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Oxidative stress-induced iron signaling is responsible for peroxide-dependent oxidation of dichlorodihydrofluorescein in endothelial cells: role of transferrin receptor-dependent iron uptake in apoptosis. Circ Res 2003 Jan 10;92(1):56-63

Date

01/11/2003

Pubmed ID

12522121

DOI

10.1161/01.res.0000048195.15637.ac

Scopus ID

2-s2.0-0037428060 (requires institutional sign-in at Scopus site)   143 Citations

Abstract

Dichlorodihydrofluorescein (DCFH) is one of the most frequently used probes for detecting intracellular oxidative stress. In this study, we report that H2O2-dependent intracellular oxidation of DCFH to a green fluorescent product, 2',7'-dichlorofluorescein (DCF), required the uptake of extracellular iron transported through a transferrin receptor (TfR) in endothelial cells. H2O2-induced DCF fluorescence was inhibited by the monoclonal IgA-class anti-TfR antibody (42/6) that blocked TfR endocytosis and the iron uptake. H2O2-mediated inactivation of cytosolic aconitase was responsible for activation of iron regulatory protein-1 and increased expression of TfR, resulting in an increased iron uptake into endothelial cells. H2O2-mediated caspase-3 proteolytic activation was inhibited by anti-TfR antibody. Similar results were obtained in the presence of a lipid hydroperoxide. We conclude that hydroperoxide-induced DCFH oxidation and endothelial cell apoptosis required the uptake of extracellular iron by the TfR-dependent iron transport mechanism and that the peroxide-induced iron signaling, in general, has broader implications in oxidative vascular biology.

Author List

Tampo Y, Kotamraju S, Chitambar CR, Kalivendi SV, Keszler A, Joseph J, Kalyanaraman B

Authors

Balaraman Kalyanaraman PhD Professor in the Biophysics department at Medical College of Wisconsin
Agnes Keszler PhD Research Scientist I in the Biophysics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Aconitate Hydratase
Animals
Antibodies, Monoclonal
Apoptosis
Caspase 3
Caspases
Cattle
Cells, Cultured
Endocytosis
Endothelium, Vascular
Enzyme Activation
Fluoresceins
Fluorescent Dyes
Glutathione
Hydrogen Peroxide
Iron
Iron Chelating Agents
Iron Regulatory Protein 1
Lipid Peroxides
Metalloporphyrins
Oxidants
Oxidation-Reduction
Oxidative Stress
Receptors, Transferrin
Signal Transduction