According to a previous study, YGDEY from tilapia fish skin gelatin hydrolysates hasstrong free radical scavenging activity. In the present study, the protective effect of YGDEY againstoxidative stress induced by ethanol in HepG2 cells was investigated. First, cells were incubatedwith YGDEY (10, 20, 50, and 100 μM) to assess cytotoxicity, and there was no significant change incell viability. Next, it was established that YGDEY decreased the production of reactive oxygenspecies (ROS). Western blot results indicated that YGDEY increased the levels of superoxidedismutase (SOD) and glutathione (GSH) and decreased the expression ofgamma-glutamyltransferase (GGT) in HepG2 cells. It was then revealed that YGDEY markedlyreduced the expressions of bax and cleaved-caspase-3 (c-caspase-3); inhibited phosphorylation ofAkt, IκB-α, p65, and p38; and increased the level of bcl-2. Moreover, the comet assay showed thatYGDEY effectively decreased the amount of ethanol-induced DNA damage. Thus, YGDEYprotected HepG2 cells from alcohol-induced injury by inhibiting oxidative stress, and this may beassociated with the Akt/nuclear factor-κB (NF-κB)/mitogen-activated protein kinase (MAPK) signaltransduction pathways. These results demonstrate that YGDEY from tilapia fish skin gelatinhydrolysates protects HepG2 cells from oxidative stress, making it a potential functional foodingredient.