Objective: To study the anti-fatigue effects of salidroside in mice. Methods: Totally 120 normal male Kunming mice were randomized into 5 groups (4 salidroside intervention groups and the control group) based on body weight. The control group was given distilled water and the 4 intervention groups were given various doses of salidroside (60, 180, 360, 720 mg/kg) for 15 consecutive days, respectively. The levels of lactate, serum urea nitrogen, muscle and liver glycogen, the longest swimming time and hemoglobin were determined before and after swimming test. Results: Different doses of salidroside significantly lengthened the swimming time and increased the contents of hemoglobin and muscle and liver glycogen, while reducing that of lactate in blood significantly compared with control group, especially in the 180 mg/kg salidroside group. Conclusion: Salidroside has noticeable anti-fatigue effect on mice. These effects were dose-dependent, and the strongest effect on most biomarkers was seen with an intermediate dose.