Objective: To examine the effect of electroacupuncture (EA) stimulation on multiple physiological indices and to evaluate both local and systemic physiological responses induced by the stimulation. Methods: 15 healthy male college students participated in an experimental crossover study. They received two kinds of interventions: one with EA stimulation and one without EA stimulation on different days. Two disposable acupuncture needles were inserted at two traditional acupuncture points (ST36 and ST38), located along the anterior tibialis muscle. EA stimulation was administered for 10 min. Skin temperature (ST), skin blood flow (SBF) and muscle blood volume (MBV) were recorded near the stimulation sites, while the pupil diameter (PD) was measured before, during and after the interventions. Results: ST, SBF and MBV increased significantly following EA stimulation. PD of the right and left eyes decreased significantly following EA stimulation. There was a significant difference in ST responses between the groups (P=0.001). For SBF, MBV and PD, no significant differences were demonstrated between the groups. Conclusions: Our study showed that 10 min of EA stimulation increased ST, SBF and MBV, and decreased PD, compared to baseline, while no significant change was observed in the control group. This suggests that EA stimulation alters local blood flow and ST, and these responses are likely mediated via segmental spinal reflexes, supraspinal reflexes involving parasympathetic activation, and other mechanisms.