Objective To explore the effect of acupuncture on gastrocnemius fatigue. Methods 20 healthy volunteers were selected. Accordingto the rating of perceived exercise (RPE) table, when exercise-induced fatigue occurred, the volunteers were needled in PointChengjin for 15 minutes (acupuncture group). 4 weeks later, they just rest after exercise-induced fatigue occurred for the second time for 15minutes (non-acupuncture group). The data of surface electromyographic signal (sEMG) and heart rate were collected. Results The value oflow frequency/high frequency (LF/HF) of sEMG of both groups was higher after exercise. The acupuncture group decreased after acupuncture(P<0.05), the non-acupuncture group increased after rest (P<0.05). The heart rate of both groups increased significantly after exercise(P<0.01), and decreased after acupuncture and rest (P<0.05), and there was no significant difference in the decrement between 2 groups (P>0.05). Conclusion Acupuncture can relieve gastrocnemius fatigue.