Protective effect of hydroxytyrosol against acrylamide-induced cytotoxicity and DNA damage in HepG2 cells

The chemoprotective effect of hydroxytyrosol (HT) against acrylamide (AA)-induced cytotoxicity and DNA damage was investigated in a human hepatoma cell line, HepG2. The cytotoxicity was estimated by methyl thiazol tetrazolium bromide (MTT) assay. The comet assay was used to monitor DNA damage. The intracellular reactive oxygen species (ROS) formation and the level of oxidative DNA damage were estimated by using 2,7-dichlorofluorescein diacetate (DCFH-DA) as a fluorescent probe and by using immunocytochemistry analysis of 8-hydroxydeoxyguanosine (8-OHdG). Intracellular glutathione (GSH) level was estimated by fluorometric methods. The results showed that HT significantly reduced the cytotoxicity, DNA damage, intracellular ROS formation and 8-OHdG level caused by AA in a concentration-dependent manner. It was also found that HT concentration-dependently attenuated GSH depletion in HepG2 cells treated with 10 mM AA. These findings suggest that HT has a strong protective ability against the cytotoxicity and DNA damage caused by AA.