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-induced oxidant generation and NF-
B activation
Department of Pharmacology, College of Medicine, The University of Illinois, Chicago, Illinois 60612
Because reactive oxygen species (ROS) can
function as second messengers and regulate the activation of the
transcription factor nuclear factor (NF)-
B, we investigated the
possible role of tumor necrosis factor-
(TNF-)-induced ROS
generation in endothelial cells in signaling E-selectin gene
transcription. We demonstrated that stimulation of human pulmonary
artery endothelial cells with TNF- (100 U/ml) resulted in ROS
production using the oxidant-sensitive dye 5 (and
6)-carboxy-2',7'-dichlorodihydrofluorescein diacetate bis(acetoxymethyl)ester. Pretreatment with
N-acetyl-L-cysteine (NAC)
or pyrrolidine dithiocarbamate (PDTC) for 0.5 h inhibited TNF--induced generation of ROS as well as activation of NF-B and
E-selectin mRNA and the cell surface protein expression. These findings
indicate that TNF- induces NF-B activation and the resultant
E-selectin gene expression by a pathway that involves formation of ROS
and that E-selectin expression can be inhibited by the antioxidant
action of NAC or PDTC. The results support the hypothesis that
generation of ROS in endothelial cells induced by proinflammatory
cytokines such as TNF- is a critical signal mediating E-selectin
expression.
endothelium; tumor necrosis factor-; reactive oxygen species; nuclear factor-B
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