title: Comparative impacts of glutathione peroxidase-1 gene knockout on oxidative stress induced by reactive oxygen and nitrogen species in mouse hepatocytes.
abstract: Selenium-dependent glutathione peroxidase-1 (GPX1) protects against reactive-oxygen-species (ROS)-induced oxidative stress in vivo, but its role in coping with reactive nitrogen species (RNS) is unclear. Our objective was to compare the protection of GPX1 against cytotoxicity of superoxide generator diquat (DQ), NO donor S-nitroso-N-acetyl-penicillamine (SNAP) and peroxynitrite generator 3-morpholinosydnonimine (SIN-1). Primary hepatocytes were isolated from GPX1-knockout (KO) and wild-type (WT) mice and cultured in complete Williams's medium E with various levels of these agents alone or in combination for up to 12 h. While the KO cells were more susceptible to cell death, DNA fragmentation and protein carbonyl formation induced by 0.25-1 mM DQ, these cells were as tolerant as the WT cells to cytotoxicity of 0.1-1 mM SNAP or 0.5-2 mM SIN-1. Treating cells with SNAP (0.1 or 0.25 mM) in addition to DQ produced synergistic cytotoxicity that minimized differences in apoptotic cell death and oxidative injuries between the KO and WT cells. Less protein nitrotyrosine was induced by 0.05-0.5 mM DQ+0.25 mM SNAP in the KO than in the WT cells. Total GPX activity in the WT cells was reduced by 65 and 25% by 0.5 mM DQ+0.1 mM SNAP and 0.5 mM DQ, respectively. Decreases in Cu,Zn-superoxide dismutase (SOD) activity and increases in Mn-SOD activity in response to DQ or DQ+SNAP were greater in the KO cells than in the WT cells. In conclusion, GPX1 was more effective in protecting hepatocytes against oxidative injuries mediated by ROS alone than by ROS and RNS together. Knockout of GPX1 did not enhance cell susceptibility to RNS-associated cytotoxicity. Instead, it attenuated protein nitration induced by DQ+SNAP.
authors.affiliation: Department of Animal Science, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Fu
authors.forename: J M
authors.initials: JM
authors.lastname: Porres
authors.forename: X G
authors.initials: XG
authors.lastname: Lei
mesh_terms: D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D053938:DNA Fragmentation; D004178:Diquat / Q000494:pharmacology; D004347:Drug Interactions; D005979:Glutathione Peroxidase / Q000235:genetics* / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006540:Herbicides / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D008981:Molsidomine / Q000031:analogs & derivatives* / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009584:Nitrogen / Q000737:chemistry / Q000378:metabolism*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000378:metabolism; D000094765:Glutathione Peroxidase GPX1
chemical_list: D006540:Herbicides; D020030:Nitric Oxide Donors; D017382:Reactive Oxygen Species; C002385:linsidomine; D026423:S-Nitroso-N-Acetylpenicillamine; D004178:Diquat; D008981:Molsidomine; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D009584:Nitrogen; D000094765:Glutathione Peroxidase GPX1; C000722068:Gpx1 protein, mouse