To investigate the effect of electroacupuncture (EA) at different acupoint combination on intestinal motility and expression of inflammatory factors IL-6 and TNF-α in intestinal muscle layer in mice with postoperative ileus (POI), so as to provide reference of acupoint combination of clinical EA intervention for POI. A total of 78 SPF-grade male C57BL/6 mice aged 8-12 weeks were randomly divided into a sham operation group, a model group, a ST 36+ non-acupoint group, a ST 36+ ST 25 group, a ST 36+ CV 4 group and a ST 36+ CV 12 group, 13 mice in each group. The model of POI was established by intestinal manipulation. The mice in each acupuncture group were intervened with HANS-200A acupoint nerve stimulator within 3 hours after modeling, continuous wave, frequency of 10 Hz, intensity of 1 mA, for 20 min. The EA was given once. 22.5 h after model establishment, 7 mice in each group were intragastrically administered with fluorescein isothiocyanate-conjugated (FITC-dextran). 1.5 h after intragastric administration, intestinal motility was evaluated. 24 h after modeling, the expression of IL-6 and TNF-α mRNA in intestinal muscle layer was detected by real-time fluorescence quantitative PCR and spleen index was calculated. Compared with the sham operation group, the intestinal motility 24 h after modeling in the model group was significantly decreased (P<0.01), and the expression of IL-6 and TNF-α mRNA in intestinal muscle layer was increased (P<0.01). Compared with the model group, the intestinal motility 24 h after modeling in the ST 36+ non-acupoint group and the ST 36+ ST 25 group was increased (P<0.01), and the expression of IL-6 and TNF-α mRNA in intestinal muscle layer was decreased (P<0.01, P<0.05), and the expression of TNF-α mRNA in intestinal muscle layer in the ST 36+ CV 4 group was also decreased (P<0.05), but there was no significant difference in the ST 36+ CV 12 group and the ST 36+ CV 4 group (P>0.05). There was no significant difference in spleen index among the groups (P>0.05). EA at "Zusanli" (ST 36) or "Zusanli" (ST 36) combined with "Tianshu" (ST 25) can improve intestinal motility in mice with POI, and the mechanism may be related to the improvement of local inflammatory reaction. In addition, abdominal acupoints should be selected carefully.