Protective effect of ebselen against hydrogen peroxide-induced cytotoxicity and DNA damage in HepG₂ cells

The protective effect of ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one), a selenoorganic compound, against hydrogen peroxide (H₂O₂)-induced cytotoxicity and DNA damage was investigated in a human hepatoma cell line, HepG₂. The inhibitory effect of H₂O₂ on cell growth was determined using the tetrazolium dye colorimetric test (MTT test), and the cytotoxicity and lipid peroxidation were estimated by lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation, respectively. DNA damage was detected using single cell gel electrophoresis (comet assay), and intracellular reactive oxygen species (ROS) formation was measured using a fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA). The results showed that H₂O₂ suppressed the growth of HepG₂ cells and the addition of ebselen significantly reduced the suppression. Furthermore, ebselen also displayed a dose-dependent reduction of LDH leakage and MDA formation in H₂O₂-treated cells. The results also demonstrate that ebselen was able to reduce the ROS formation and DNA damaging effect caused by H₂O₂ in a dose-dependent manner. These findings suggest that ebselen has a strong protective ability against the cytotoxicity and DNA damaging effect caused by reactive oxygen species. Copyright (C) 1999 Elsevier Science Inc.