Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia

V. Lakshminarayanan, M. Lewallen, Nikolaos G. Frangogiannis, A. J. Evans, K. E. Wedin, L. H. Michael, M. L. Entman

Research output: Contribution to journalArticle

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Abstract

Chemokine expression is associated with reperfusion of infarcted myocardium in the setting of tissue necrosis, intense inflammation, and inflammatory cytokine release. The specific synthesis of monocyte chemotactic protein (MCP)-1 mRNA by cardiac venules in reperfused infarcts corresponded to the region where leukocytes normally localize. MCP-1 could be induced by exogenous tumor necrosis factor (TNF)-α or by postischemic cardiac lymph containing TNF-α. However, the release of TNF-α during early reperfusion did not explain the venular localization of MCP-1 induction. To better understand the factors mediating MCP-1 induction, we examined the role of ischemia/reperfusion in a model of brief coronary occlusion in which no necrosis or inflammatory response is seen. Adult mongrel dogs were subjected to 15 minutes of coronary occlusion and 5 hours of reperfusion. Ribonuclease protection assay revealed up-regulation of MCP-1 mRNA only in ischemic segments of reperfused canine myocardium. Pretreatment with the reactive oxygen scavenger N-(2-mercaptopropionyl)-glycine completely inhibited MCP-1 induction. In situ hybridization localized MCP-1 message to small venular endothelium in ischemic areas without myocyte necrosis. Gel shift analysis of nuclear extracts from the ischemic area showed enhanced DNA binding of the transcription factors AP-1 and nuclear factor (NF)-κB, crucial for MCP-1 expression, in ischemic myocardial regions. Immunohistochemical staining demonstrated reperfusion-dependent dent nuclear translocation of c-Jun and NF-κB (p65) in small venular endothelium, only in the ischemic regions of the myocardium, that was inhibited by N-(2-mercaptopropionyl)-glycine. In vitro, treatment of cultured canine jugular vein endothelial cells with the reactive oxygen intermediate H2O2 induced a concentration-dependent increase in MCP-1 mRNA levels, which was inhibited by the antioxidant N-acetyl-L-cysteine, a precursor of glutathione, but not pyrrolidine dithiocarbamate, an inhibitor of NF-κB and activator of AP-1. In contrast to our studies with infarction, incubation of canine jugular vein endothelial cells with postischemic cardiac lymph did not induce MCP-1 mRNA expression suggesting the absence of cytokine-mediated MCP-1 induction after a sublethal ischemic period. These results suggest that reactive oxygen intermediate generation, after a brief ischemic episode, is capable of inducing MCP-1 expression in venular endothelium through AP-1 and NF-κB. Short periods of ischemia/reperfusion, insufficient to produce a myocardial infarction, induce MCP-1 expression, potentially mediating angiogenesis in the ischemic noninfarcted heart.

Original languageEnglish (US)
Pages (from-to)1301-1311
Number of pages11
JournalAmerican Journal of Pathology
Volume159
Issue number4
StatePublished - 2001
Externally publishedYes

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Chemokine CCL2
Vascular Endothelium
Ischemia
Oxygen
Reperfusion
Transcription Factor AP-1
Endothelium
Canidae
Myocardium
Messenger RNA
Necrosis
Tumor Necrosis Factor-alpha
Coronary Occlusion
Jugular Veins
Lymph
Glycine
Endothelial Cells
Cytokines
Venules
Acetylcysteine

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Lakshminarayanan, V., Lewallen, M., Frangogiannis, N. G., Evans, A. J., Wedin, K. E., Michael, L. H., & Entman, M. L. (2001). Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia. American Journal of Pathology, 159(4), 1301-1311.

Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia. / Lakshminarayanan, V.; Lewallen, M.; Frangogiannis, Nikolaos G.; Evans, A. J.; Wedin, K. E.; Michael, L. H.; Entman, M. L.

In: American Journal of Pathology, Vol. 159, No. 4, 2001, p. 1301-1311.

Research output: Contribution to journalArticle

Lakshminarayanan, V, Lewallen, M, Frangogiannis, NG, Evans, AJ, Wedin, KE, Michael, LH & Entman, ML 2001, 'Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia', American Journal of Pathology, vol. 159, no. 4, pp. 1301-1311.
Lakshminarayanan, V. ; Lewallen, M. ; Frangogiannis, Nikolaos G. ; Evans, A. J. ; Wedin, K. E. ; Michael, L. H. ; Entman, M. L. / Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia. In: American Journal of Pathology. 2001 ; Vol. 159, No. 4. pp. 1301-1311.
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