This study compared the effects of distilled Rehmannia glutinosa, Wild Ginseng and Astragali Radix pharmacopuncture on the autonomic nervous system and heart rate variability. The purpose of the trial was to observe the influence distilled Astragali Radix, Wild Ginseng and Rehmannia glutinosa pharmacopuncture have on the autonomic nervous system. 120 healthy male volunteers were divided into four groups, which consisted of three experimental groups and a control group. This study was a randomized, placebo-controlled, double-blind clinical trial. Volunteers in experimental groups were underwent pharmacopuncture at GB21 (Kyonjong), and volunteers in the control group were injected with normal saline at GB21 (Kyonjong). Heart rate variability was measured seven times: before and after injection, every 5 minutes for 30 minutes. The result was distilled Rehmannia glutinosa, Wild Ginseng and Astragali Radix pharmacopuncture in healthy adult males tended to activate the autonomic nervous system, particularly the sympathetic nervous system.