Acupuncture has been increasingly used in the treatment of muscle damage associated with sports activities. However, studies on the immediate effects of one-time acupuncture on the muscles of athletes are clearly lacking. Thus, this study aimed to examine the effects of acupuncture therapy on the maximal voluntary isometric contraction (MVIC) electromyography (EMG) of the rectus femoris and tibialis anterior muscles. This study was conducted among 20 healthy male college students who had no musculoskeletal disease. The participants were subjected to 3 different experimental conditions and subsequently grouped based on these conditions: real acupuncture, sham acupuncture, and control. A 7-day washout period was implemented to avoid any transient effects on the physiological and psychological conditions of the participants. Subsequently, an electromyogram patch was attached on the most developed area in the middle of the origin and insertion of the rectus femoris and tibialis anterior muscles. The percent MVIC, which was used to standardize the signal from the electromyogram, was determined, and the maximal value from the MVIC of the rectus femoris and tibialis anterior muscles was measured. The MVIC EMG activities of both femoris (F = 6.633, p = 0.003) and tibialis anterior (F = 5.216, p = 0.008) muscles were significantly different among all groups. Accordingly, the results of a posthoc test showed that the real acupuncture group had higher MVIC EMG activities in the femoris (p = 0.002) and tibialis anterior (p = 0.006) muscles compared with the control group. These results suggest that treatment with real acupuncture resulted in significantly higher MVIC EMG activities of the rectus femoris and tibialis anterior muscles than the other treatments. Hence, acupuncture may be helpful in the improvement of muscle strength among athletes in the physical fitness field.