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This study investigated the protective effects of ampelopsin on H2O2-induced oxidative injury in porcine kidney epithelial cell line, PK-15; cells were pretreated with medium containing 0, 15, 30, and 60 μg mL-1 ampelopsin, respectively, for 1 h prior to the addition of 100 μmol L-1 H2O2· After 2 h, the cell viability, intracellular superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity, cellular malondialdehyde (MDA) content, and DNA strand breakage were determined. Results showed that ampelopsin pretreatment did not modify the SOD and GSH-Px inactivation but significantly decreased MDA production; except for the medium dose (30 μg mL-1), all the tested concentrations of ampelopsin did not significantly inhibit cell viability reduction and gave no influence on DNA damage. The results suggested that all the tested doses of ampelopsin pretreatment before H2O2 addition can protect PK-15 cell from H2O2-induced lipid peroxidation, which might not be mediated by the SOD and GSH-Px responsees. The appropriate dose of ampelopsin pretreatment prevents PK-15 cell from DNA damage and cell viability reduction.