Objective This study aims to investigate the effects of sleep interruption (SI) cycles on emotional behavior in ICR mice and establish a mouse model of comorbid anxiety and depression induced by sleep interruption. Methods 72 male ICR mice (4-5 weeks old) were randomly divided into a blank group and a model group based on 1, 2, and 3-week stress cycles. Except for the blank group, the model group was subjected to SI stress modeling for 1, 2, and 3 weeks, respectively. After modeling, emotional behaviors were evaluated using the open field test, elevated plus maze, light-dark box, marble burying test, and forced swim test. Serum corticosterone (CORT) levels were detected using ELISA. Results After 1 week of SI stress, the number of marbles buried by mice in the model group increased significantly compared to the blank group (P<0.05). After 2 weeks of stress, compared to the control group, the model group showed a significant decrease in the number of crossings in the light-dark box (P<0.05) and a significant increase in the number of marbles buried (P<0.01). After 3 weeks of stress, the model group showed a significant increase in the number of marbles buried (P<0.05), a significant decrease in the number of crossings in the light-dark box (P<0.05), and a significant increase in immobility time in the forced swim test (P<0.01). Conclusion ICR mice exhibited significant anxiety behaviors after 2 weeks of SI modeling and displayed significant anxiety and depressive behavior changes after 3 weeks. Three weeks of SI stress successfully established a model of comorbid anxiety and depression.