Background and purpose: Adipogenesis is the conversion of the pre-adipocytes cells into mature fat cells, which leads to the fat accumulation, overweight and obesity. The purpose of this research was to study the effect of eight weeks of aerobic training followed by twelve weeks of high-fat diet on expression of Pref-1, KLF2, GATA3, GATA2 and PPARγ genes in the visceral adipose tissue of male Wistar rats. Materials and methods: In this experimental research, the twenty male Wistar rats (age of eight weeks and initial weight of 220 ± 20 g) were randomly divided into standard diet (n=5) and high-fat diet (n=15) groups. At the end of the twelfth week and after the induction of obesity, the rats in high-fat diet group were randomly divided into three groups: high-fat diet control, high-fat diet aerobic exercise and normal diet aerobic exercise. Aerobic exercise program based on the principle of gradual overload was performed for eight weeks and five sessions in per week. 48 hours after the last training session, rats were anesthetized by intraperitoneal injection of ketamine and xylazine. Then the blood sample was taken directly from the heart and the visceral fat tissue was also extracted. The extracted sample was washed in PBS and stored at -80°C. The expression levels of Pref-1, KLF2, GATA3, GATA2 and PPARγ genes were measured by Real-time PCR method. the one-way analysis of variance (ANOVA) with Tukey's post-hoc test was used to analyze the data. The data were analyzed at a significance level of 0.5 by SPSS-23 software. Results: The results of the present study showed that a high-fat diet led to a significant decrease in the expression of Pref-1, KLF2, GATA3, GATA2, and a significant increase in the PPARγ gene (P < 0.05), while eight weeks of aerobic exercise increased significantly the expression of Pref-1, KLF2 , GATA3, GATA2 genes but PPARγ gene expression decreased significantly due to the aerobic exercise (P< 0.05). Conclusion: Aerobic exercise can increase the expression of adipogenesis inhibiting genes and prevent the accumulation of body fat and the development of obesity and related diseases.