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Hence, the mechanism of spleen tissue responding to cold stress in the P. sinensis was investigated. The present results showed that the superoxide dismutase (SOD) activity declined from 4 to 16 days post-cold-stress (dps), while the catalase (CAT) and glutathione peroxidase (GSH-Px) activities increased, from 4 to 8 dps in the 14 \u00b0C (T14) and 7 \u00b0C (T7) stress groups. The spleen transcriptome in the T7 group and the control group (CG) at 4 dps obtained 2625 differentially expressed genes (DEGs), including 1462 upregulated and 1663 downregulated genes. The DEGs were enriched mainly in the pathways \u201cintestinal immune network for IgA production\u201d (Pigr, Il15ra, Tnfrsf17, Aicda, and Cd28), \u201ctoll-like receptor signaling pathway\u201d (Mapk10, Tlr2, Tlr5, Tlr7, and Tlr8), and \u201ccytokine\u2013cytokine receptor interaction\u201d (Cx3cl1, Cx3cr1, Cxcl14, Cxcr3, and Cxcr4). The metabolomic data showed that esculentic acid, tyrosol, diosgenin, heptadecanoic acid, and 7-ketodeoxycholic acid were obviously increased, while baccatin III, taurohyocholate, parthenolide, enterolactone, and tricin were decreased, in the CG vs. T7 comparison. Integrated analysis of the two omics revealed that \u201cglycine, serine and threonine metabolism\u201d, \u201cFoxO signaling pathway\u201d, and \u201cneuroactive ligand\u2013receptor interaction\u201d were the main pathways responding to the cold stress. Overall, this work found that low temperature remarkably influenced the antioxidant enzyme activities, gene expression pattern, and metabolite profile in the spleen, indicating that immunity might be weakened by cold stress in P. sinensis.<\/jats:p>","DOI":"10.3390\/antiox14020217","type":"journal-article","created":{"date-parts":[[2025,2,14]],"date-time":"2025-02-14T06:52:01Z","timestamp":1739515921000},"page":"217","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Molecular Response and Metabolic Reprogramming of the Spleen Coping with Cold Stress in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis)"],"prefix":"10.3390","volume":"14","author":[{"given":"Liqin","family":"Ji","sequence":"first","affiliation":[{"name":"Key Laboratory of Tropical and Subtropical Fishery Resources Application and Cultivation, Ministry of Agriculture and Rural Affairs, Pearl River Fisheries Research Institute, Chinese Academy 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