The hypothesis was that stress in horses getting to a vibrating plate could be reduced within seven days. Based on this hypothesis, the aim of the study was to analyze the heart rate variability parameters of horses during their acclimation to the vibrating plate. The study involved 10 warmblood horses aged 9-21 years. The study was conducted on a Technohorse vibrating plate (platform). The experiment was planned for seven consecutive days. During the first three days, the horses were accustomed to entering the space containing the vibrating plate and to being placed on the on the device itself. During the next four days, the horses were placed on the vibrating plate, and then a three-minute program was initiated. Heart rate variability (HRV) parameters were also measured during the experiment. Based on the obtained results, it can be concluded that the assumed seven-day period is sufficient to prepare the horses for relaxation treatments on the vibrating plate. The observed ranges of heart rate variability parameters illustrate clear differences between the first and subsequent measurements, indicating lower emotional arousal of the horses on subsequent days of their acclimation to vibrotherapy. Activating the platform itself did not affect the activity of the horses’ autonomic nervous system when comparing the first and last activation of the device during the experiment. The heart rate (bpm) parameter, may be useful in assessing the emotional state, which may have significant application implications in terms of the use of vibrations in the regeneration of the horse’s body.