Objectives The eye movement (EM) has been reported to play a role in enhancing the retrieval of episodic memories and reducing effects of fearful episodes in the past and worries for the futures. However, it is still unclear in the mechanism of EM in normal subjects. We examined the horizontal eye movement (HEM) effect using an aiding apparatus on mental health indices including negative and positive psychological factors, and psychophysiological measures such as heart rate variability and quantitative electroencepaholography (qEEG) in healthy subjects. Methods Twenty eight healthy subjects were recruited and randomly allocated into two groups : active HEM group and control group. The active HEM group conducted the HEM training with usual stress management audio-intervention using the apparatus inducing eye movement once a day for 14 days. The control group also conducted the same training once a day for 14 days, however, the saccadic eye movement was not included in this training. Psychological measurements, neurocognitive function tests, heart rate variability measurement and qEEG were conducted before and after the training in both groups. Results In the active HEM group, sleep status using Sleep Quality Scale (SQS) positive factors significantly increased after the training. By contrast, scores on the negative items of Psychological Well-Being Scale (PWBS), and negative items of the Life Orientation Test-Revised (LOT-R) were significantly decreased after the training. The percentage of delta amplitude (1-3 Hz) in qEEG significantly decreased after the HEM training. The percentage of alpha amplitude (8-12 Hz) significantly increased after HEM training. The change of delta amplitude in the active HEM group was positively correlated with the change of sleep satisfaction of Visual Analogue Scale (VAS), and the change of alpha amplitude was negatively correlated with depression of VAS, anxiety of VAS and Beck Anxiety Inventory (BAI). Conclusions The HEM training improved sleep quality and well-being, and sense of optimism. The HEM training also increased alpha amplitude and decreased delta amplitude in qEEG. The qEEG changes were well correlated with subjective improvement of mental health indices in healthy subjects. These results suggest some evidences that HEM training using the apparatus that induces EM would be helpful in improving subjective mental health in healthy subjects. Further study with larger samples size would be needed.