Abstract
Specific reactive oxygen species (ROS) from different sources, might lead to different and even opposite, cellular effects. We studied the production of specific ROS resulting from the exposure of human umbilical veins endothelial cells (HUVEC) to H2O2 derived from the natural antioxidant epigallocathechin gallate (EGCG) or from the exposure to IL-1β using a fluorogenic probe and flow cytometry, and evaluated by western blot analysis and immunocytochemistry the associated expression of transcription factors sensitive to both inflammatory and oxidative stress, such as NF-κB and Nrf2, and some downstream activated genes such as cyclooxygenase-2 (COX-2) and hemeoxygenase 1 (HO-1). The results obtained showed that exogenously-generated H2O2 induce anti-inflammatory and antioxidant effects in HUVECs counteracting the pro-inflammatory and pro-oxidant effect of IL-1β related to the production of superoxide anions. The underlying mechanisms resulting from the extracellular production of H2O2, include (1) Nrf2 nuclear translocation and the enhanced expression of antioxidant enzymes such as HO-1, and (2) the previously unreported inhibition of NF-κB and COX-2 expression. Overall, these findings provide evidence that the production of specific reactive oxygen species finely tunes endothelial cell function and might be relevant for the reappraisal of the effects of exogenous antioxidants in the context of cardiovascular diseases.
Keywords:
Antioxidants; Endothelial cells; Interleukin-1β; NF-κB; Oxidative stress; Reactive oxygen species.
Copyright © 2015 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Active Transport, Cell Nucleus
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Anti-Inflammatory Agents, Non-Steroidal / adverse effects
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Anti-Inflammatory Agents, Non-Steroidal / metabolism*
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Antioxidants / adverse effects
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Antioxidants / metabolism*
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Catechin / adverse effects
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Catechin / analogs & derivatives
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Catechin / metabolism
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Cells, Cultured
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Cyclooxygenase 2 / chemistry
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Cyclooxygenase 2 / genetics
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Cyclooxygenase 2 / metabolism
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Dietary Supplements / adverse effects
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Endothelium, Vascular / cytology
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Endothelium, Vascular / immunology
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Endothelium, Vascular / metabolism*
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Gene Expression Regulation
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Heme Oxygenase-1 / antagonists & inhibitors
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Heme Oxygenase-1 / chemistry
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Heme Oxygenase-1 / genetics
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Heme Oxygenase-1 / metabolism
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Human Umbilical Vein Endothelial Cells / cytology
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Humans
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Hydrogen Peroxide / adverse effects
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Hydrogen Peroxide / metabolism
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Interleukin-1beta / antagonists & inhibitors*
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Interleukin-1beta / metabolism
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NF-E2-Related Factor 2 / agonists
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NF-E2-Related Factor 2 / antagonists & inhibitors
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NF-E2-Related Factor 2 / metabolism
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NF-kappa B p52 Subunit / agonists
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NF-kappa B p52 Subunit / antagonists & inhibitors
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NF-kappa B p52 Subunit / genetics
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NF-kappa B p52 Subunit / metabolism
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Oxidative Stress*
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Reactive Oxygen Species / adverse effects
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Reactive Oxygen Species / agonists*
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Reactive Oxygen Species / metabolism
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Superoxides / metabolism
Substances
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Anti-Inflammatory Agents, Non-Steroidal
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Antioxidants
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IL1B protein, human
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Interleukin-1beta
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NF-E2-Related Factor 2
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NF-kappa B p52 Subunit
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NFE2L2 protein, human
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NFKB2 protein, human
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Reactive Oxygen Species
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gallocatechin-7-gallate
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Superoxides
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Catechin
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Hydrogen Peroxide
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HMOX1 protein, human
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Heme Oxygenase-1
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Cyclooxygenase 2
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PTGS2 protein, human