Skip to main content
SpringerLink
Log in

Effect of iron overload in the isolated ischemic and reperfused rat heart

  • Clinical Studies
  • Published:
Cardiovascular Drugs and Therapy Aims and scope Submit manuscript

Summary

It has been suggested that iron might play a pivotal role in the development of reperfusion-induced cellular injury through the activation of oxygen free radical producing reactions. The present study examined the effects of myocardial iron overload on cardiac vulnerability to ischemia and reperfusion. Moreover, the effect of the iron chelator deferoxamine in reversing ischemia-reperfusion injury was studied. Animals were treated with iron dextran solution (i.m. injection, 25 mg every third day during a 5 week period). The control group received the same treatment without iron. Isolated rat hearts were perfused at constant flow (11 ml/min) and subjected to a 15 minute period of global normothermic ischemia followed by reperfusion for 15 minutes. The effects of iron overload were investigated using functional and biochemical parameters, as well as ultrastructural characteristics of the ischemic-reperfused myocardium compared with placebo values. The results suggest that (a) a significant iron overload was obtained in plasma and hepatic and cardiac tissues (×2.5, ×16, and ×8, respectively) after chronic intramuscular administration of iron dextran (25 mg); (b) during normoxia, iron overload was associated with a slight reduction in cardiac function and an increase in lactate dehydrogenase (LDH) release (×1.5); (c) upon reperfusion, functional recovery was similar whether the heart had been subjected to iron overload or not. However, in the control group left ventricular end-diastolic pressure remained higher than in preischemic conditions, an effect that was not observed in the iron-overloaded group. Moreover, LDH release was markedly increased in the iron-loaded group (×4.2); (d) iron overload was associated with a significant worsening of the structural alterations observed during reperfusion, particularly at the mitochondrial and sarcomere level; (e) after 15 minutes of reperfusion, the activity of the anti-free-radical enzyme, glutathione peroxidase (GPX), was significantly reduced in ironoverloaded hearts, whereas catalase activity was increased; (e) the overall modifications observed in the presence of iron overload were prevented by deferoxamine. In conclusion, this study underlines the possible role of cardiac iron in the development of injury associated with ischemia and reperfusion, and the possible importance of the use of an iron-chelating agent in antiischemic therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Gauduel Y, Duvelleroy MA. Role of oxygen radicals in cardiac injury due to reoxygenation.J Mol Cell Cardiol 1984;16:459–470.

    Google Scholar 

  2. Guarnieri C, Flamigni F, Caldarera CM. Role of oxygen in the cellular damage induced by reoxygenation of hypoxic heart.J Mol Cell Cardiol 1980;12:797–808.

    Google Scholar 

  3. Arroyo CM, Kramer JH, Dickens BF, Weglicki WB. Identification of free radicals in myocardial ischemia/reperfusion by spin-trapping with nitrone DMPO.Febs Lett 1987;221:101–104.

    Google Scholar 

  4. Zweier JL, Flaherty JT, Weisfeldt ML. Direct measurement of free radical generation following reperfusion of ischemic myocardium.Proc Natl Acad Sci USA 1987;84:1404–1407.

    Google Scholar 

  5. Koller PT, Bergmann SR. Reduction of lipid peroxidation in reperfused isolated rabbit hearts by diltiazem.Circ Res 65:838–846.

  6. Ambrosio G, Flaherty JT, Duilio C, et al. Oxygen radicals generated at reflow induce peroxidation of membrane lipids in reperfused hearts.J Clin Invest 1991;87:2056–2066.

    Google Scholar 

  7. Halliwell B. Superoxide, iron, vascular endothelium and reperfusion injury.Free Radical Res Commun 1989:5:315–318.

    Google Scholar 

  8. Farber NE, Vercellotti GM, Jacob HS, et al. Evidence for a role of iron-catalysed oxidants in functional and metabolic stunning in the canine heart.Circ Res 1988;63:351–360.

    Google Scholar 

  9. Halliwell B. Superoxide dependent formation of hydroxyl radicals in the presence of iron salts is a feasible source of hydroxyl radicals in vivo.Biochem J 1982;205:461–467.

    Google Scholar 

  10. Halliwell B, Gutteridge JM. Iron as biological pro-oxidant.ISI Atlas of Science Biochemistry, pp 48–52.

  11. Fontecave M, Pierre JL. Iron metabolism: The low-molecular-mass iron pool.Biol Metals 1991;4:133–135.

    Google Scholar 

  12. Koster JF, Slee RG. Ferritin: A physiological iron donor for microsomal lipid peroxidation.Febs Lett 1986;199:85–88.

    Google Scholar 

  13. Boucher F, Pucheu S, Coudray C, et al. Evidence of cytosolic iron release during post-ischaemic reperfusion of isolated rat hearts: Influence on spin-trapping experiments with DMPO.Febs Lett 1992;302:261–264.

    Google Scholar 

  14. Bernier M, Hearse DJ, Manning AS. Reperfusion-induced arrhythmias and oxygen-derived free radicals. Studies with anti-free radical interventions and a free-radical generating system in the isolated perfused rat heart.Circ Res 1986;58:331–340.

    Google Scholar 

  15. Karwatowska-Prokopczuk E, Czarnowska E, Beresewiez A. Iron availability and free radical induced injury in the isolated ischaemic-reperfused rat heart.Cardiovasc Res 1992;26:58–66.

    Google Scholar 

  16. Myers CL, Weiss SJ, Kirsh MM, et al. Effects of supplementing hypothermic crystalloid cardioplegic solution with catalase, superoxide dismutase, allopurinol, or deferoxamine on functional recovery of globally ischemic and reperfused isolated hearts.J Thor Cardiovasc Surg 1986;91:281–289.

    Google Scholar 

  17. Mergner GW, Weglicki WB, Kramer JH. Postischemic free radical production in the venous blood of the regionally ischemic swine heart: Effect of Deferoxamine.Circulation 1991;84:2070–2090.

    Google Scholar 

  18. Ambrosio G, Zweier JL, Jacobus WE, et al. Improvement of postischemic myocardial function and metabolism induced by administration of deferoxamine at the time of reflow: The role of iron in the pathogenesis of reperfusion injury.Circulation 1987;76:906–915.

    Google Scholar 

  19. Bolli R, Patel BS, Zhu WX, et al. The iron chelator desferrioxamine attenuates postischemic ventricular dysfunction.Am J Physiol 1987;253:1372–1380.

    Google Scholar 

  20. Olson AD, Hamlin WB. A new method for serum iron and total iron binding capacity by atomic absorption spectrophotometry.Clin Chem 1969;15:438.

    Google Scholar 

  21. Langendorff O. Untersuchungen am überlebenden Säugethierherzen.Pflügers Arch 1895;61:291.

    Google Scholar 

  22. Krebs HA, Henseleit K. Untersuchungen über die Harnstoffbildung im Tierköper.Hoppe Seyler's Z 1932:210:33–66.

    Google Scholar 

  23. Kim BK, Huebers H, Pippart MJ, Finc CA. Storage iron exchange in the rat as affected by deferoxamine.J Lab Clin Med 1985;105:440–448.

    Google Scholar 

  24. Wroblewski F, La Due JS. Lactic dehydrogenase activity in blood.Proc Soc Biol Med 1955;90:210–213.

    Google Scholar 

  25. Marklund SL. Spectrophotometric study of spontaneous disproportionation of superoxide anion radical and sensitive direct assay for superoxide dismutase.J Biol Chem 1976;251:7504–7507.

    Google Scholar 

  26. Beers RF, Sizer IW. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase.J Biol Chem 1952;195:133–135.

    Google Scholar 

  27. Flohe L, Günzler WA. Assays of glutathione peroxidase.Meth Enzymol 1984;105:114–121.

    Google Scholar 

  28. Feuvray D, de Leiris J. Ultrastructural modifications induced by reoxygenation in the anoxic isolated rat heart perfused without exogenous substrate.J Mol Cell Cardiol 1975;7:307–314.

    Google Scholar 

  29. Schaper J, Meiser E, Stammler G. Ultrastructural morphometric analysis of myocardium from dogs, rats, hamsters, mice and from human hearts.Circ Res 1985;56:377–391.

    Google Scholar 

  30. Lison L.Statistiques Appliquées à la Biologie Expérimentale. Gauthier-Villars, 1958:291–310.

  31. Van der Kraaij AM, Mostert LJ, van Eijk HG, Koster JH. Iron-load increases the susceptibility of rat hearts to oxygen reperfusion damage-protection by the antioxidant (+)- cyanidol-3 and deferoxamine.Circulation 1988;78:442–449.

    Google Scholar 

  32. Voogd A, Sluiter W, Van Eijk HG, Koster JF. Low molecular weight iron and the oxygen paradox in isolated rat hearts.J Clin Invest 1992;90:2050–2055.

    Google Scholar 

  33. Huebers WA, Brittenham GM, Siba EC, Finch CA. Absorption of carbonyl iron.J Lab Clin Med 1986;108:473–478.

    Google Scholar 

  34. Parc CH, Bacon BR, Brittenham GM, Tavill AS. Pathology of dietary carbonyl iron overload in rats.Lab Invest 1987;57:555–563.

    Google Scholar 

  35. Pucheu S, Coudray C, Favier A, de Leiris J. Iron-overload and peroxidative damage in rats.J Mol Cell Cardiol 1991;23(Suppl IV):149.

    Google Scholar 

  36. Powell SR, Hall D, Shih A. Copper loading of hearts increases postischemic reperfusion injury.Circ Res 1991;69:881–885.

    Google Scholar 

  37. Gale GR, Litchenberg WH, Smith AB, et al. Comparative iron mobilizing actions of deferoxamine, 1,2-dimethyl-3-hydroxypyrid-4-one and pyridoxal isonicatinoyl hydrazone in iron hydroxamate-loaded mice.Res Commun Chem Pathol Pharmacol 1991;73:299–313.

    Google Scholar 

  38. Keberle H. The biochemistry of desferrioxamine and its relation to iron metabolism.Ann NY Acad Sci 1964;119:758–768.

    Google Scholar 

  39. Aisen P, Litowsky I. Iron transport and storage proteins.Ann Rev Biochem 1980;49:357–393.

    Google Scholar 

  40. Biemond P, Swaak AJ, Van Eijk HG, Koster JF. Superoxide-dependent iron release from ferritin in inflammatory diseases.Free Radical Biol Med 1988;4:185–198.

    Google Scholar 

  41. Aisen. Physical biochemistry of transferrin--update 1984–1988. In: Loehr TM, eds.Iron Carriers and Iron Proteins. Weinheim, VCH Publishers, 1989:353–372.

    Google Scholar 

  42. Pre J. Metabolism normal et pathologique du fer.Feuillets de Biologie 1989;30:55–67.

    Google Scholar 

  43. Braunwald E, Kloner RA. The stunned myocardium: Prolonged, postischemic ventricular dysfunction.Circulation 1982;66:1146–1149.

    Google Scholar 

  44. Hershko C, Link G, Pinson A, et al. Iron loading and chelation as studied in a heart cell culture system.Haem 20:247–252.

  45. Hearse DJ. Cellular damage during myocardial ischaemia: Metabolic changes leading to enzyme leakage. In: Hearse DJ, de Leiris J (eds).Enzymes in Cardiology. Diagnosis and Research. Chichester: John Wiley and Sons 1979:445–460.

    Google Scholar 

  46. Boucher F, Pucheu S, Schatz C, et al. Cardioprotective effect of indapamide in experimental ischemia and reperfusion in rats.Gen Hypertens 218:261–263.

  47. Reimer KA, Jennings RB. Myocardial ischemia, hypoxia, and infarction. In: Fozzard HA, et al., eds.The Heart and Cardiovascular System. New York: Raven Press, 1986:1133–1201.

    Google Scholar 

  48. Van der Laarse A, Aljona JC, Zoet ACM, et al. A comparative study ischemia-anoxia induced impairment of myocytic structure and cardiac function in the isolated isovolumically-contracting perfused rat heart.Cardiovasc Res 12:768–778.

  49. Thomas C, Morchouse L, Aust S. Ferritin and superoxide-dependent lipid peroxidation.J Biol Chem 1985;260:3275.

    Google Scholar 

  50. Garlick PB, Davis MJ, Hearse DJ. Direct detection of free radicals in the reperfused rat heart using electron spin resonance spectroscopy.Circ Res 1987;61:757–760.

    Google Scholar 

  51. Shlafer M, Brosamer K, Forder JR. Cerium chloride as a histochemical marker of hydrogen peroxide in reperfused ischemic hearts.J Mol Cell Cardiol 1990;22:83–97.

    Google Scholar 

  52. Corden BJ. Hemosiderosis in rodents and the effect of acetohydroxamic acid on urinary iron excretion.Exp Hematol 1986;14:971–974.

    Google Scholar 

  53. Flynn DM, Hoffbrand AV, Politis D. Subcutaneous desferrioxamine. The effect of 3 years treatment on liver, iron serum ferritin, and comments on echocardiography. In: Cao A, Carcassi U, Rowley PT (eds).Thalassemia: Recent Advances in Detection and Treatment. New York: Alan R. Liss Publishing, 1985:347–355.

    Google Scholar 

  54. Marcus RE, Davies SC, Bantock HM. Desferrioxamine to improve cardiac function in iron overloaded patients with thalassaemia major.Lancet 1980;1:392.

    Google Scholar 

  55. Van Jaarsveld H, Kuyl JM, Alberts DW. The protective effect of deferoxamine on rat myocardial mitochondria is not prolonged after withdrawal of deferoxamine.Basic Res Cardiol 1992;87:47–53.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Physiologie Cellulaire Cardiaque, Université Joseph Fourier, BP53X, Grenoble Cedex, France

    S Pucheu, N Tresallet &  J de Leiris

  2. Laboratoire de Biochimie C, Centre Hospitalier Universitaire de Grenoble, Grenoble, France

    C Coudray & A Favier

Authors
  1. S Pucheu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C Coudray
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N Tresallet
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Favier
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J de Leiris
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pucheu, S., Coudray, C., Tresallet, N. et al. Effect of iron overload in the isolated ischemic and reperfused rat heart. Cardiovasc Drug Ther 7, 701–711 (1993). https://doi.org/10.1007/BF00877824

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00877824

Key Words

Search

Navigation