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Am J Physiol Heart Circ Physiol 282: H1907-H1914, 2002. First published January 31, 2002; doi:10.1152/ajpheart.00393.2001
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Vol. 282, Issue 5, H1907-H1914, May 2002

Burn plasma mediates cardiac myocyte apoptosis via endotoxin

Deborah L. Carlson1, Ellis Lightfoot Jr.1, Debora D. Bryant1, Sandra B. Haudek1, David Maass2, Jureta Horton2, and Brett P. Giroir1

1 Department of Pediatrics and 2 Department of Surgery The University of Texas Southwestern Medical Center, Dallas, Texas 75390

Thermal trauma is associated with cardiac myocyte apoptosis in vivo. To determine whether cardiac myocyte apoptosis could be secondary to burn-induced cytokines or inflammatory mediators, we investigated the effects of tumor necrosis factor-alpha (TNF-alpha ) and burn plasma on a murine cardiac myocyte cell line and primary culture myocytes. HL-1 cells were exposed to plasma isolated from burned or sham rats. Burn, but not sham plasma, induced significant increases in caspase-3 activity and DNA fragmentation. Similar results were obtained in primary culture rat myocytes. A dose-dependent increase in caspase-3 activity was observed when HL-1 cells were incubated with increasing concentrations of TNF-alpha . Even though TNF-alpha increased apoptosis, enzyme-linked immunosorbent assay detected no TNF-alpha in burn plasma. Burn plasma also failed to induce TNF-alpha mRNA, eliminating an autocrine mechanism of TNF-alpha secretion and binding. Also, treatment of burn plasma containing rhuTNFR:Fc failed to inhibit apoptosis. To examine the possibility that endotoxin within burn plasma might account for the apoptotic effect, burn plasma was preincubated with rBPI21. Caspase-3 activity was reduced to control levels. These data indicate that burn plasma induces apoptosis in cardiac myocytes via an endotoxin-dependent mechanism and suggest that systemic inhibition of endotoxin may provide a therapeutic approach for treatment of burn-associated cardiac dysfunction.

tumor necrosis factor-alpha ; thermal trauma; caspase-3; enzyme-linked immunosorbent assay


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