The chemoprotective effect of hydroxytyrosol (HT) against UVB-induced DNA damage was investigated in a human skin keratinocyte cell line, HaCaT. The comet assay was used to monitor DNA strand breaks. Intracellular reactive oxygen species (ROS) formation was measured by flow cytometry using 2,7-dichlorofluorescein diacetate (DCFH-DA). The levels of oxidatively generated damage to DNA were estimated by immunocytochemistry analysis of 8-hydroxy-2'-deoxyguanosine (8-OHdG). The protein expression of p53 and NF-kappaB was estimated by western blotting. The results showed that HT significantly reduced the DNA strand breaks caused by UVB. It was also found that HT reduced intracellular ROS formation and 8-OHdG level caused by UVB. Furthermore, HT attenuated the expression of p53 and NF-kappaB in a concentration-dependent manner. These results strongly suggest that HT has a significant protective ability against UVB-induced DNA damage and that oxidative stress plays an important part in it.
(c) 2009 John Wiley & Sons, Ltd.