Glutathione consumption and glutathione peroxidase inactivation in fibroblast cell lines by 4-hydroxy-2-nonenal.
Treatment of cultured fibroblasts, designated HA1 cells, with 4-hydroxy-2-nonenal (4HNE) in doses up to 50 nmol/10(6) cells for 3 h results in dose-dependent cytotoxicity measured by clonogenic cell survival with 50% cytotoxicity achieved at 32 nmol 4HNE/10(6) cells. 4HNE treatment also resulted in dose-dependent reduction of cellular glutathione (GSH) content and loss of glutathione peroxidase (GPx) activity at 4HNE doses greater than 15 nmol/10(6) cells. By comparison, a 95% oxygen-resistant variant of HA1 cells, designated O2R95 cells, and a hydrogen peroxide-resistant variant of HA1 cells, designated OC14 cells, were found resistant to 4HNE cytotoxicity requiring 54 nmol 4HNE/10(6) cells and 75 nmol 4HNE/10(6) cells, respectively, for 50% cytotoxicity. In O2R95 cells, dose-dependent decreases were seen in GSH levels and GPx activity. In OC14 cells, however, any reduction in cellular GSH levels required doses of 4HNE greater than 30 nmol/10(6) cells, and GPx activity remained unchanged. No changes were seen in glutathione-S-transferase activity in any of the cell lines at any dose tested. These data indicate a correlation between glutathione modification, in a manner that prevents its recycling, the ability to inactivate enzymes with active site selenocysteine residues and the cytotoxicity of alpha, beta-unsaturated aldehydes such as 4HNE.
Kinter
M
M
Department of Microbiology, University of Virginia Health Sciences Center, Charlottesville 22908, USA.
Roberts
R J
RJ
D000447:Aldehydes / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D002999:Clone Cells; D003432:Cross-Linking Reagents; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts; D005978:Glutathione / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors*; D007700:Kinetics
D000447:Aldehydes; D003432:Cross-Linking Reagents; D005979:Glutathione Peroxidase; D005978:Glutathione; C027576:4-hydroxy-2-nonenal