Molecular and Cellular Biochemistry

, Volume 232, Issue 1–2, pp 19–26 | Cite as

Early changes in myocardial antioxidant enzymes in rats treated with adriamycin

  • Timao Li
  • Igor Danelisen
  • Pawan K. Singal


Increased oxidative stress and antioxidant deficit have been suggested to play a major role in adriamycin-induced cardiomyopathy and congestive heart failure due to multiple treatments with adriamycin (doxorubicin). In this study, we investigated the acute effects of a single dose of adriamycin on myocardial antioxidant enzymes in rats. Adriamycin (2.5 mg/kg) was injected (i.p.) and myocardial antioxidant enzyme activities, mRNA abundance and protein levels at 1, 2, 4 and 24 h were examined. While manganese superoxide dismutase (MnSOD), glutathione peroxidase (GSHPx) and catalase (CAT) activities were not significantly changed, copper-zinc superoxide dismutase (CuZnSOD) activity was reduced at all time points and this change correlated with a decrease in its protein content. CuZnSOD mRNA was increased at 1 and 24 h. GSHPx mRNA and protein levels were transiently decreased by 20 and 25% respectively at 2 h. MnSOD mRNA was not significantly changed, but its protein levels were significantly decreased at 1 h. Lipid peroxidation was increased transiently at 1, 2 and 4 h. A transient depression in antioxidant enzyme as well as transient increase in oxidative stress with a single dose of adriamycin may precede more sustained changes seen with the repeated administration of the drug and contribute to the development of cardiomyopathy and heart failure.

antioxidants lipid peroxidation oxidative stress doxorubicin cardiomyopathy heart failure 


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  1. 1.
    Young RC, Ozols RF, Myers CE: The anthracycline antineoplastic drugs. N Engl J Med 305: 139–153, 1981Google Scholar
  2. 2.
    Lefrak EA, Pitha J, Rosenheim S, Gottlieb JA: A clinicopathologic analysis of adriamycin cardiotoxicity. Cancer 32: 302–314, 1973Google Scholar
  3. 3.
    Singal PK, Iliskovic N: Doxorubicin-induced cardiomyopathy. N Engl J Med 339: 900–905, 1998Google Scholar
  4. 4.
    Myers CE, Mcguire WP, Liss RH, Ifrim I, Grotzinger K, Young RC: Adriamycin: The role of lipid peroxidation in cardiac toxicity and tumor response. Science 197: 165–167, 1977Google Scholar
  5. 5.
    Singal PK, Deally CM, Weinberg LE: Subcellular effects of adriamycin in the heart: A concise review. J Mol Cell Cardiol 19: 817–828, 1987Google Scholar
  6. 6.
    Singal PK, Iliskovic N, Li T, Kumar D: Adriamycin cardiomyopathy: Pathophysiology and prevention. FASEB J 11: 931–936, 1997Google Scholar
  7. 7.
    Singal PK, Petkau A, Gerrard JM, Hrushovetz S, Foerster J: Free radicals in health and disease. Mol Cell Biochem 84: 121–122, 1988Google Scholar
  8. 8.
    Cross CE, Halliwell B, Borish ET, Pryor WA, Ames BN, Saul RL, McCord JM, Harman D: Oxygen radicals and human disease. Ann Intern Med 107: 526–545, 1987Google Scholar
  9. 9.
    Freeman BA, Crapo JD: Biology of disease: Free radicals and tissue injury. Lab Invest 47: 412–426, 1982Google Scholar
  10. 10.
    Iliskovic N, Hasinoff BB, Malisza KL, Li T, Danelisen I, Singal PK: Mechanisms of beneficial effects of probucol in adriamycin cardiomyopathy. Mol Cell Biochem 196: 43–49, 1999Google Scholar
  11. 11.
    Kaul N, Siveski-Iliskovic N, Hill M, Slezak J, Singal PK: Free radicals and the heart. J Pharmacol Toxicol Meth 30: 55–67, 1993Google Scholar
  12. 12.
    Yen HC, Oberley TD, Vichitbandha S, Ho YS, St Clair DK: The protective role of manganese superoxide dismutase against adriamycininduced acute cardiac toxicity in transgenic mice. J Clin Invest 98: 1253–1260, 1996Google Scholar
  13. 13.
    Doroshow JH, Locker GY, Myers CE: Enzymatic defenses of the mouse heart against reactive oxygen metabolites: Alterations produced by doxorubicin. J Clin Invest 65: 128–135, 1980Google Scholar
  14. 14.
    Revis NW, Marusic N: Glutathione peroxidase activity and selenium concentration in the hearts of doxorubicin-treated rabbits. J Mol Cell Cardiol 10: 945–951, 1978Google Scholar
  15. 15.
    Siveski-Iliskovic N, Hill M, Chow DA, Singal PK: Probucol protects against adriamycin cardiomyopathy without interfering with its antitumor effect. Circulation 91: 10–15, 1995Google Scholar
  16. 16.
    Li T, Singal PK: Adriamycin-induced early changes in myocardial antioxidant enzymes and their modulation by probucol. Circulation 102: 2105–2110, 2000Google Scholar
  17. 17.
    Siveski-Iliskovic N, Kaul N, Singal PK: Probucol promotes endogenous antioxidants and provides protection against adriamycin-induced cardiomyopathy in rats. Circulation 89: 2829–2835, 1994Google Scholar
  18. 18.
    Marklund SL: Pyrogallol autooxidation. In: R.A. Greenwald (ed). Handbook of Methods for Oxygen Radical Research. Boca Raton, FL, CRC Press, 1985, pp 243–247Google Scholar
  19. 19.
    Geller BL, Winge DR: A method for distinguishing Cu, Zn-and Mn-containing superoxide dismutases. Anal Biochem 128: 86–92, 1983Google Scholar
  20. 20.
    Paglia DE, Valentine WN: Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70: 158–169, 1967Google Scholar
  21. 21.
    Clairborne A: Catalase activity. In: R.A. Greenwald (ed). Handbook of Methods for Oxygen Radical Research. Boca Raton, FL, CRC Press, 1985, pp 283–284Google Scholar
  22. 22.
    Singal PK, Pierce GN: Adriamycin stimulates low-affinity Ca2+ binding and lipid peroxidation but depresses myocardial function. Am J Physiol 250: H419–H425, 1986Google Scholar
  23. 23.
    Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987Google Scholar
  24. 24.
    Hurt J, Hsu JL, Dougall WC, Visner GA, Burr IM, Nick HS: Multiple mRNA species generated by alternate polyadenylation from the rat manganese superoxide dismutase gene. Nucleic Acids Res 20: 2985–2990, 1992Google Scholar
  25. 25.
    Lowry OH, Rosenbrough NT, Farr AL, Randall AT: Protein measurements with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951Google Scholar
  26. 26.
    Li T, Danelisen I, Bello-Klein A, Singal PK: Effects of probucol on changes of antioxidant enzymes in adriamycin-induced cardiomyopathy in rats. Cardiovasc Res 46: 523–530, 2000Google Scholar
  27. 27.
    Doroshow JH: Effect of anthracycline antibiotics on oxygen radical formation in rat heart. Cancer Res 43: 460–472, 1983Google Scholar
  28. 28.
    Kalyanaraman B, Perez-Reyes E, Mason RP: Spin-trapping and direct electron spin resonance investigations of the redox metabolism of quinone anticancer drugs. Biochim Biophys Acta 630: 119–130, 1980Google Scholar
  29. 29.
    Singal PK, Kirshenbaum LA: A relative deficit in antioxidant reserve may contribute in cardiac failure. Can J Cardiol 6: 47–49, 1990Google Scholar
  30. 30.
    Yin X, Wu H, Chen Y, Kang YJ: Induction of antioxidants by adriamycin in mouse heart. Biochem Pharmacol 56: 87–93, 1998Google Scholar
  31. 31.
    Maitre B, Jornot L, Junod AF: Effects of inhibition of catalase and superoxide dismutase activity on antioxidant enzyme mRNA levels. Am J Physiol 265: L636–L643, 1993Google Scholar
  32. 32.
    Jornot L, Junod AF: Hyperoxia, unlike phorbol ester, induces glutathione peroxidase through a protein kinase C-independent mechanism. Biochem J 326: 117–123, 1997Google Scholar
  33. 33.
    Dieterich S, Bieligk U, Beulich K, Hasenfuss G, Prestle J: Gene expression of antioxidative enzymes in the human heart. Increased expression of catalase in the end-stage of failing heart. Circulation 101: 33–39, 2000Google Scholar
  34. 34.
    Iqbal J, Clerch LB, Hass MA, Frank L, Massaro D: Endotoxin increases lung Cu, Zn superoxide dismutase mRNA: O2 raises enzyme synthesis. Am J Physiol 257: L61–L64, 1989Google Scholar
  35. 35.
    Tsan MF, White JE, Treanor C, Shaffer JB: Molecular basis for tumor necrosis factor-induced increase in pulmonary superoxide dismutase activities. Am J Physiol 259: L506–L512, 1990Google Scholar
  36. 36.
    Clerch LB, Massaro D: Tolerance of rats to hyperoxia. Lung antioxidant enzyme gene expression. J Clin Invest 91: 499–508, 1993Google Scholar
  37. 37.
    Okuno H, Akahori A, Sato H, Xanthoudakis S, Curran T, Iba H: Escape from redox regulation enhances the transforming activity of Fos. Oncogene 8: 695–701, 1993Google Scholar
  38. 38.
    Schreck R, Albermann K, Baeuerle PA: Nuclear factor κB: An oxidative stress-responsive transcription factor of eukaryotic cells. Free Radic Res Commun 17: 221–237, 1992Google Scholar
  39. 39.
    Toledano MB, Leonard WJ: Modulation of transcription factor NF-κB binding activity by oxidation-reduction in vitro. Proc Natl Acad Sci USA 88: 4328–4332, 1991Google Scholar
  40. 40.
    Stevens JB, Autor AP: Induction of superoxide dismutase by oxygen in neonatal rat lung. J Biol Chem 252: 3509–3514, 1977Google Scholar
  41. 41.
    Shull S, Heintz NH, Periasamy M, Manohar M, Janssen YM, Marsh JP, Mossman BT: Differential regulation of antioxidant enzymes in response to oxidants. J Biol Chem 266: 24398–24403, 1991Google Scholar
  42. 42.
    Pinkus R, Weiner LM, Daniel V: Role of oxidants and antioxidants in the induction of AP-1, NF-κB, and glutathione S-transferase gene expression. J Biol Chem 271: 13422–13429, 1996Google Scholar
  43. 43.
    Wong GH, Elwell JH, Oberley LW, Goeddel DV: Manganous superoxide dismutase is essential for cellular resistance to cytotoxicity of tumor necrosis factor. Cell 58: 923–931, 1989Google Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Timao Li
  • Igor Danelisen
  • Pawan K. Singal

There are no affiliations available

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