Siguan acupoints have been used to treat gastrointestinal symptoms in acupuncture practices for a long time. This study aimed to investigate the effects of Siguan acupuncture on gastrointestinal motility under accelerated conditions using a randomized, sham-acupuncture-controlled, crossover study. Twenty-one healthy male subjects were hospitalized and randomized into either a real acupuncture group (at Siguan acupoints) or a sham acupuncture group. Subjects were administered with mosapride citrate (15 mg a day) for 2 days starting 24 hours before the first acupuncture treatment. Immediately after the administration of radio markers, acupuncture treatment was conducted 4 times at 12-hour intervals. Gastrointestinal motility was assessed using radiograph distribution of the radio-markers located in the small intestine, ascending colon, transverse colon, descending colon, rectum, and outside the body immediately after the first acupuncture treatment and at 6, 12, 24, and 48 hours. After a 2-week washout period, the real acupuncture group in the first session was treated with sham acupuncture in the second session, and vice versa. Gastrointestinal motility was generally reduced in the real acupuncture group compared with the sham acupuncture group throughout the 4 different time points. A significant difference was observed at 24 hours following the first acupuncture treatment (P < 0.05).