Intensive aquaculture is frequently challenged by hypoxia, which has emerged as a critical environmental issue. The pikeperch (Sander lucioperca), a globally recognized high-value aquaculture species, exhibits high sensitivity to hypoxia. However, the physiological mechanisms of pikeperch under hypoxia and reoxygenation stress remain unclear. This study investigated the effects of hypoxia exposure (6, 24, and 48 h), reoxygenation (6 h after 48 h hypoxia), and normoxia on liver histology, biochemical parameters, and transcriptome in pikeperch. Results showed that hypoxic stress markedly elevated hepatic SOD, CAT, and GSH-Px activities in pikeperch. Prolonged hypoxic stress elevated ALT, AST, and MDA levels, indicating gradual liver injury, which was partially alleviated after reoxygenation. Transcriptome sequencing of pikeperch liver identified 6897 differentially expressed genes, as well as two gene expression profiles and two hypoxia-reoxygenation functional modules. Transcript expression analysis revealed time-dependent metabolic gene responses under stress: hypoxia upregulated glycolysis-related genes (HK, PFKP, LDHA) at 6 h, while genes involved in fatty acid β-oxidation (ACADS, ACADM, CPT1A, CPT2) were induced at 24 h and further upregulated following reoxygenation. Several immune-related genes exhibited decreased expression under hypoxia, including TLR3, IL1R1, IL17RC, NLRP3, NLRP6, and NLRP12. HIF-1α pathway genes displayed distinct temporal expression patterns: genes related to the HIF-1α pathway (HIF-1α, PHD2, PHD3, VHL) showed an early response at 6 h of hypoxia. PHD2/3 were significantly downregulated at 48 h, and their expression was not fully restored upon reoxygenation. These results suggest that hypoxia may trigger a potential tendency toward sequential metabolic reprogramming from glycolysis to fatty acid β-oxidation at the transcriptional level, accompanied by oxidative stress, liver injury, and altered immune-related signaling, which are partially ameliorated after reoxygenation. The present work offers valuable insights into the molecular regulatory mechanisms of acute hypoxia-reoxygenation stress in pikeperch.