Physiol. 271, C1424–1437 (1996).PubMedGoogle Scholar
  4. 4.
    J. S. Beckman, T.W. J. Beckman, P. Chen, P. A. Marshall, and B. A. Freeman, Apparent hydroxyl radical production by peroxinitrite, implications for endothelial injury from NO and superoxide, Proc. Nat. Acad. Sci. USA 87, 1620–1624 (1990).PubMedCrossRefGoogle Scholar
  5. 5.
    L. Kenneth, C. J. Giovanelli, and S. Kaufman, Characteristics of the NO synthase-catalyzed conversion of arginine to N-hydroxyarginine, the first oxygenation step in the enzymic synthesis of NO, J. Biol. Chem. 270,1721–1728(1995).CrossRefGoogle Scholar
  6. 6.
    M. T. Gladwin, J. R. Lancaster, B. A. Freeman, and A. N. Schechter, NO’s reactions with hemoglobin a view through the SNO-storm, Nature Med. 9, 496–500 (2003).PubMedCrossRefGoogle Scholar
  7. 7.
    S. Bertuglia, A. Giusti, S. Fedele, and E. Picano, Glucose-insulin-potassium treatment in combination with dipyridamole inhibits ischemia-reperfusion-induced damage, Diabetologia 44, 2165–2170 (2001).PubMedCrossRefGoogle Scholar
  8. 8.
    A. Golub, A. S. Popel, L. Zheng, and R. N. Pittman, Analysis of phosphorescence in heterogeneous systems using distributions of quencher concentration, Biophys. J. 73, 452–465 (1999).CrossRefGoogle Scholar
  9. 9.
    M. Intaglietta, P. C. Johnson, and R. M. Winslow, Microvascular and tissue oxygen distribution, Cardiovasc. Res. 32, 632–643 (1996).PubMedCrossRefGoogle Scholar
  10. 10.
    T. Yamamoto, and R. J. Bing, NO donors, Proc. Soc. Exp. Biol. Med. 225, 200–206 (2000).PubMedCrossRefGoogle Scholar
  11. 11.
    A. S. Popel, R. N. Pittman, and M. L. Ellsworth, Rate of oxygen loss from arterioles is an order of magnitude higher than expected, Am. J. Physiol. 256, H921–924 (1989).PubMedGoogle Scholar
  12. 12.
    P. S. Tsao, and A. M. Lefer, Time course and mechanism of endothelial dysfunction in isolated ischemic and hypoxic perfused rat hearts, Am. J. Physiol. 259, H1660–1666 (1990).PubMedGoogle Scholar
  13. 13.
    A. R. Whorton, D. B. Simonds, and C. A. Piantadosi, Regulation of NO synthesis by oxygen in vascular endothelial cells, Am. J. Physiol. 272, L1161–1166, (1997).PubMedGoogle Scholar
  14. 14.
    N. J. Edmunds, S. Moncada, and J. M. Marshall, Does nitric oxide allow endothelial cells to sense hypoxia and mediate hypoxic vasodilation? In vivo and in vitro studies, J. Physiol. 546, 521–577 (2003).PubMedCrossRefGoogle Scholar

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© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Silvia Bertuglia
  • Andrea Giusti

There are no affiliations available

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