Introduction: Scientific studies confirm that unhealthy habits play an important role in the development of various disorders in all age groups. The field of medicine is a challenging period for students of this field that causes unwanted changes in their health habits and lifestyle. Objectives: The aim of this study is the evaluate and compare the lifestyle among first and last year medical students of Isfahan university of medical sciences. Patients and Methods: This study is a descriptive-analytic study that was conducted in two groups of first year and last year medical students in Isfahan university in academic year 2019-2020. The learning styles questionnaire (LSQ) is used in this study. It consists of 70 questions and measures lifestyle in 10 domains. This questionnaire is based on 6-point Likert scale and each question receives at least one point and maximum 6 points. Results were obtained by SPSS software version 22 and independent t-test and chi-square test. Results: The age range of first year students was 18-24 years with a mean of 19.5 years and the age range of last year students was between 24 and 27 years with a mean of 25 years, which showed a significant difference (P<0.05; independent t test). There was no significant difference between the two groups in gender distribution and living status however, there was a significant difference between the two groups in marital status distribution. The mean scores of lifestyle scores in physical health dimensions and sport and physical activity in first year students were significantly higher than last year students since, the mean score of other aspects of life style was not significantly different between Junior and senior students. Conclusion: Overall, the mean scores of lifestyle scores in physical health dimensions and sport and physical activity in first year students were significantly higher than last year students however, the mean score of other aspects of life style was not significantly different between freshmen and senior students.