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J. Biol. Chem., Vol. 276, Issue 18, 14924-14932, May 4, 2001
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From the Departments of Lipopolysaccharide (LPS) has been implicated as
the bacterial component responsible for much of the endothelial cell
injury/dysfunction associated with Gram-negative bacterial infections.
Protein synthesis inhibition is required to sensitize the endothelium
to lipopolysaccharide-induced apoptosis, suggesting that a constitutive
or inducible cytoprotective protein(s) is required for endothelial
survival. We have identified two known endothelial anti-apoptotic
proteins, c-FLIP and Mcl-1, the expression of which is decreased
markedly in the presence of cycloheximide. Decreased expression of both
proteins preceded apoptosis evoked by lipopolysaccharide + cycloheximide. Caspase inhibition protected against apoptosis, but not
the decreased expression of c-FLIP and Mcl-1, suggesting that they
exert protection upstream of caspase activation. Inhibition of the
degradation of these two proteins with the proteasome inhibitor,
lactacystin, prevented lipopolysaccharide + cycloheximide-induced
apoptosis. Similarly, lactacystin protected against endothelial
apoptosis induced by either tumor necrosis factor-
A Constitutive Cytoprotective Pathway Protects Endothelial Cells
from Lipopolysaccharide-induced Apoptosis*
§¶,,
,,
Medicine and
§ Surgery, University of Washington School of Medicine,
Seattle, Washington 98104 and the Department of Molecular
Pharmacology, Isis Pharmaceuticals, Carlsbad, California 92008
or
interleukin-1
in the presence of cycloheximide. That apoptosis could
be blocked in the absence of new protein synthesis by inhibition of the
proteasome degradative pathway implicates the requisite involvement of
a constitutively expressed protein(s) in the endothelial cytoprotective pathway. Finally, reduction of FLIP expression with antisense oligonucleotides sensitized endothelial cells to LPS killing, demonstrating a definitive role for FLIP in the protection of endothelial cells from LPS-induced apoptosis.
*
This work was supported by National Institutes of Health
Grants GM42686, GM07037, HL59969, and HL03174.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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