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
01/11/2003Pubmed ID
12522121DOI
10.1161/01.res.0000048195.15637.acScopus ID
2-s2.0-0037428060 (requires institutional sign-in at Scopus site) 143 CitationsAbstract
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 BAuthors
Balaraman Kalyanaraman PhD Professor in the Biophysics department at Medical College of WisconsinAgnes 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 HydrataseAnimals
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