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The response of orexin-A and brain-derived neurotrophic factor (BDNF) to exercise and diet are not fully understood. We examined the effect of acute aerobic exercise 2 h after the consumption of four meals on serum levels of orexin-A and BDNF in overweight men. We used a quasi-experimental and single-blinded crossover study in 16 overweight men (age 23 ± 6.1 y) who voluntarily participated in four sessions separated by 7d, consisting of 30-min of treadmill running (60–70% of maximum heart rate) after consuming one of four selected meals: Normal (Normal), High carbohydrate (HCHO), High protein (HP) and High-fat (HF) meals. Blood samples were taken after fasting, two hours after meal consumption (before exercise), and after exercise test. The results indicated that orexin-A and BDNF levels significantly increased after acute exercise test following HCHO (18.24 ± 25.06% and 13.38 ± 4.93%, respectively, P < 0.05), and HP (14.92 ± 19.02% and 9.23 ± 4.61%, respectively, P < 0.05): however, there was no significant increase after Normal meals (7.03 ± 35.51% and 6.66 ± 5.56%, respectively, P > 0.05), and HF meals (−0.43 ± 8.97% and −0.06 ± 4.01%, respectively, P > 0.05). When the four conditions were compared, between group comparisons demonstrated that orexin-A was significantly higher in HCHO (P ≤ 0.019) and HP meal (P ≤ 0.033) vs. HF meal after exercise. Further, BDNF was significantly higher after exercise in HCHO (P ≤ 0.001) and HP (P ≤ 0.001), and Normal (P ≤ 0.039) meals vs. HF meal. It appears that the exercise-induced increase in orexin-A and BDNF following caloric reduction is attenuated by HF meals in overweight men. [ABSTRACT FROM AUTHOR]