Background: This study compared the effect of moderate and high-intensity interval training on plasma levels of orexin-A, insulin, and insulin resistance in women with overweight or obesity. Methods: Thirty-six women were voluntarily selected and optionally assigned into high-intensity interval training (HIIT; BMI = 29.43 ± 3.16 kg/m2, n = 12), moderate-intensity interval training (MIIT; BMI = 27.63 ± 2.82 kg/m2, n = 12), and control groups (BMI = 28.77 ± 4.28 kg/m2, n = 12). The exercise program consisted of selected high-intensity and moderate-intensity exercises with 75 - 95% and 65 - 80% of the heart rate reserve of initial fitness level for 30 - 60 minutes each session and three times a week. Plasm levels of orexin-A and other biochemical parameters were measured pre-and post‐interventions. Results: Following exercise interventions, plasma levels of orexin-A improved (MIIT: 23.23% vs. HIIT: 23.27%). Also, glucose (MIIT: -8.83 vs. HIIT: -6.01%), insulin (MIIT: -20.02% vs. HIIT: -25.38%), insulin resistance index (MIIT: -8.37% vs. HIIT: -3.28%), and body fat percentage (MIIT: -6.46% vs. HIIT: -7.23%) reduced in both MIIT and HIIT groups, but between-group differences were not statistically significant (P ≥ 0.05). A significant increase in the cardiorespiratory endurance of participants was observed (MIIT: 10.10 vs. HIIT: 14.83%) in both HIIT and MIIT exercise groups (P ≤ 0.05). Conclusions: The findings showed that HIIT and MIIT could improve orexin and some cardio-metabolic health biomarkers in sedentary women with overweight/obesity.