Evidence for the involvement of oxygen free radicals in the ethanol-induced late cellular injury in mouse myeloma cells.
In this study, we analysed the ethanol-induced long term cell injury on a general cell model (Sp2/0-Ag14 cell line). Cells were incubated in 1, 5, 10, 15 and 20% of ethanol (EtOH) for 5 min. After washing cell viability was tested by the Trypan Blue exclusion test in 5, 60 min, 4 and 24 h after EtOH exposure. Free radicals were monitored every 30 min by electron spin resonance (ESR) with alpha-phenyl-N-tert-butylnitrone (PBN) spin trapping technique. Scavenger compounds such as glutathione (GSH), dimethyl sulfoxide (DMSO) and 5,5-dimethyl-1-pyrroline N-oxide (DMPO) were applied for 24 h incubation after EtOH exposure. EtOH concentration dependently decreased the cell viability immediately after 5 min exposure, but with 4 and 24 h, a secondary cell destruction was found. Using ESR-spin trapping technique, an increased free radical activity could be detected. DMPO, DMSO and GSH significantly, but in different period protected the cells against free-radical induced cellular damage. EtOH produces an early (immediately after EtOH exposure) and a late (in about 4 h) cellular damage on Sp2/0-Ag14 cells. The oxygen free radicals can be detected in a short time after EtOH exposure, its biological effect manifested as a secondary cell destruction at 4 and 24 h. This phenomenon can be prevented by scavenger compounds.
Bódis
B
B
First Department of Medicine, University Medical School of Pécs, Hungary.
Karádi
O
O
Szabó
I
I
Németh
P
P
Belágyi
J
J
Mózsik
G
G
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D000431:Ethanol / Q000494:pharmacology*; D016166:Free Radical Scavengers / Q000494:pharmacology; D051379:Mice; D009101:Multiple Myeloma / Q000473:pathology*; D009994:Osmolar Concentration; D017382:Reactive Oxygen Species / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology
D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D000431:Ethanol