(Circulation. 1999;100:541-546.)
© 1999 American Heart Association, Inc.
Basic Science Reports |
Protective Effects of Preconditioning in Cultured Rat Endothelial Cells
Effects on Neutrophil Adhesion and Expression of ICAM-1 After Anoxia and Reoxygenation
From IFRMP 23, INSERM E9920 (P.B., V.R., F.T., F.L., C.T.), VACOMED, the Department of Pharmacology, Rouen University Medical School, INSERM U78 (J.-P.L., M.D.), INSERM U413 (H.V.), Rouen, France.
Correspondence to Vincent Richard, Service de Pharmacologie, CHU de Rouen, 76031 Rouen Cedex, France. E-mail vincent.richard{at}chu-rouen.fr
Background—Preconditioning with brief periods of ischemia protects the coronary endothelium against acute and chronic reperfusion injury, but the mechanisms of this endothelial protection remain unknown. We hypothesized that preconditioning protects endothelial cells through a decreased production of endothelial adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1), leading to a lesser adhesion of neutrophils to the endothelium.
Methods and Results—Cultured rat aortic endothelial cells were subjected to 6-hour anoxia followed by various durations of reoxygenation. Preconditioning was induced by 1-hour anoxia and 1-hour reoxygenation. ICAM-1 gene expression was measured by polymerase chain reaction, and the percentage of cells expressing ICAM-1 was assessed by confocal laser fluorescence microscopy. Anoxia/reoxygenation increased expression of ICAM-1, with a peak occurring after 6 hours of reoxygenation for mRNA and 9 hours for protein. Preconditioning prevented the increase in ICAM-1. Similar reductions were observed with the free radical scavenger N-2 mercaptopropionyl glycine (MPG). The inhibitory effect of preconditioning on ICAM-1 expression was abolished by an inhibitor of protein kinase C, an inhibitor of nitric oxide synthesis, and by MPG but was not affected by an adenosine receptor antagonist. Finally, both preconditioning and MPG partially prevented the increased adhesion of human neutrophils to reoxygenated endothelial cells.
Conclusions—Preconditioning prevented reoxygenation-induced, free radical–mediated expression of ICAM-1 by a mechanism involving activation of protein kinase C and production of nitric oxide and free radicals, and this is associated with a lesser adhesion of neutrophils to endothelial cells. Such prevention of neutrophil adhesion may contribute to the protective effect of preconditioning against reperfusion-induced endothelial injury.
Key Words: endothelium • ischemia • cell adhesion molecules • free radicals
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