Objectives: The objective of this study was to assess the effects of acupuncture applied at PC6 and SP4 points on heart rate variability (HRV) in normal subjects under mental stress Methods: 24 healthy male subjects were recruited and randomized to a treatment group (12 subjects) or a control group (12 subjects). Both groups performed a mental arithmetic stress test and then acupuncture needles were inserted on PC6 and SP4 for the study group. In the control group, they took sham acupuncture as a control. Heart rate variability (HRV) was measured during the whole study period by FM-150 (digital Holter ECG recorder). Results: In the both groups, low frequency (LF) power and ratio of low frequency to high frequency (LF/HF) was significantly increased after mental stress. Acupuncture induced significant decrease in low frequency (LF) power and ratio of low frequency to high frequency (LF/HF) which were increased after mental stress test. But sham acupuncture induced no changes. Results: In both groups, low frequency (LF) power and ratio of low frequency to high frequency (LF/HF) significantly increased after mental stress. Acupuncture induced significant decrease in low frequency (LF) power and ratio of low frequency to high frequency (LF/HF) which increased after mental stress test. In contrast, sham acupuncture induced no changes. Conclusions: Acupuncture on PC6 and SP4 could be useful to decrease sympathetic activity and balance autonomic nervous system for those who are under stress.