Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1995 Feb;146(2):419–428.

Cytokine mRNA expression in postischemic/reperfused myocardium.

A Herskowitz 1, S Choi 1, A A Ansari 1, S Wesselingh 1
PMCID: PMC1869839  PMID: 7856752

Abstract

While the role of cytokines in mediating injury during hind limb skeletal muscle ischemia followed by reperfusion has recently been described, the role of cytokines in myocardial infarction and ischemia/reperfusion have remained relatively unexplored. We hypothesize that cytokines play an important role in the regulation of postischemic myocardial inflammation. This study reports the temporal sequence of proinflammatory cytokine gene expression in postischemic/reperfused myocardium and localizes interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha)-protein by immunostaining. Rats were subjected to either permanent left anterior descending (LAD) occlusion or to 35 minutes of LAD occlusion followed by reperfusion and sacrificed up to 7 days later. Rat-specific oligonucleotide probes were used to semiquantitatively assess the relative expression of mRNA for TNF-alpha, IL-1 beta, IL-2, IL-6, interferon-gamma (IFN-gamma), and transforming growth factor-beta 1 (TGF-beta 1) utilizing the reverse transcriptase-polymerase chain reaction amplification technique. Increased cardiac mRNA levels for all cytokines except IL-6 and IFN-gamma were measurable within 15 to 30 minutes of LAD occlusion and increased levels were generally sustained for 3 hours. During early reperfusion, mRNA levels for IL-6 and TGF-beta 1 were significantly reduced compared with permanent LAD occlusion. In both groups, cytokine mRNA levels all returned to baseline levels at 24 hours, while IL-1 beta, TNF-alpha, and TGF-beta 1 mRNA levels again rose significantly at 7 days only in animals with permanent LAD occlusion. Immunostaining for IL-1 beta and TNF-alpha protein revealed two patterns of reactivity: 1) microvascular staining for both IL-1 beta and TNF-alpha protein only in postischemic reperfused myocardium in early post-reperfusion time points; and 2) staining of infiltrating macrophages in healing infarct zones which was most prominent at 7 days after permanent LAD occlusion. These results provide evidence for local expression of cytokine mRNA in postischemic myocardium and suggest that regulation of local cytokine release is altered during the postischemic period.

Full text

PDF
422

Images in this article

422Figure 1
on p.422
424Figure 3
on p.424

Selected References

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

  1. Ansari A. A., Sundstrom J. B., Runnels H., Jensen P., Kanter K., Mayne A., Herskowitz A. The absence of constitutive and induced expression of critical cell-adhesion molecules on human cardiac myocytes. Its role in transplant rejection. Transplantation. 1994 Mar 27;57(6):942–949. doi: 10.1097/00007890-199403270-00029. [DOI] [PubMed] [Google Scholar]
  2. Brown J. M., Anderson B. O., Repine J. E., Shanley P. F., White C. W., Grosso M. A., Banerjee A., Bensard D. D., Harken A. H. Neutrophils contribute to TNF induced myocardial tolerance to ischaemia. J Mol Cell Cardiol. 1992 May;24(5):485–495. doi: 10.1016/0022-2828(92)91838-v. [DOI] [PubMed] [Google Scholar]
  3. Brown J. M., White C. W., Terada L. S., Grosso M. A., Shanley P. F., Mulvin D. W., Banerjee A., Whitman G. J., Harken A. H., Repine J. E. Interleukin 1 pretreatment decreases ischemia/reperfusion injury. Proc Natl Acad Sci U S A. 1990 Jul;87(13):5026–5030. doi: 10.1073/pnas.87.13.5026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chung M. K., Gulick T. S., Rotondo R. E., Schreiner G. F., Lange L. G. Mechanism of cytokine inhibition of beta-adrenergic agonist stimulation of cyclic AMP in rat cardiac myocytes. Impairment of signal transduction. Circ Res. 1990 Sep;67(3):753–763. doi: 10.1161/01.res.67.3.753. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Finkel M. S., Hoffman R. A., Shen L., Oddis C. V., Simmons R. L., Hattler B. G. Interleukin-6 (IL-6) as a mediator of stunned myocardium. Am J Cardiol. 1993 May 15;71(13):1231–1232. doi: 10.1016/0002-9149(93)90654-u. [DOI] [PubMed] [Google Scholar]
  7. Finkel M. S., Oddis C. V., Jacob T. D., Watkins S. C., Hattler B. G., Simmons R. L. Negative inotropic effects of cytokines on the heart mediated by nitric oxide. Science. 1992 Jul 17;257(5068):387–389. doi: 10.1126/science.1631560. [DOI] [PubMed] [Google Scholar]
  8. Glass J. D., Wesselingh S. L., Selnes O. A., McArthur J. C. Clinical-neuropathologic correlation in HIV-associated dementia. Neurology. 1993 Nov;43(11):2230–2237. doi: 10.1212/wnl.43.11.2230. [DOI] [PubMed] [Google Scholar]
  9. Gulick T., Chung M. K., Pieper S. J., Lange L. G., Schreiner G. F. Interleukin 1 and tumor necrosis factor inhibit cardiac myocyte beta-adrenergic responsiveness. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6753–6757. doi: 10.1073/pnas.86.17.6753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirsch A. T., Talsness C. E., Schunkert H., Paul M., Dzau V. J. Tissue-specific activation of cardiac angiotensin converting enzyme in experimental heart failure. Circ Res. 1991 Aug;69(2):475–482. doi: 10.1161/01.res.69.2.475. [DOI] [PubMed] [Google Scholar]
  11. Ikeda U., Ohkawa F., Seino Y., Yamamoto K., Hidaka Y., Kasahara T., Kawai T., Shimada K. Serum interleukin 6 levels become elevated in acute myocardial infarction. J Mol Cell Cardiol. 1992 Jun;24(6):579–584. doi: 10.1016/0022-2828(92)91042-4. [DOI] [PubMed] [Google Scholar]
  12. Kloner R. A. Does reperfusion injury exist in humans? J Am Coll Cardiol. 1993 Feb;21(2):537–545. doi: 10.1016/0735-1097(93)90700-b. [DOI] [PubMed] [Google Scholar]
  13. Lane J. R., Neumann D. A., Lafond-Walker A., Herskowitz A., Rose N. R. Role of IL-1 and tumor necrosis factor in coxsackie virus-induced autoimmune myocarditis. J Immunol. 1993 Aug 1;151(3):1682–1690. [PubMed] [Google Scholar]
  14. Lefer A. M., Tsao P., Aoki N., Palladino M. A., Jr Mediation of cardioprotection by transforming growth factor-beta. Science. 1990 Jul 6;249(4964):61–64. doi: 10.1126/science.2164258. [DOI] [PubMed] [Google Scholar]
  15. Löw-Friedrich I., Weisensee D., Mitrou P., Schoeppe W. Cytokines induce stress protein formation in cultured cardiac myocytes. Basic Res Cardiol. 1992 Jan-Feb;87(1):12–18. doi: 10.1007/BF00795385. [DOI] [PubMed] [Google Scholar]
  16. Mannisi J. A., Weisman H. F., Bush D. E., Dudeck P., Healy B. Steroid administration after myocardial infarction promotes early infarct expansion. A study in the rat. J Clin Invest. 1987 May;79(5):1431–1439. doi: 10.1172/JCI112971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oliveira I. C., Sciavolino P. J., Lee T. H., Vilcek J. Downregulation of interleukin 8 gene expression in human fibroblasts: unique mechanism of transcriptional inhibition by interferon. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9049–9053. doi: 10.1073/pnas.89.19.9049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pober J. S., Lapierre L. A., Stolpen A. H., Brock T. A., Springer T. A., Fiers W., Bevilacqua M. P., Mendrick D. L., Gimbrone M. A., Jr Activation of cultured human endothelial cells by recombinant lymphotoxin: comparison with tumor necrosis factor and interleukin 1 species. J Immunol. 1987 May 15;138(10):3319–3324. [PubMed] [Google Scholar]
  19. Schunkert H., Dzau V. J., Tang S. S., Hirsch A. T., Apstein C. S., Lorell B. H. Increased rat cardiac angiotensin converting enzyme activity and mRNA expression in pressure overload left ventricular hypertrophy. Effects on coronary resistance, contractility, and relaxation. J Clin Invest. 1990 Dec;86(6):1913–1920. doi: 10.1172/JCI114924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Seekamp A., Mulligan M. S., Till G. O., Smith C. W., Miyasaka M., Tamatani T., Todd R. F., 3rd, Ward P. A. Role of beta 2 integrins and ICAM-1 in lung injury following ischemia-reperfusion of rat hind limbs. Am J Pathol. 1993 Aug;143(2):464–472. [PMC free article] [PubMed] [Google Scholar]
  21. Seekamp A., Mulligan M. S., Till G. O., Ward P. A. Requirements for neutrophil products and L-arginine in ischemia-reperfusion injury. Am J Pathol. 1993 Apr;142(4):1217–1226. [PMC free article] [PubMed] [Google Scholar]
  22. Seekamp A., Till G. O., Mulligan M. S., Paulson J. C., Anderson D. C., Miyasaka M., Ward P. A. Role of selectins in local and remote tissue injury following ischemia and reperfusion. Am J Pathol. 1994 Mar;144(3):592–598. [PMC free article] [PubMed] [Google Scholar]
  23. Seekamp A., Warren J. S., Remick D. G., Till G. O., Ward P. A. Requirements for tumor necrosis factor-alpha and interleukin-1 in limb ischemia/reperfusion injury and associated lung injury. Am J Pathol. 1993 Aug;143(2):453–463. [PMC free article] [PubMed] [Google Scholar]
  24. Seko Y., Matsuda H., Kato K., Hashimoto Y., Yagita H., Okumura K., Yazaki Y. Expression of intercellular adhesion molecule-1 in murine hearts with acute myocarditis caused by coxsackievirus B3. J Clin Invest. 1993 Apr;91(4):1327–1336. doi: 10.1172/JCI116333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shreeniwas R., Koga S., Karakurum M., Pinsky D., Kaiser E., Brett J., Wolitzky B. A., Norton C., Plocinski J., Benjamin W. Hypoxia-mediated induction of endothelial cell interleukin-1 alpha. An autocrine mechanism promoting expression of leukocyte adhesion molecules on the vessel surface. J Clin Invest. 1992 Dec;90(6):2333–2339. doi: 10.1172/JCI116122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Strieter R. M., Kunkel S. L., Burdick M. D., Lincoln P. M., Walz A. The detection of a novel neutrophil-activating peptide (ENA-78) using a sensitive ELISA. Immunol Invest. 1992 Oct;21(6):589–596. doi: 10.3109/08820139209069393. [DOI] [PubMed] [Google Scholar]
  27. Weisman H. F., Bartow T., Leppo M. K., Marsh H. C., Jr, Carson G. R., Concino M. F., Boyle M. P., Roux K. H., Weisfeldt M. L., Fearon D. T. Soluble human complement receptor type 1: in vivo inhibitor of complement suppressing post-ischemic myocardial inflammation and necrosis. Science. 1990 Jul 13;249(4965):146–151. doi: 10.1126/science.2371562. [DOI] [PubMed] [Google Scholar]
  28. Weisman H. F., Bush D. E., Mannisi J. A., Weisfeldt M. L., Healy B. Cellular mechanisms of myocardial infarct expansion. Circulation. 1988 Jul;78(1):186–201. doi: 10.1161/01.cir.78.1.186. [DOI] [PubMed] [Google Scholar]
  29. Wesselingh S. L., Power C., Glass J. D., Tyor W. R., McArthur J. C., Farber J. M., Griffin J. W., Griffin D. E. Intracerebral cytokine messenger RNA expression in acquired immunodeficiency syndrome dementia. Ann Neurol. 1993 Jun;33(6):576–582. doi: 10.1002/ana.410330604. [DOI] [PubMed] [Google Scholar]
  30. Wu C. J., Lovett M., Wong-Lee J., Moeller F., Kitamura M., Goralski T. J., Billingham M. E., Starnes V. A., Clayberger C. Cytokine gene expression in rejecting cardiac allografts. Transplantation. 1992 Aug;54(2):326–332. doi: 10.1097/00007890-199208000-00024. [DOI] [PubMed] [Google Scholar]
  31. Yamada T., Matsumori A., Sasayama S. Therapeutic effect of anti-tumor necrosis factor-alpha antibody on the murine model of viral myocarditis induced by encephalomyocarditis virus. Circulation. 1994 Feb;89(2):846–851. doi: 10.1161/01.cir.89.2.846. [DOI] [PubMed] [Google Scholar]
  32. Youker K. A., Hawkins H. K., Kukielka G. L., Perrard J. L., Michael L. H., Ballantyne C. M., Smith C. W., Entman M. L. Molecular evidence for induction of intracellular adhesion molecule-1 in the viable border zone associated with ischemia-reperfusion injury of the dog heart. Circulation. 1994 Jun;89(6):2736–2746. doi: 10.1161/01.cir.89.6.2736. [DOI] [PubMed] [Google Scholar]
  33. 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]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES