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 g eneral 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 t ested 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 reson ance (ESR) with alpha-phenyl-N-tert-butylnitrone (PBN) spin trapping techni que. Scavenger compounds such as glutathione (GSH), dimethyl sulfoxide (DMS O) and 5,5-dimethyl-1-pyrroline N-oxide (DMPO) were applied for 24 h incuba tion after EtOH exposure. EtOH concentration dependently decreased the cell viability immediately after 5 min exposure, but with 4 and 24 h, a seconda ry cell destruction was found. Using ESR-spin trapping technique, an increa sed free radical activity could be detected. DMPO, DMSO and GSH significant ly, but in different period protected the cells against free-radical induce d 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 biol ogical effect manifested as a secondary cell destruction at 4 and 24 h. Thi s phenomenon can be prevented by scavenger compounds. (C) 2000 Elsevier Sci ence Ltd. Published by Editions scientifiques et medicales Elsevier SAS.