The aim of this study was to assay changes in blood biochemical parameters that resulted from exercise-induced muscle fatigue in horses participating in the two races (1,250 and 1,400 meters). Six male Arabian horses (3 to 6 years old) were used in this study. Blood samples were collected at time intervals including 1 hour before the race, immediately after the race, 1 and 24 hours after the end of race. Catalase (CAT), superoxide dismutase, glutathione peroxidase (GPX) activities, the blood level of malondialdehyde, protein carbonyl, and total antioxidant capacity (TAC) were measured, as well as muscle damage biomarkers including aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine kinase (CK) activities, and myoglobin were measured. The results showed that CAT activity and plasma TAC in the horse increased immediately after the race and then gradually decreased. The highest GPX activity in red blood cells was recorded 1 hour before the start of the race. Superoxide dismutase showed an incremental pattern after the race. Immediately after the race, there was a significant increase in the plasma levels of AST, which continued until 1 hour after the race. The activity of LDH and CK reached its highest value 1 hour after the race. According to our findings, it can be concluded that the horses were tired and antioxidant enzymes altered under fatigue conditions. Muscle damage biomarkers have increased, but these increases were in their natural ranges and did not indicate muscle damage in horses.