Objective To investigate the anti-fatigue effect of hairy antler composition and its mechanism. Methods NIH mice and SD rats were randomly divided into a blank control group, Yihua capsule group (600 mg / kg), and three dosage groups of hairy antler composition ( 135, 270, and 810 mg / kg). The Yihua capsule, hairy antler composition, at various concentrations or water were administered orally to the corresponding groups once a day for 30 days. Serum BUN and blood lactic acid after swimming, loaded swimming time, and hepatic glycogen content were measured at 30 min after the last administration. Results The hairy antler composition significantly increased the loaded swimming time and the hepatic glycogen content in mice (P<0. 05) and decreased blood lactic acid (P<0. 05). Conclusions The hairy antler composition has an anti-fatigue effect.