〔Abstract〕 Objective To establish a respiratory fluctuation evaluation method to evaluate the autonomic respiration status in the experiment of regulating respiratory function by cerebral nuclei. Methods Eighteen rats were randomly divided into blank group (N) (normal rats, respiratory function monitored by tracheal intubation), solitary tract nucleus (NTS) electrode implantation group (NTS-C) (normal rats, respiratory function monitored by tracheal intubation, concentric stimulation electrode implanted in NTS), and NTS electrical stimulation group (NTS-E) (normal rats, respiratory function monitored by tracheal intubation, concentric stimulation electrodes were implanted into NTS and received direct current stimulation), 6 rats in each group. Respiratory parameters were collected by endotracheal intubation and multi-channel physiological monitor, and respiratory function was evaluated by respiratory rate, tidal volume, maximum inspiratory volume and maximum expiratory volume. The breathing fluctuation was calculated according to the receiver operating characteristic curve (ROC) to evaluate the active breathing status. Results After NTS electrical stimulation, the mean blood pressure of group NTS-E was significantly decreased and the heart rate was significantly increased compared with that of group N, the differences were statistically significant (P < 0.01). There was no significant difference in classical respiratory parameters between groups after NTS stimulation (P > 0.05). After electrical stimulation of NTS, the frequency of respiratory enhancement wave in group NTS-E was significantly increased. Compared with group N, the respiratory fluctuation in group NTS-E and group NTS-C was higher than that in group N, and that in group NTS-E was higher than that in group NTS-C, the differences were statistically significant (P < 0.01). Conclusion The evaluation method of respiratory fluctuation can effectively evaluate the changes of active respiration, and can be used to monitor the electrical stimulation of respiratory related cerebral nuclei in rats by tracheal intubation.