The purpose of this study was to evaluate the effects of aqueous extract from Atractylodes macrocephala (AME) on the physical fatigue. A total of 48 mice were randomly divided into four groups of 12 mice each: control (C), low-dose AME-treated (LAT), middle-dose AME-treated (MAT) and high-dose AME-treated (HAT) groups. The mice in the treated groups received AME (50, 100 and 200 mg/kg, respectively) intragastrically (ig) and the mice in the control group received distilled water ig. After 30 days, a forced swimming test was performed and the biochemical parameters related to fatigue were examined. The results suggested that AME could extend the swimming time to exhaustion of the mice, as well as increase the liver and muscle glycogen contents, and decrease the blood lactate and blood urea nitrogen levels. These finding indicates that AME had anti-fatigue effects.