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Menadione‐Induced Oxidative Stress in Bovine Heart Microvascular Endothelial Cells a
Author(s) -
KOSSENJANS WILHELM,
RYMASZEWSKI ZBIGNIEW,
BARANKIEWICZ JERZY,
BOBST ALBERT,
ASHRAF MUHAMMAD
Publication year - 1996
Publication title -
microcirculation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.793
H-Index - 83
eISSN - 1549-8719
pISSN - 1073-9688
DOI - 10.3109/10739689609146781
Subject(s) - menadione , oxidative stress , reactive oxygen species , superoxide , hydrogen peroxide , chemistry , superoxide dismutase , hydroxyl radical , biochemistry , endothelial stem cell , radical , microbiology and biotechnology , in vitro , biology , enzyme
ABSTRACT Objective: Oxidative stress from increased production of reactive oxygen species or decreased efficiency of inhibitory and scavenger systems may contribute to vascular injury. In this study, we developed an in vitro model of vascular injury by menadione‐induced oxidative stress in bovine heart microvascular endothelial cells. Methods: Oxidative stress was induced by exposure to menadione. Superoxide, hydrogen peroxide and hydroxyl radical formation was measured by superoxide dismutase‐inhibitable cytochrome c reduction, the dichlorofluorescin technique and the salicylate method, respectively. Electron paramagnetic‐spin resonance spectroscopy employing 5–5′‐dimethyl‐1‐pyrroline‐N‐oxide for superoxide trapping was used. Endothelial cell cytotoxicity was assessed by lactate dehydrogenase release. Results: Superoxide and hydroxyl radical were produced in a time‐ and concentration‐dependent fashion. Fluorescence in the presence of dichlorofluorescin confirmed hydrogen peroxide formation. Endothelial cell cytotoxicity became evident after 5 h of menadione treatment at concentrations of 100 μM. 3‐Aminobenzamide, a poly(ADP‐ribose)polymerase inhibitor, and dimethylthiourea, a hydrogen peroxide and hydroxyl radical scavenger, decreased menadione cytotoxicity, whereas deferoxamine, an inhibitor of hydroxyl radical formation, did not. Conclusions: The results suggest that menadione toxicity is mediated by poly(ADP‐ribose)polymerase activation via hydrogen peroxide formation and that menadione‐treated bovine heart microvessel endothelial cells provide a suitable in vitro model to study oxidative stress in endothelial cells.