This study comprehensively applied a diverse array of widely targeted metabolomics, network pharmacology, and molecular docking techniques to delve into the potential anti-fatigue mechanism of Crassostrea gigas. The aim was to provide a robust theoretical and experimental foundation for the scientific utilization of Crassostrea gigass. Utilizing an ultra-high performance liquid chromatography-electrospray ionization-triple quadrupole/linear ion trap mass spectrometry (UPLC-ESI-QTRAP-MS/MS) system and database, an extensive targeted metabolomics analysis of the simulated digestive products of Crassostrea gigas in vitro was conducted. This analysis aimed to obtain component transformation information during Crassostrea gigas digestion. Network pharmacological analysis of the anti-fatigue properties of Crassostrea gigas active ingredients was carried out using PubChem, Swiss Target Prediction, GeneCards, and Cytoscape software. From this, a network map of "active ingredients-target" for Crassostrea gigas was constructed. Subsequently, the core targets was verified using molecular docking technology. 1231 active ingredients in 17 categories of Crassostrea gigas were identified. Among these, 106 intersection targets with potential anti-fatigue effects were pinpointed, involving 181 key active ingredients, primarily including 1,5-pentenediamine, tryptamine, 4-tert-octylphenol, etc. Core targets included Estrogen receptor 1 (ESR1), epidermal growth factor receptor (EGFR), and peroxisome proliferator-activated receptor gamma (PPARG). Signal pathway enrichment analysis revealed that the calcium signaling pathway, neuroactive ligand-receptor interaction, and PI3K-Akt signaling pathway were pivotal signaling pathways for fatigue resistance in Crassostrea gigas. Molecular docking results further verified the good combination of key active ingredients and core targets of oyster. Crassostrea gigass may exert an anti-fatigue effect through a synergistic action involving multiple components, targets, and pathways. This study offers theoretical support for further exploration of the active components and molecular mechanisms of Crassostrea gigas, paving a solid foundation for subsequent research and application.