Cornu cervi pantotrichum (CCP) is a well-known yang-invigorating agent used in traditional Chinese medicine that can nourish the blood, tonify qi, and invigorate bones and tendons with multifunctional bioactivities. However, evidence on the effects of CCP on exercise performance and physical fatigue is limited. We evaluated the potential beneficial effects of ethanolic extract from CCP on ergogenic and antifatigue functions following a physiological challenge. Male ICR mice from four groups (n=8 per group) were orally administered CCP for 14 days at 0, 2054, and 4108 mg/kg/day, and were respectively designated as the vehicle, CCP-1X, and CCP-2X groups. The physical performance and antifatigue function were evaluated using forelimb grip strength and exhaustive swimming time as well as serum levels of lactate, ammonia, glucose, and creatine kinase after a 15-min swimming exercise. The results indicated that CCP-1X supplementation significantly improved grip strength; reduced fatigue-associated biochemical indices, including lactate and ammonia levels; and ameliorated skeletal muscle injury induced by acute exercise challenge. A trend analysis revealed that CCP supplementation significantly increased grip strength and dose-dependently reduced serum alkaline phosphatase, uric acid, triacylglycerol, and glucose levels in healthy mice. Therefore, CCP is a potential agent with an antifatigue pharmacological effect.