This study was aimed to compare the effects of acupuncture stimulation on different acupoints, which included LR14-Qimen, ST25-Tianshu, GB34-Yanglingquanand CO11-pancreas and gallbladder (auricular vagus nerve intensively distributed areas) on gallbladder pressure and Oddi sphincter function of guinea pig, in order to discuss acupuncture effect on different acupoints in the regulation of motor function of extrahepatic biliary system. A total of 20 adult guinea pigs of both sexes were randomly divided into two groups, with 10 in each group. The detection was made on changes of Oddi sphincter electromyography and gallbladder pressure before and after acupuncture. A double-hook electrode was implanted into the Oddi sphincter for the detection of Oddi sphincter electromyography. And one small artificial balloon was put into the gallbladder to measure the intrabiliary pressure. After the guinea pig was anesthetized, changes of gallbladder pressure as well as the electromyography frequency and amplitude index of Oddi sphincter of the normal guinea pig were recorded. When the guinea pig was stabilized, acupuncture was given on LR14, ST25, GB34 and CO11 for 1 min. Observations were made on acupuncture effects of gallbladder pressure and Oddi sphincter function of guinea pig. The results showed that after acupuncture on LR14 and ST25, the electromyography frequency of Oddi sphincter was obviously increased compared with before acupuncture (frequency:P < 0.01, amplitude:P < 0.05); the gallbladder pressure was obviously decreased (P < 0.05). After acupuncture on CO11 and GB34, the electromyography frequency and amplitude index of Oddi sphincter were obviously decreased compared with before acupuncture (P < 0.05); the gallbladder pressure was obviously increased (P < 0.01). It was concluded that acupuncture on LR14 and ST25 can obviously inhibit the movement of gallbladder and promote the myoelectricity release of Oddi sphincter. Acupuncturing on CO11 and GB34 can apparently promote the shrinking movement of gallbladder and inhibit the myoelectricity release of Oddi sphincter.