Background: Obesity can cause leptin and insulin resistance which will affect the production of GnRH and FSH. One of risks factor of obesity is high calorie intake chronicaly. Exercise plays a role in the regulation of blood calorie and body weight. However, the effect of exercise on subjects with a high-calorie diet on FSH is unknown. This study aims to determine the effect of a high calorie diet with moderate intensity interval training (MIIT) on FSH level in female rats.Method: Sample was 27 female rats (Rattus norvegicus) wistar strain were divided into 3 groups. The negative control group (standard diet), the positive control group (high calorie diet), and the MIIT group (high calorie diet and moderateintensity exercise with intervals). Measurement of FSH using the ELISA method after treatment for 4 weeks. Result: Statistical analysis showed that there was a significant difference in FSH level with a value (p = 0.001). The mean FSH was 25.32 mIU/ml ± 7.89 in the negative control group, 19.71 mIU/ml ± 6.55 in the positive control group, and 12.12 mIU/ml ± 3.64 in the MIIT group. The decrease FSH in the positive control and MIIT group due to a high calorie diet caused insulin and leptin resistance which affected GnRH. Conclusion: Moderate interval training decreases the FSH level may due to the level of physical stress.