Basic Research in Cardiology

, Volume 86, Issue 3, pp 266–272 | Cite as

Is malondialdehyde a marker of the effect of oxygen free radicals in rat heart tissue?

  • György Ballagi-Pordány
  • J. Richter
  • M. Koltai
  • Z. Aranyi
  • G. Pogátsa
  • W. Schaper
Original Contributions


We tested the effect of exogenous purine derived free radicals and H2O2 VS ischemia and reperfusion on the thiobarbituric-acid (TBA)-reactive material and malondialdehyde (MDA) formation in isolated rat hearts using the thiobarbituric acid test and high performance lipid chromatography (HPLC). We could not detect increased thiobarbituric-acid-reactive material or MDA- production during 6 MM H2O2 infusion, during free radical generation by purine-derived free radicals, or using ischemia and reperfusion. Increased thiobarbituric-acid-reactive material and MDA tissue levels were detected only during infusion of 12 mM H2O2 (p<0.001). We conclude that the generally used thiobarbituric acid assay for MDA is susceptible to artifacts and unsuited as an indirect measure for low-to-medium-levels of oxygen free radicals. Using HPLC assay, which accurately measures MDA, no evidence was found that MDA is a primary and direct lipid peroxidation product of exogenous or endogenous reactive oxygen species.

Key words

Malondialdehyde thiobarbituric-acid-reactive material reactiveoxygen species isolatedrat heart reperfusion 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Brasch H, Schoenberg MH, Younes M (1989) No evidence for an increased lipid peroxidation during reoxygenation in Langendorff hearts and isolated atria of rats. J Mol Cell Cardiol 21:697–707Google Scholar
  2. 2.
    Buege JA, Aust SD (1978) Microsomal lipid peroxidation. Methods Enzymol 52:302–310Google Scholar
  3. 3.
    Bull AW, Marnett LJ (1985) Determination of malondialdehyde by ion-pairing high-performance liquide chromatography. Analyt Biochem 149:284–290Google Scholar
  4. 4.
    Burton KP, McCord JM, Ghai H (1984) Myocardial alterations due to free radical generation. Am J Physiol 246:H776-H783Google Scholar
  5. 5.
    Das DK, Engelman RM, Rousou JA, Breyer RH, Otani H, Lemeshow S (1986) Pathophysiology of superoxide radical as potential mediator of reperfusion injury in pig heart. Basic Res Cardiol 81:155–166Google Scholar
  6. 6.
    Del Maestro RF (1980) An approach to free radicals in medicine and biology. Acta Physiol Scand 492:153–168Google Scholar
  7. 7.
    Esterbauer H, Lang J, Zadravec S, Slater F (1984) Detection of malondialdehyde by high-performance liquid chromatography. In: Packer L (ed) Methods in Enzymology, Oxygen Radicals in Biological Systems, pp 105–319Google Scholar
  8. 8.
    Forman MB, Puett DW, Virmani R (1989) Endothelial and myocardial injury during ischemia and reperfusion: Pathogenesis and therapeutic implications. JACC 13:450–459Google Scholar
  9. 9.
    Gauduel Y, Duvelleroy MA (1984) Role of oxygen radicals in cardiac injury due to reoxygenation. J Mol Cell Cardiol 16:459–470Google Scholar
  10. 10.
    Guarnieri G, Flamigni F, Caldarera CM (1980) Role of oxygen in the cellular damage induced by re-oxygenation of hypoxic heart. J Mol Cell Cardiol 12:797–808Google Scholar
  11. 11.
    Haberland A, Schimke I, Papies B (1989) Does the measurement of xanthine oxidase/xanthine dehydrogenase ratio reflect the actual tissue content of these enzymes? Biomed Biochim Acta 48:114–117Google Scholar
  12. 12.
    Kaneko M, Beamish RE, Dhalla NS (1989) Depression of heart sarcolemmal Ca2+-pump activity by oxygen free radicals. Am J Physiol 256:H368-H374Google Scholar
  13. 13.
    Kuppusamy P, Zweier JL (1989) Characterization of free radical generation by xanthine oxidase/ evidence for hydroxyl radical generation. J Biol Chem 264:9880–9884Google Scholar
  14. 14.
    Lang J, Heckenast P, Esterbauer B (1984) Detection of malondialdehyde by HPLC. In: Walter de Gruyter (ed) Oxygen Radicals in Chemistry and Biology, Berlin New York, pp 351–354Google Scholar
  15. 15.
    Liedtke AJ, Mahar CO, Ytrehus K, Mjos OD (1984) Estimates of free radical production in rat and swine hearts: method and application of measuring malondialdehyde levels in fresh and frozen myocardium. Basic Res Cardiol 79:513–518Google Scholar
  16. 16.
    Marklund SL (1988) Role of toxic effects of oxygen in reperfusion damage. J Mol Cell Cardiol 20 (Suppl 2):23–30Google Scholar
  17. 17.
    McCord JM, Fridovich I (1969) Superoxide dismutase: An enzymic function for erythrocuprein. J Biol Chem 244:6049–6055Google Scholar
  18. 18.
    Myung-Suk K, Akera T (1987) O2 free radicals: cause of ischemia-reperfusion injury to cardiac NaK-ATPase. Am J Physiol 252:H252-H257Google Scholar
  19. 19.
    Neely JR, Rovetto MJ (1975) Techniques for perfusing isolated rat hearts. Methods in Enzymology 39:43–60Google Scholar
  20. 20.
    Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358Google Scholar
  21. 21.
    Reimer KA, Murry CE, Richard VJ (1989) The role of neutrophils and free radicals in the ischemiereperfused heart: Why the confusion and controversy? J Mol Cell Cardiol 21:1225–1239Google Scholar
  22. 22.
    Saugstad OD, Sanderud J (1989) Circulatory effects of oxygen radicals. Biomed Biochim Acta 48:20–24Google Scholar
  23. 23.
    Vander Heide RS, Sobotka PA, Ganote CE (1987) Effects of the free radical scavenger DMTU and mannitol on the oxygen paradox in perfused rat hearts. J Mol Cell Cardiol 19:615–625Google Scholar
  24. 24.
    Wiswedel I, Ulbricht O, Augustin W (1989) Studies of lipid peroxidation in isolated rat heart mitochondria. Biomed Biochim Acta 48:73–76Google Scholar
  25. 25.
    Wollenberger A, Ristau O, Schoffa G (1960) Eine einfache Technik der extrem schnellen Abkühlung größerer Gewebestücke. Pflügers Arch 270:399–412Google Scholar
  26. 26.
    Zweier JL, Duke SS, Kuppusamy P, Sylvester JT, Gabrielson EW (1989) Electron paramagnetic resonance evidence that cellular oxygen toxicity is caused by the generation of superoxide and hydroxyl free radicals. FEBS Letters, 252:12–16Google Scholar

Copyright information

© Steinkopff Verlag 1991

Authors and Affiliations

  • György Ballagi-Pordány
    • 1
  • J. Richter
    • 2
  • M. Koltai
    • 1
  • Z. Aranyi
    • 1
  • G. Pogátsa
    • 1
  • W. Schaper
    • 2
  1. 1.National Institute of CardiologyResearch DepartmentBudapestHungary
  2. 2.Max Planck InstituteDepartment of Experimental CardiologyBad NauheimGermany

Personalised recommendations