Naringenin and its glycoside naringin (naringenin-7-O-glycoside) belong to this series of flavonoids and were well-known strong antioxidant activity. This study was conducted to flavonoid constituents and antioxidant activity of Pleurotus ostreatus and Pleurotus cornucopiae. In order to determine active ingredient contents of Pleurotus ostreatus and Pleurotus cornucopiae, we were carried out total polyphenolic content (TPC) and flavonoid content (TFC) analyses. As a result, TPC (31.05 ± 0.03 g GAE/kg extract) and TFC (13.25 ± 1.24 g NE/kg extract) of Pleurotus cornucopiae were found significantly higher as compared to Pleurotus ostreatus. The IC50 values based on the DPPH (56.50 ± 0.73 μg/ml) and ABTS (86.53 ± 3.86 μg/ml) for Pleurotus cornucopiae were generally stronger showing potential antioxidant properties compared to Pleurotus ostreatus. In addition, naringin and naringenin content were analyzed by high-performance liquid chromatography analysis. A great amount of naringin and naringenin was found in Pleurotus cornucopiae. These results indicate that the Pleurotus cornucopiae can be used as an antioxidant therapeutic agent.