Acupuncture is commonly performed at acupoints. No comparisons of quantitative physiological alterations in the brachial artery (BA) induced by the stimulation of different acupoints in the lower limbs have been performed in humans. Therefore, we investigated changes in blood flow volume (BFV) in the BA as an indicator of the physiological effects induced by stimulation at 3 points. Seventy-five healthy participants aged 33 ± 9 years (mean ± SD) were enrolled and randomly assigned to 3 groups; they received stimulation at 3 different points located on the lower limbs: ST36, LR3, and a non-acupoint. Stimulation was performed bilaterally with manual rotation of the needles. Using ultrasonography, BFV was measured continuously from rest to 180 seconds after stimulation. LR3 stimulation significantly increased BFV compared to that before needle insertion. Meanwhile, stimulation at ST36 and the non-acupoint significantly decreased BFV compared to that before needle insertion. Stimulation at LR3 elicited a significant increase in BFV compared to that at ST36 and the non-acupoint. The results suggest that the stimulation of different points on the lower limbs causes distinct physiological effects on BFV in the BA.