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Oxidative Stress in a Clonal Cell Line of Neuronal Origin: Effects of Antioxidant Enzyme Modulation
Author(s) -
Buckman Trent D.,
Sutphin Mary S.,
Mitrovic Branislava
Publication year - 1993
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1993.tb03489.x
Subject(s) - oxidative stress , antioxidant , enzyme , cell culture , modulation (music) , oxidative phosphorylation , enzyme assay , microbiology and biotechnology , cell , biology , chemistry , biochemistry , genetics , physics , acoustics
The effects of intracellularly generated H 2 O 2 on cell viability, morphology, and biochemical markers of injury have been investigated in a clonal cell line of neuronal origin (140‐3, mouse neuroblastoma X rat glioma) as a cell culture model for the role of oxidative stress in the longterm loss of neurons in the brain. The H 2 O 2 was generated from the redox cycling of menadione, or by the oxidation of serotonin catalyzed by monoamine oxidase, to simulate the effect of amine neurotransmitter turnover. Incubation with menadione at concentrations as low as 10 γ M for several hours resulted in significant losses of cell viability and altered morphology. Similar effects were evident in the presence of serotonin only after incubation overnight with concentrations > 1 m M . The cytotoxicity of either agent was potentiated by preincubation with specific inhibitors of two enzymes important to cellular antioxidant defenses, 3‐amino‐1,2,4‐trazole for catalase and 1,3‐bis(chloromethyl)‐1‐nitrosourea for glutathione reductase. Activity of another antioxidant enzyme of particular importance to antioxidant defenses in brain, the selenoprotein glutathione peroxidase, was stimulated fourfold by growth of cultures in the presence of sodium selenite as a source of active‐site Se for the enzyme. The only effect of the selenite on other functionally coupled antioxidant enzymes was a decrease in activity of superoxide dismutase at concentrations >200 n M . The selenite substantially protected cells against oxidative stress induced by combinations of menadione, 3‐amino‐1,2,4‐trazole, and 1,3‐bis(chloromethyl)‐1‐nitrosourea, but was only marginally effective with serotonin as a source of oxidative stress. The monoamine oxidase inhibitor pargyline increased cell survival in the presence of serotonin, demonstrating the role of this enzyme in its cytotoxicity. DNA damage (single strand breaks), but not lipid peroxidation, correlated with the cytotoxic effects of menadione.