Background: Following the SARS-CoV-2 pandemic in 2019, medical face masks were widely recommended for large numbers of people and for long periods of time. The effect of wearing surgical and N95 face masks during long-term high-intensity interval training on the cardiac autonomic system has not been reported. Methods: Eighteen healthy men were randomly divided into three groups (surgical mask, N95 mask, and no mask) and performed high-intensity interval training with (80 - 90%-heart rate max) intensity three times a week, three repetitions in a session for four weeks. Heart rate variability parameters were evaluated with an electrocardiogram device and Kubius software. The R-R interval was recorded for 5 minutes in a sitting position in the pre-and post-modified Bruce test before and after four weeks of interval training to extract parasympathetic (RMSSD, HF, SD1), sympathetic indices (LF, LF/HF) and RR mean. We used the two-way ANOVA test and the Bonferroni post hoc test. Results: In all groups, four-week functional HIIT exercises increased the resting parasympathetic indices (RMSSD, SD1) compared to baseline levels. This increase was more evident in the N95 mask group than in the surgical mask group. In addition, the five-minute recovery of the cardiac autonomic system after a modified Bruce test following functional HIIT exercise increased parasympathetic tone (RMSSD, HFn.u) in all groups compared to baseline levels. Therefore, a four-week adaptation to HIIT exercises leads to faster recovery of sympathetic indices (LFn.u, LF/HF) in the no mask group, as compared to the surgical and N95 groups. Despite the above, there was no significant difference in the recovery and resting values of cardiac autonomic system indices between the two types of surgical masks and N95 compared to the control group (no mask). Conclusions: Four weeks of high-intensity interval training using surgical and N95 face masks improved the cardiac autonomic nervous system. This data is important for recommending the use of face masks during physical activity.