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© 1999 American Heart Association, Inc.
Original Contribution |
Tumor Necrosis Factor-
Contributes to Ischemia- and Reperfusion-Induced Endothelial Activation in Isolated Hearts
Presented in part at the 71st Scientific Sessions of the American Heart Association, Dallas, Tex, November 8–11, 1998.
From the Institute of Physiology (C.K., S.Z., B.F.B.) and Institute for Surgical Research (H.H.), Ludwig-Maximilians-University, Munich, Germany; Department of Physiology (A.G.), Heinrich Heine University Düsseldorf, Germany; Department of Pathology (D.A.W.), Stanford University, Palo Alto, Calif; Internal Medicine I (C.K., P.B.), Ludwig-Maximilians-University, Munich, Germany; and Cardiovascular Department (R.A.K.), Brigham and Women's Hospital, Boston, Mass.
Correspondence to Christian Kupatt, MD, Internal Medicine I, Ludwig-Maximilians-University, Marchioninistr. 15, 81377 Munich, Germany. E-mail c.kupatt{at}lrz-muenchen.de
Abstract—During myocardial
reperfusion, polymorphonuclear neutrophil (PMN) adhesion involving
the intercellular adhesion molecule-1 (ICAM-1) may lead to aggravation
and prolongation of reperfusion injury. We studied the role of early
tumor necrosis factor-
(TNF-) cleavage and nuclear factor-
B
(NF-B) activation on ICAM-1 expression and venular adhesion of PMN
in isolated hearts after ischemia (15 minutes) and reperfusion
(30 to 480 minutes). NF-B activation (electromobility shift assay)
was found after 30 minutes of reperfusion and up to 240 minutes. ICAM-1
mRNA, assessed by Northern blot, increased during the same interval.
Functional effect of newly synthesized adhesion molecules was found by
quantification (in situ fluorescence microscopy) of PMN, given
as bolus after ischemia, which became adherent to small
coronary venules (10 to 50 µm in diameter). After 480
minutes of reperfusion, ICAM-1–dependent PMN adhesion increased
2.5-fold compared with PMN adhesion obtained during acute reperfusion.
To study the influence of NF-B on PMN adhesion, we inhibited NF-B
activation by transfection of NF-B decoy
oligonucleotides into isolated hearts using
HJV-liposomes. Decoy NF-B but not control
oligonucleotides blocked ICAM-1 upregulation and
inhibited the subacute increase in PMN adhesion. Similar effects
were obtained using BB 1101 (10 µg), an inhibitor of
TNF- cleavage enzyme. These data suggest that ischemia and
reperfusion in isolated hearts cause liberation of TNF-, activation
of NF-B, and upregulation of ICAM-1, an adhesion molecule involved
in inflammatory response after ischemia and reperfusion.
Key Words: ischemia • reperfusion • cytokine • polymorphonuclear neutrophil
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