Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1994 Oct;94(4):1621–1628. doi: 10.1172/JCI117504

Reperfusion injury induces apoptosis in rabbit cardiomyocytes.

R A Gottlieb 1, K O Burleson 1, R A Kloner 1, B M Babior 1, R L Engler 1
PMCID: PMC295322  PMID: 7929838

Abstract

The most effective way to limit myocardial ischemic necrosis is reperfusion, but reperfusion itself may result in tissue injury, which has been difficult to separate from ischemic injury. This report identifies elements of apoptosis (programmed cell death) in myocytes as a response to reperfusion but not ischemia. The hallmark of apoptosis, nucleosomal ladders of DNA fragments (approximately 200 base pairs), was detected in ischemic/reperfused rabbit myocardial tissue but not in normal or ischemic-only rabbit hearts. Granulocytopenia did not prevent nucleosomal DNA cleavage. In situ nick end labeling demonstrated DNA fragmentation predominantly in myocytes. The pattern of nuclear chromatin condensation was distinctly different in reperfused than in persistently ischemic tissue by transmission electron microscopy. Apoptosis may be a specific feature of reperfusion injury in cardiac myocytes, leading to late cell death.

Full text

PDF
1627

Images in this article

1625Image
on p.1625
1625Image
on p.1625
1625Image
on p.1625
1626Image
on p.1626
1626Image
on p.1626
1626Image
on p.1626
1623Image
on p.1623
1627Image
on p.1627

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Braunwald E., Kloner R. A. Myocardial reperfusion: a double-edged sword? J Clin Invest. 1985 Nov;76(5):1713–1719. doi: 10.1172/JCI112160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bursch W., Kleine L., Tenniswood M. The biochemistry of cell death by apoptosis. Biochem Cell Biol. 1990 Sep;68(9):1071–1074. doi: 10.1139/o90-160. [DOI] [PubMed] [Google Scholar]
  3. Cotter T. G., Lennon S. V., Glynn J. G., Martin S. J. Cell death via apoptosis and its relationship to growth, development and differentiation of both tumour and normal cells. Anticancer Res. 1990 Sep-Oct;10(5A):1153–1159. [PubMed] [Google Scholar]
  4. Dipasquale B., Youle R. J. Programmed cell death in heterokaryons. A study of the transfer of apoptosis between nuclei. Am J Pathol. 1992 Dec;141(6):1471–1479. [PMC free article] [PubMed] [Google Scholar]
  5. Engler R. L., Schmid-Schönbein G. W., Pavelec R. S. Leukocyte capillary plugging in myocardial ischemia and reperfusion in the dog. Am J Pathol. 1983 Apr;111(1):98–111. [PMC free article] [PubMed] [Google Scholar]
  6. Engler R., Gilpin E. Can superoxide dismutase alter myocardial infarct size? Circulation. 1989 May;79(5):1137–1142. doi: 10.1161/01.cir.79.5.1137. [DOI] [PubMed] [Google Scholar]
  7. Entman M. L., Michael L., Rossen R. D., Dreyer W. J., Anderson D. C., Taylor A. A., Smith C. W. Inflammation in the course of early myocardial ischemia. FASEB J. 1991 Aug;5(11):2529–2537. doi: 10.1096/fasebj.5.11.1868978. [DOI] [PubMed] [Google Scholar]
  8. Farb A., Kolodgie F. D., Jenkins M., Virmani R. Myocardial infarct extension during reperfusion after coronary artery occlusion: pathologic evidence. J Am Coll Cardiol. 1993 Apr;21(5):1245–1253. doi: 10.1016/0735-1097(93)90253-w. [DOI] [PubMed] [Google Scholar]
  9. Gaido M. L., Cidlowski J. A. Identification, purification, and characterization of a calcium-dependent endonuclease (NUC18) from apoptotic rat thymocytes. NUC18 is not histone H2B. J Biol Chem. 1991 Oct 5;266(28):18580–18585. [PubMed] [Google Scholar]
  10. Garlick P. B., Davies M. J., Hearse D. J., Slater T. F. Direct detection of free radicals in the reperfused rat heart using electron spin resonance spectroscopy. Circ Res. 1987 Nov;61(5):757–760. doi: 10.1161/01.res.61.5.757. [DOI] [PubMed] [Google Scholar]
  11. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Giannakis C., Forbes I. J., Zalewski P. D. Ca2+/Mg(2+)-dependent nuclease: tissue distribution, relationship to inter-nucleosomal DNA fragmentation and inhibition by Zn2+. Biochem Biophys Res Commun. 1991 Dec 16;181(2):915–920. doi: 10.1016/0006-291x(91)91278-k. [DOI] [PubMed] [Google Scholar]
  13. Gobé G. C., Axelsen R. A., Searle J. W. Cellular events in experimental unilateral ischemic renal atrophy and in regeneration after contralateral nephrectomy. Lab Invest. 1990 Dec;63(6):770–779. [PubMed] [Google Scholar]
  14. Hockenbery D. M., Zutter M., Hickey W., Nahm M., Korsmeyer S. J. BCL2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6961–6965. doi: 10.1073/pnas.88.16.6961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kerr J. F., Wyllie A. H., Currie A. R. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972 Aug;26(4):239–257. doi: 10.1038/bjc.1972.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kloner R. A., Przyklenk K., Whittaker P. Deleterious effects of oxygen radicals in ischemia/reperfusion. Resolved and unresolved issues. Circulation. 1989 Nov;80(5):1115–1127. doi: 10.1161/01.cir.80.5.1115. [DOI] [PubMed] [Google Scholar]
  17. Kukielka G. L., Hawkins H. K., Michael L., Manning A. M., Youker K., Lane C., Entman M. L., Smith C. W., Anderson D. C. Regulation of intercellular adhesion molecule-1 (ICAM-1) in ischemic and reperfused canine myocardium. J Clin Invest. 1993 Sep;92(3):1504–1516. doi: 10.1172/JCI116729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lennon S. V., Martin S. J., Cotter T. G. Dose-dependent induction of apoptosis in human tumour cell lines by widely diverging stimuli. Cell Prolif. 1991 Mar;24(2):203–214. doi: 10.1111/j.1365-2184.1991.tb01150.x. [DOI] [PubMed] [Google Scholar]
  19. Litt M. R., Jeremy R. W., Weisman H. F., Winkelstein J. A., Becker L. C. Neutrophil depletion limited to reperfusion reduces myocardial infarct size after 90 minutes of ischemia. Evidence for neutrophil-mediated reperfusion injury. Circulation. 1989 Dec;80(6):1816–1827. doi: 10.1161/01.cir.80.6.1816. [DOI] [PubMed] [Google Scholar]
  20. Liu G. S., Thornton J., Van Winkle D. M., Stanley A. W., Olsson R. A., Downey J. M. Protection against infarction afforded by preconditioning is mediated by A1 adenosine receptors in rabbit heart. Circulation. 1991 Jul;84(1):350–356. doi: 10.1161/01.cir.84.1.350. [DOI] [PubMed] [Google Scholar]
  21. Mangan D. F., Wahl S. M. Differential regulation of human monocyte programmed cell death (apoptosis) by chemotactic factors and pro-inflammatory cytokines. J Immunol. 1991 Nov 15;147(10):3408–3412. [PubMed] [Google Scholar]
  22. McCord J. M. Oxygen-derived radicals: a link between reperfusion injury and inflammation. Fed Proc. 1987 May 15;46(7):2402–2406. [PubMed] [Google Scholar]
  23. Migliorati G., Pagliacci C., Moraca R., Crocicchio F., Nicoletti I., Riccardi C. Interleukins modulate glucocorticoid-induced thymocyte apoptosis. Int J Clin Lab Res. 1992;21(4):300–303. doi: 10.1007/BF02591666. [DOI] [PubMed] [Google Scholar]
  24. Miura T., Downey J. M., Ooiwa H., Ogawa S., Adachi T., Noto T., Shizukuda Y., Iimura O. Progression of myocardial infarction in a collateral flow deficient species. Jpn Heart J. 1989 Sep;30(5):695–708. doi: 10.1536/ihj.30.695. [DOI] [PubMed] [Google Scholar]
  25. Murgia M., Pizzo P., Sandoná D., Zanovello P., Rizzuto R., Di Virgilio F. Mitochondrial DNA is not fragmented during apoptosis. J Biol Chem. 1992 Jun 5;267(16):10939–10941. [PubMed] [Google Scholar]
  26. Nayler W. G., Panagiotopoulos S., Elz J. S., Daly M. J. Calcium-mediated damage during post-ischaemic reperfusion. J Mol Cell Cardiol. 1988 Mar;20 (Suppl 2):41–54. doi: 10.1016/0022-2828(88)90331-8. [DOI] [PubMed] [Google Scholar]
  27. Sandford N. L., Searle J. W., Kerr J. F. Successive waves of apoptosis in the rat prostate after repeated withdrawal of testosterone stimulation. Pathology. 1984 Oct;16(4):406–410. doi: 10.3109/00313028409084731. [DOI] [PubMed] [Google Scholar]
  28. Savill J. S., Wyllie A. H., Henson J. E., Walport M. J., Henson P. M., Haslett C. Macrophage phagocytosis of aging neutrophils in inflammation. Programmed cell death in the neutrophil leads to its recognition by macrophages. J Clin Invest. 1989 Mar;83(3):865–875. doi: 10.1172/JCI113970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Savill J., Dransfield I., Hogg N., Haslett C. Vitronectin receptor-mediated phagocytosis of cells undergoing apoptosis. Nature. 1990 Jan 11;343(6254):170–173. doi: 10.1038/343170a0. [DOI] [PubMed] [Google Scholar]
  30. Schumer M., Colombel M. C., Sawczuk I. S., Gobé G., Connor J., O'Toole K. M., Olsson C. A., Wise G. J., Buttyan R. Morphologic, biochemical, and molecular evidence of apoptosis during the reperfusion phase after brief periods of renal ischemia. Am J Pathol. 1992 Apr;140(4):831–838. [PMC free article] [PubMed] [Google Scholar]
  31. Searle J., Kerr J. F., Bishop C. J. Necrosis and apoptosis: distinct modes of cell death with fundamentally different significance. Pathol Annu. 1982;17(Pt 2):229–259. [PubMed] [Google Scholar]
  32. Shirato C., Miura T., Ooiwa H., Toyofuku T., Wilborn W. H., Downey J. M. Tetrazolium artifactually indicates superoxide dismutase-induced salvage in reperfused rabbit heart. J Mol Cell Cardiol. 1989 Nov;21(11):1187–1193. doi: 10.1016/0022-2828(89)90695-0. [DOI] [PubMed] [Google Scholar]
  33. Smith C. W., Entman M. L., Lane C. L., Beaudet A. L., Ty T. I., Youker K., Hawkins H. K., Anderson D. C. Adherence of neutrophils to canine cardiac myocytes in vitro is dependent on intercellular adhesion molecule-1. J Clin Invest. 1991 Oct;88(4):1216–1223. doi: 10.1172/JCI115424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Smith E. F., 3rd, Egan J. W., Bugelski P. J., Hillegass L. M., Hill D. E., Griswold D. E. Temporal relation between neutrophil accumulation and myocardial reperfusion injury. Am J Physiol. 1988 Nov;255(5 Pt 2):H1060–H1068. doi: 10.1152/ajpheart.1988.255.5.H1060. [DOI] [PubMed] [Google Scholar]
  35. Tepper C. G., Studzinski G. P. Teniposide induces nuclear but not mitochondrial DNA degradation. Cancer Res. 1992 Jun 15;52(12):3384–3390. [PubMed] [Google Scholar]
  36. Ueda N., Shah S. V. Endonuclease-induced DNA damage and cell death in oxidant injury to renal tubular epithelial cells. J Clin Invest. 1992 Dec;90(6):2593–2597. doi: 10.1172/JCI116154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wijsman J. H., Jonker R. R., Keijzer R., van de Velde C. J., Cornelisse C. J., van Dierendonck J. H. A new method to detect apoptosis in paraffin sections: in situ end-labeling of fragmented DNA. J Histochem Cytochem. 1993 Jan;41(1):7–12. doi: 10.1177/41.1.7678025. [DOI] [PubMed] [Google Scholar]
  38. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]
  39. Wyllie A. H., Kerr J. F., Currie A. R. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251–306. doi: 10.1016/s0074-7696(08)62312-8. [DOI] [PubMed] [Google Scholar]
  40. Wyllie A. H., Morris R. G., Smith A. L., Dunlop D. Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis. J Pathol. 1984 Jan;142(1):67–77. doi: 10.1002/path.1711420112. [DOI] [PubMed] [Google Scholar]
  41. Youker K., Smith C. W., Anderson D. C., Miller D., Michael L. H., Rossen R. D., Entman M. L. Neutrophil adherence to isolated adult cardiac myocytes. Induction by cardiac lymph collected during ischemia and reperfusion. J Clin Invest. 1992 Feb;89(2):602–609. doi: 10.1172/JCI115626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zakeri Z. F., Quaglino D., Latham T., Lockshin R. A. Delayed internucleosomal DNA fragmentation in programmed cell death. FASEB J. 1993 Mar;7(5):470–478. doi: 10.1096/fasebj.7.5.8462789. [DOI] [PubMed] [Google Scholar]
  43. Zweier J. L., Flaherty J. T., Weisfeldt M. L. Direct measurement of free radical generation following reperfusion of ischemic myocardium. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1404–1407. doi: 10.1073/pnas.84.5.1404. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES