Skip to main content
NCBI home page
Mediators of Inflammation logoLink to Mediators of Inflammation
. 1993;2(1):85–92. doi: 10.1155/S0962935193000122

The role of the neutrophil and formed elements of the blood in an in vitro model of reperfusion injury

John A Barrett 2,✉, Claudia K Derian 1, Robert S Swillo 1, Richard F Woltmann 1, Mark H Perrone 1
PMCID: PMC2365378  PMID: 18475508

Abstract

Using The globally ischaemic isolated guinea-pig heart we conducted studies to assess the role of activated neutrophils (PMNs) and the role of the endothelium in reperfusion injury. Reperfusion injury was induced by a 20 min period of global ischaemia followed by a 30 min reperfusion with Krebs' buffer supplemented with f-Met–Leu–Phe (fMLP) and heparinized blood. Ischaemia alone or blood alone resulted in a complete recovery in contractile function measured by developed pressure, fMLP (500 μM) and blood, administered to normoxic hearts did not affect contractile function. The combination of 100 μM fMLP and blood beginning at reperfusion and continuing for 30 min decreased the recovery in contractile function (max. 33 ± 6% reovery) while buffer and 100 pM fMLP resulted in a complete recovery in function. In hearts infused with buffer and neutropenic blood incubated with 100 μM fMLP a complete recovery in function was observed. Isolated peritoneal neutrophils, 7–70 × 105 PMN/ min, incubated with 100 μM fMLP and Krebs' solution decreased contractile function in a concentration-related manner (max. 44 ± 11% recovery). Platelets, plasma or red blood cells alone incubated with fMLP did not decrease recovery in developed pressure. Platelets and PMN incubated with 100 μM fMLP did not, while red blood cells and PMN did, elicit a reduction in recovery in contractile function (34 ± 4% recovery). A 20 min period of global ischaemia destroys the functional integrity of the endothelium (response to Ach). Pre-treatment of the heart with sufficient H2O2 to functionally damage the endothelium, followed by infusion of Krebs' solution supplemented with blood and 100 μM fMLP also elicited a reduction in recovery of contractile function (42 ± 15% recovery). In summary, partially activated neutrophils play a major role in reperfusion injury and there exists a cooperativity between the RBC and PMN in this model.

Full Text

The Full Text of this article is available as a PDF (772.4 KB).

Selected References

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

  1. Arfors K. E., Lundberg C., Lindbom L., Lundberg K., Beatty P. G., Harlan J. M. A monoclonal antibody to the membrane glycoprotein complex CD18 inhibits polymorphonuclear leukocyte accumulation and plasma leakage in vivo. Blood. 1987 Jan;69(1):338–340. [PubMed] [Google Scholar]
  2. Bagchi D., Das D. K., Engelman R. M., Prasad M. R., Subramanian R. Polymorphonuclear leucocytes as potential source of free radicals in the ischaemic-reperfused myocardium. Eur Heart J. 1990 Sep;11(9):800–813. doi: 10.1093/oxfordjournals.eurheartj.a059800. [DOI] [PubMed] [Google Scholar]
  3. Bednar M., Smith B., Pinto A., Mullane K. M. Neutrophil depletion suppresses 111In-labeled platelet accumulation in infarcted myocardium. J Cardiovasc Pharmacol. 1985 Sep-Oct;7(5):906–912. doi: 10.1097/00005344-198509000-00014. [DOI] [PubMed] [Google Scholar]
  4. Bevilacqua M. P., Pober J. S., Mendrick D. L., Cotran R. S., Gimbrone M. A., Jr Identification of an inducible endothelial-leukocyte adhesion molecule. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9238–9242. doi: 10.1073/pnas.84.24.9238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bogman M. J., Cornelissen I. M., Berden J. H., De Jong J., Koene R. A. A comparative study of total body irradiation as a method of inducing granulocyte depletion in mice. J Immunol Methods. 1984 May 11;70(1):31–38. doi: 10.1016/0022-1759(84)90386-7. [DOI] [PubMed] [Google Scholar]
  6. Braunwald E. The aggressive treatment of acute myocardial infarction. Circulation. 1985 Jun;71(6):1087–1092. doi: 10.1161/01.cir.71.6.1087. [DOI] [PubMed] [Google Scholar]
  7. Carden D. L., Smith J. K., Korthuis R. J. Neutrophil-mediated microvascular dysfunction in postischemic canine skeletal muscle. Role of granulocyte adherence. Circ Res. 1990 May;66(5):1436–1444. doi: 10.1161/01.res.66.5.1436. [DOI] [PubMed] [Google Scholar]
  8. Chu A., Cobb F. R. Reperfusion alters the relation between blood flow and the remaining myocardial infarction. Circulation. 1989 Apr;79(4):884–889. doi: 10.1161/01.cir.79.4.884. [DOI] [PubMed] [Google Scholar]
  9. Coëffier E., Delautier D., Le Couedic J. P., Chignard M., Denizot Y., Benveniste J. Cooperation between platelets and neutrophils for paf-acether (platelet-activating factor) formation. J Leukoc Biol. 1990 Mar;47(3):234–243. doi: 10.1002/jlb.47.3.234. [DOI] [PubMed] [Google Scholar]
  10. Engler R. L. Free radical and granulocyte-mediated injury during myocardial ischemia and reperfusion. Am J Cardiol. 1989 Mar 7;63(10):19E–23E. doi: 10.1016/0002-9149(89)90225-7. [DOI] [PubMed] [Google Scholar]
  11. Engler R., Covell J. W. Granulocytes cause reperfusion ventricular dysfunction after 15-minute ischemia in the dog. Circ Res. 1987 Jul;61(1):20–28. doi: 10.1161/01.res.61.1.20. [DOI] [PubMed] [Google Scholar]
  12. Fitzpatrick F., Haeggström J., Granström E., Samuelsson B. Metabolism of leukotriene A4 by an enzyme in blood plasma: a possible leukotactic mechanism. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5425–5429. doi: 10.1073/pnas.80.17.5425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Freed M. S., Needleman P., Dunkel C. G., Saffitz J. E., Evers A. S. Role of invading leukocytes in enhanced atrial eicosanoid production following rabbit left ventricular myocardial infarction. J Clin Invest. 1989 Jan;83(1):205–212. doi: 10.1172/JCI113860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harlan J. M. Neutrophil-mediated vascular injury. Acta Med Scand Suppl. 1987;715:123–129. doi: 10.1111/j.0954-6820.1987.tb09912.x. [DOI] [PubMed] [Google Scholar]
  15. Henderson D. K., Hockey L. J., Vukalcic L. J., Edwards J. E., Jr Effect of immunosuppression on the development of experimental hematogenous Candida endophthalmitis. Infect Immun. 1980 Feb;27(2):628–631. doi: 10.1128/iai.27.2.628-631.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kuehl F. A., Jr, Dougherty H. W., Ham E. A. Interactions between prostaglandins and leukotrienes. Biochem Pharmacol. 1984 Jan 1;33(1):1–5. doi: 10.1016/0006-2952(84)90362-9. [DOI] [PubMed] [Google Scholar]
  17. Lefer A. M., Aoki N. Leukocyte-dependent and leukocyte-independent mechanisms of impairment of endothelium-mediated vasodilation. Blood Vessels. 1990;27(2-5):162–168. doi: 10.1159/000158807. [DOI] [PubMed] [Google Scholar]
  18. Lewis M. S., Whatley R. E., Cain P., McIntyre T. M., Prescott S. M., Zimmerman G. A. Hydrogen peroxide stimulates the synthesis of platelet-activating factor by endothelium and induces endothelial cell-dependent neutrophil adhesion. J Clin Invest. 1988 Dec;82(6):2045–2055. doi: 10.1172/JCI113825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lucchesi B. R., Werns S. W., Fantone J. C. The role of the neutrophil and free radicals in ischemic myocardial injury. J Mol Cell Cardiol. 1989 Dec;21(12):1241–1251. doi: 10.1016/0022-2828(89)90670-6. [DOI] [PubMed] [Google Scholar]
  20. McGee J. E., Fitzpatrick F. A. Erythrocyte-neutrophil interactions: formation of leukotriene B4 by transcellular biosynthesis. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1349–1353. doi: 10.1073/pnas.83.5.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mehta J. L., Nichols W. W., Donnelly W. H., Lawson D. L., Saldeen T. G. Impaired canine coronary vasodilator response to acetylcholine and bradykinin after occlusion-reperfusion. Circ Res. 1989 Jan;64(1):43–54. doi: 10.1161/01.res.64.1.43. [DOI] [PubMed] [Google Scholar]
  22. Mileski W. J., Winn R. K., Vedder N. B., Pohlman T. H., Harlan J. M., Rice C. L. Inhibition of CD18-dependent neutrophil adherence reduces organ injury after hemorrhagic shock in primates. Surgery. 1990 Aug;108(2):206–212. [PubMed] [Google Scholar]
  23. Mueller H. W., O'Flaherty J. T., Wykle R. L. Biosynthesis of platelet activating factor in rabbit polymorphonuclear neutrophils. J Biol Chem. 1983 May 25;258(10):6213–6218. [PubMed] [Google Scholar]
  24. Mullane K. M., Westlin W., Kraemer R. Activated neutrophils release mediators that may contribute to myocardial injury and dysfunction associated with ischemia and reperfusion. Ann N Y Acad Sci. 1988;524:103–121. doi: 10.1111/j.1749-6632.1988.tb38534.x. [DOI] [PubMed] [Google Scholar]
  25. Mullane K., Hatala M. A., Kraemer R., Sessa W., Westlin W. Myocardial salvage induced by REV-5901: an inhibitor and antagonist of the leukotrienes. J Cardiovasc Pharmacol. 1987 Oct;10(4):398–406. doi: 10.1097/00005344-198710000-00004. [DOI] [PubMed] [Google Scholar]
  26. Quillen J. E., Sellke F. W., Brooks L. A., Harrison D. G. Ischemia-reperfusion impairs endothelium-dependent relaxation of coronary microvessels but does not affect large arteries. Circulation. 1990 Aug;82(2):586–594. doi: 10.1161/01.cir.82.2.586. [DOI] [PubMed] [Google Scholar]
  27. Reimer K. A., Murry C. E., Richard V. J. The role of neutrophils and free radicals in the ischemic-reperfused heart: why the confusion and controversy? J Mol Cell Cardiol. 1989 Dec;21(12):1225–1239. doi: 10.1016/0022-2828(89)90669-x. [DOI] [PubMed] [Google Scholar]
  28. Romson J. L., Hook B. G., Kunkel S. L., Abrams G. D., Schork M. A., Lucchesi B. R. Reduction of the extent of ischemic myocardial injury by neutrophil depletion in the dog. Circulation. 1983 May;67(5):1016–1023. doi: 10.1161/01.cir.67.5.1016. [DOI] [PubMed] [Google Scholar]
  29. Ruggeri Z. M., De Marco L., Gatti L., Bader R., Montgomery R. R. Platelets have more than one binding site for von Willebrand factor. J Clin Invest. 1983 Jul;72(1):1–12. doi: 10.1172/JCI110946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sasaki K., Ueno A., Katori M., Kikawada R. Detection of leukotriene B4 in cardiac tissue and its role in infarct extension through leucocyte migration. Cardiovasc Res. 1988 Feb;22(2):142–148. doi: 10.1093/cvr/22.2.142. [DOI] [PubMed] [Google Scholar]
  31. Simpson P. J., Todd R. F., 3rd, Fantone J. C., Mickelson J. K., Griffin J. D., Lucchesi B. R. Reduction of experimental canine myocardial reperfusion injury by a monoclonal antibody (anti-Mo1, anti-CD11b) that inhibits leukocyte adhesion. J Clin Invest. 1988 Feb;81(2):624–629. doi: 10.1172/JCI113364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weyrich A. S., Ma X. L., Lefer A. M. The role of L-arginine in ameliorating reperfusion injury after myocardial ischemia in the cat. Circulation. 1992 Jul;86(1):279–288. doi: 10.1161/01.cir.86.1.279. [DOI] [PubMed] [Google Scholar]
  33. de Lorgeril M., Basmadjian A., Lavallée M., Clément R., Millette D., Rousseau G., Latour J. G. Influence of leukopenia on collateral flow, reperfusion flow, reflow ventricular fibrillation, and infarct size in dogs. Am Heart J. 1989 Mar;117(3):523–532. doi: 10.1016/0002-8703(89)90724-2. [DOI] [PubMed] [Google Scholar]

Articles from Mediators of Inflammation are provided here courtesy of Wiley

RESOURCES