Objective: To investigate the use of mobile fitness application (APP) and healthy lifestyles of college students, so as to provide a theoretical basis for improving the health quality of college students and promoting the national scientific fitness. Methods: From August to October 2023, 591 college students from three universities in Jinan and Weifang Cities, Shandong Province were selected by stratified cluster random sampling method, and the online questionnaire was conducted on the mobile fitness APP use behavior and healthy lifestyle. The ttest and analysis of variance were used to assess the differences in healthy lifestyle of college students, and stratified linear regression was used to analyze the effect of mobile fitness APP use on healthy lifestyle. Results: The utilization rate of mobile fitness APP among college students was 82.57% (n=488), and the average score of healthy lifestyles was (3.49±0.66). The differences in students healthy lifestyle scores between different residences, grades, majors, monthly living expenses, annual personal visits, and family medical expenses were statistically significant (t/F=23.82, 4.87, 6.07, 10.17, 3.79, 16.92, P<0.05). The results of stratified regression analysis showed that the overall evaluation of mobile fitness APP, and the recommendation intention of mobile fitness phone APP were the main related factors of healthy lifestyles among college students (β=0.47, 0.06, P<0.05). Conclusions College students have a high degree of use of the mobile fitness APP, and the mobile fitness APP is positively associated with a healthy lifestyle.It should increase the publicity and promotion of mobile fitness APP to promote college students to develop a healthy lifestyle.