The aim of the present study was to investigate the effect of electroacupuncture (EA) at the Zusanli (ST36) acupoint on somatostatin mRNA expression in the rat brainstem. A total of 96 Sprague-Dawley rats were randomly assigned to a control (n = 24), restraint (n = 24), pseudopoint-EA (n = 24), or Zusanli-EA (n = 24) group. The rats received stimulation for 30 min at 2, 4, 6, and 8 h after stimulation. Brainstem samples (n = 6 for each group at each time point) were collected, and somatostatin expression was assessed by reverse transcription polymerase chain reaction, northern blot, and immunohistochemical staining. Somatostatin mRNA expression was inhibited at 2 h after EA stimulation, but an increase was seen at 6 and 8 h in the Zusanli-EA group compared to the three control groups. With respect to immunohistochemical staining, the number of somatostatin-positive cells in the brainstem was increased in the Zusanli-EA group at 6 and 8 h after stimulation compared to the 2- and 4-h time point. These results indicate that Zusanli-EA increased somatostatin mRNA and protein expression in the brainstem. Somatostatin may therefore be involved in some of the analgesic effects of Zusanli-EA.