Non-targeted LC-MS Metabolomics Reveals Serum Metabolites for High-altitude Adaptation in Tibetan Donkeys

Tibetan donkeys inhabit the harsh environment of the Qinghai-Tibet Plateau. Research on serum metabolites related to their high-altitude adaptation is limited compared to other livestock. We used liquid chromatography-mass spectrometry (LC-MS) to analyze serum samples from healthy adult donkeys in Shigatse, Changdu, and Dezhou to evaluate the effects of high altitudes on serum metabolites. Metabolomics analysis identified 443 differential metabolites (DMs) across the three groups, meeting criteria of VIP ≥ 1, p-value < 0.05, and Fold-Change ≥ 1.2 or ≤ 0.5. Significant upregulation was observed in deoxycholic acid, vitamin A, vitamin C, flavin mononucleotide, n-acetylserotonin, N'-formyl-kynurenine, calcidiol, and adenosine monophosphate in the high-altitude group compared to the low-altitude control group. The DMs were involved in processes such as bile secretion, vitamin digestion and absorption, tryptophan metabolism, and parathyroid hormone synthesis, secretion, and action. All these are crucial for nutrient metabolism, immune function, and antioxidant stress response. This study constructed a metabolomics dataset of Tibetan donkey serum and revealed differential metabolites among donkeys from different geographic regions and environments. The results offer crucial insights into the adaptive regulatory mechanisms.