The protective and prothetic effect of sea buckthorn seed extracts （SBSE） on oxidative damage in B16F10 was studied. Firstly， the oxidative damage model was established by treatment with H2O2. The evaluation of the damage model was done by the flow cytometry. Secondly， cells were treated by SBSE before or after the establishment of the damage model. The cell viability was measured to discuss the protective and prothetic effects of SBSE. Thirdly， the contents of glutathione peroxidase （GSH-Px）， superoxide dismutase （SOD）， catalase （CAT） and malondialdehyde （MDA） were studied. The results showed that the oxidative damage model was established by the treatment with H2O2（300 μg/mL） for 4 h. The flow cytometry showed that the percentage of living cells decreased from （91.61 ± 1.14）% to （49.77 ± 2.74）% （P<0.01）， the percentage of early apoptotic cells rose from （1.97 ± 0.34）% to （17.91 ± 3.55）%， and the late apoptosis rate increased to （21.24 ± 2.61）%. Besides， the late apoptosis rate reached more than 60% after 6 hours’ treatment， dramatically. Cells firstly treated by SBSE and then H2O2 were used to examine the cell viability and cellular antioxidant enzymes. After the pretreatment of 0.10 mg/mL SBSE， the cell viability was increased significantly， and the levels of cellular antioxidant enzymes were also increased （P<0.05）. Besides， the pretreatment of SBSE could decrease the release of MDA（P<0.05）. The prothetic effect of SBSE was detected when the damage model cells treated by SBSE of 0.10 mg/mL（P<0.05）. The treatment of SBSE on the model could increase the activities of GSH-Px， CAT and SOD （P<0.05， P<0.01， P<0.01）， and decrease the level of MDA（P<0.05）， significantly. In conclusion， SBSE showed protective and prothetic effects against H2O2 induced oxidative damage in B16F10 through increasing the activities of GSH-Px， CAT and SOD， and inhibiting MDA contents.