Role of reactive oxygen species and glutathione in inorganic mercury-induced injury in human glioma cells.
The present study was undertaken to examine the role of reactive oxygen species (ROS) and glutathione (GSH) in glia cells using human glioma cell line A172 cells. HgCl2 caused the loss of cell viability in a dose-dependent manner. HgCl2-induced loss of cell viability was not affected by H2O2 scavengers catalase and pyruvate, a superoxide scavenger superoxide dismutase, a peroxynitrite scavenger uric acid, and an inhibitor of nitric oxide N(G)-nitro-arginine Methyl ester. HgCl2 did not cause changes in DCF fluorescence, an H2O2-sensitive fluorescent dye. The loss of cell viability was significantly prevented by the hydroxyl radical scavengers dimethylthiourea and thiourea, but it was not affected by antioxidants DPPD and Trlox. HgCl2-induced loss of cell viability was accompanied by a significant reduction in GSH content. The GSH depletion was almost completely prevented by thiols dithiothreitol and GSH, whereas the loss of viability was partially prevented by these agents. Incubation of cells with 0.2 mM buthionine sulfoximine for 24 hr, a selective inhibitor of gamma-glutamylcysteine synthetase, resulted in 56% reduction in GSH content without any change in cell viability. HgCl2 resulted in 34% reduction in GSH content, which was accompanied by 59% loss of cell viability. These results suggest that HgCl2-induced cell death is not associated with generation of H2O2 and ROS-induced lipid peroxidation. In addition, these data suggest that the depletion of endogenous GSH itself may not play a critical role in the HgCl2-induced cytotoxicity in human glioma cells.
Lee
Y W
YW
Department of Neurosurgery, College of Medicine, Pusan National University, Korea.
Ha
M S
MS
Kim
Y K
YK
D001932:Brain Neoplasms; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects*; D005456:Fluorescent Dyes; D016166:Free Radical Scavengers / Q000494:pharmacology*; D005910:Glioma; D005978:Glutathione / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008627:Mercuric Chloride / Q000633:toxicity*; D019335:Nitroarginine / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured; D014527:Uric Acid / Q000494:pharmacology
D005456:Fluorescent Dyes; D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D019335:Nitroarginine; D014527:Uric Acid; D019328:Buthionine Sulfoximine; D008627:Mercuric Chloride; D005978:Glutathione