Objectives: To investigate the different effects of Sa-Am acupuncture with Spleen Jung-gyuck and Spleen Seung-gyuck on the radial pulse in healthy subjects. Methods: Sixty healthy volunteers (30 males and 30 females) participated in this study. The participants were randomly divided into three groups: control (C), Sa-Am acupuncture with Spleen Jung-gyuck (SP+) and Sa-Am acupuncture with Spleen Seung-gyuck (SP-). The radial pulse was measured using a multi-step tonometry system before, and 0, 30 and 60 minutes after acupuncture. Results: The heart rate adjusted radial augmentation index (RAI/HR) and high-tensioned pulse area (W area) significantly increased in the SP- group compared to the C and SP+ groups in males. The systolic pulse period (T4) increased significantly in the SP- group compared to the C and SP+ groups in females. The RAI/HR had positive correlations with W area and T4. Conclusion: The effects of Sa-Am acupuncture with Spleen Jung-gyuck and Spleen Seung-gyuck are different. Sa-Am acupuncture with Spleen Seung-gyuck induces acute increases of radial pulse parameters related to arterial stiffness in healthy subjects.