Acylation of anthocyanin with fatty acid has been extensively applied for the improvement of their lipophilicity and stability. However, their functionalities are not fully investigated. This work prepared cyanidin-3-O-gluco-side-lauric acid ester (C3G-LA) and investigated its protective effect against oxidative stress in H₂O₂-induced human normal liver (LO2) cells. The results indicated that C3G-LA possessed notable antioxidant activity and better protective effect than its anthocyanin precursor cyanidin-3-O-glucoside (C3G). It protected the cells from H₂O₂-induced cell death and apoptosis by suppressing the overproduction of reactive oxygen species and malondialdehyde, restoring superoxide dismutase activity and ameliorating mitochondrial dysfunction and membrane damage. The increase in the expression levels of phosphorylated Akt, Nrf2, HO-1 and NQO1 further indicated that the antioxidant protective effect of C3G-LA involved the PI3K/Akt-mediated activation of the Nrf2-HO-1/NQO1 pathway. The findings indicated that, in addition to improve lipophilicity and stability, acylation of anthocyanin can enhance their anti-oxidative stress activity.