title: Peroxide-induced cell death and lipid peroxidation in C6 glioma cells.
abstract: Peroxides are often used as models to induce oxidative damage in cells in vitro. The aim of the present study was to elucidate the role of lipid peroxidation in peroxide-induced cell death. To this end (i) the ability to induce lipid peroxidation in C6 rat astroglioma cells of hydrogen peroxide (H2O2), cumene hydroperoxide (CHP) and t-butyl hydroperoxide (t-BuOOH) (ii) the relation between peroxide-induced lipid peroxidation and cell death in terms of time and concentration dependency and (iii) the capability of the lipid peroxidation chain breaking alpha-tocopherol to prevent peroxide-induced lipid peroxidation and/or cell death were investigated. Lipid peroxidation was characterised by measuring thiobarbituric acid reactive substances (TBARS) and, by HPLC, malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) and hexanal. Within 2 h CHP, t-BuOOH and H2O2 induced cell death with EC50 values of 59+/-9 microM, 290+/-30 microM and 12+/-1.1 mM, respectively. CHP and t-BuOOH, but not H2O2 induced lipid peroxidation in C6 cells with EC50 values of 15+/-14 microM and 130+/-33 microM, respectively. The TBARS measured almost exclusively consisted of MDA. 4-HNE was mostly not detectable. The concentration of hexanal slightly increased with increasing concentrations of organic peroxides. Regarding time and concentration dependency lipid peroxidation preceded cell death. Pretreatment with alpha-tocopherol (10 microM, 24 h) prevented both, peroxide-induced lipid peroxidation and cell death. The results strongly indicate a major role of lipid peroxidation in the killing of C6 cells by organic peroxides but also that lipid peroxidation is not involved in H2O2 induced cell death.
authors.affiliation: Institute for Toxicology and Pharmacology for Natural Scientists, University Medical School Schleswig-Holstein, Campus Kiel, Brunswiker Strasse 10, 24105 Kiel, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Linden
authors.forename: Michael
authors.initials: M
authors.lastname: Gülden
authors.forename: Hans-Jörg
authors.initials: HJ
authors.lastname: Martin
authors.forename: Edmund
authors.initials: E
authors.lastname: Maser
authors.forename: Hasso
authors.initials: H
authors.lastname: Seibert
mesh_terms: D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001555:Benzene Derivatives / Q000633:toxicity; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D005910:Glioma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D006861:Hydrogen Peroxide / Q000633:toxicity; D015227:Lipid Peroxidation / Q000187:drug effects; D008315:Malondialdehyde / Q000378:metabolism; D016877:Oxidants / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D010545:Peroxides / Q000633:toxicity*; D051381:Rats; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D024502:alpha-Tocopherol / Q000494:pharmacology; D020122:tert-Butylhydroperoxide / Q000633:toxicity
chemical_list: D000447:Aldehydes; D000975:Antioxidants; D001555:Benzene Derivatives; D004338:Drug Combinations; D016877:Oxidants; D010545:Peroxides; D017392:Thiobarbituric Acid Reactive Substances; D008315:Malondialdehyde; D020122:tert-Butylhydroperoxide; C010463:n-hexanal; D006861:Hydrogen Peroxide; D024502:alpha-Tocopherol; C027576:4-hydroxy-2-nonenal; C007164:cumene hydroperoxide