Abstract
The traditional horse industry has undergone a remarkable evolution, with horse racing emerging as a prominent and pivotal economic driver within the sector. Among the various breeds, Mongolian horses, renowned for their exceptional endurance and speed, occupy a significant position in the horse industry. To investigate their homeostasis mechanisms during and after a 20 km endurance exercise and identify novel oxidative-imbalance markers, we selected 12 two-year-old horses and collected blood samples at various time points before, during (at 5, 10, 15, and 20 km), and after the exercise (at 1, 2, 4, and 6 h post-exercise). These samples were analyzed for haematology, blood biochemistry, antioxidant enzyme activities, and liquid chromatography-mass spectrometry (LC-MS) metabolomics. Our results revealed significant changes in heart rate, speed, blood cells, and biochemical markers throughout the exercise. Antioxidant indicators decreased, while malondialdehyde increased, indicating oxidative imbalance post-exercise. Metabolomics analysis identified 122 differential metabolites, including uric acid and L-tyrosine, which were enriched in pathways related to energy metabolism. Uric acid and tyrosine correlated positively with serum creatine kinase, suggesting their potential as markers of oxidative-imbalance injury. These findings elucidate the mechanisms of endurance adaptability in Mongolian horses and provide a theoretical basis for mitigating oxidative imbalance, enhancing horse performance, and promoting the sustainable development of the equine industry.