Heart rate variability (HRV) is the variation in time between each heartbeat. Increasing HRV may contribute to improving autonomic nervous system dysfunctions. Acupuncture stimulation through the vagus plexus in the ear is considered as a method that can improve HRV. In this pilot study, we examined 114 healthy volunteers at the Faculty of Traditional Medicine, University of Medicine and Pharmacy at Ho Chi Minh City, from January to May 2020. During a 20-minute interval, participants were stimulated two times at the acupoint in the left ear with Semen seed. The heart rate and HRV values were monitored before, during, and after acupressure every 5 minutes. When we compared the experimental group with the control group, HRV significantly increased in the stage of ear-stimulated acupressure compared with the stage before and after the auricular acupressure (
p
=
0.01
,
p
=
0.04
,
p
=
0.04
and
p
=
0.02
) and the difference was not statistically significant compared with the phase of nonstimulated (
p
=
0.15
,
p
=
0.28
). The changes in other values including SDNN (standard deviation of the average NN), RMSSD (root mean square of successive RR interval differences), LF (low-frequency power), and HF (high-frequency power) in all stages were not statistically significant
p
=
>
0.05
between groups. Based on the results, we can determine the increase in HRV when conducting auricular acupressure with stimulation at the heart acupoint on the left ear. This leads to a direction in further studies for clinical application for patients with autonomic nervous disorder.