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 moderate-intensity 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.