To investigate the protective effect of hawthorn kernel and screen its active fractions on oxidative damaged cells, providing supports for its applications, the oxidative damaged HaCaT cells and GES-1 cells were induced by H2O2. MTT assay and biochemical kit were used to detect cell viability and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the oxidation-damaged cells. The chemical components analysis method was used to study the chemical components of the active fractions. The results showed that hawthorn kernel ethanol extract (EE) and its derived fractions got from ethyl acetate (EAF) and water (WF) had protective activity on oxidative damaged cells. 25 μg/mL of EAF significantly increased the survival rate of oxidation-damaged HaCaT cells 21.23%, the levels of SOD in cells were also increased 9.09%. 25 μg/mL of WF significantly improved HaCaT and GES-1 cells survival rate 16.20% and 23.61%, increased the levels of SOD 16.62% and 17.38%, increased the levels of GSH-Px in cells 1.22 and 2.50%, respectively. The chemical components of ethyl acetate fraction were sugar (32.73%), flavonoids (15.70%), lignans (8.09%) etc. The water fraction contained sugar (59.59%) and flavonoids (1.14%) etc. It was suggested that WF can protect HaCaT and GES-1 cells from H2O2 induced oxidative damage, and its main component is sugar. The mechanism of WF may be related to the regulation of intracellular antioxidant enzyme levels, which has a good development and application values.