Objective To explore the mechanism of action of astaxanthin (AST) in alleviating renal ischemia-reperfusion injury (RIRI) through an integrated approach of network pharmacology, molecular docking, and experimental validation. Methods Potential targets of AST and RIRI were identified using databases, and the intersection of these targets was used to construct a protein-protein interaction (PPI) network. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed, and molecular docking was conducted between AST and key targets. Forty male SD rats were randomly divided into four groups (n=10 per group): Control, Model, AST, and Model+AST. The Model and Model+AST groups were subjected to exhaustive exercise to induce RIRI. The AST and Model+AST groups received AST at a dose of 25 mg/kg body weight via gavage daily for 4 weeks, while the Control and Model groups received an equivalent volume of olive oil. After 4 weeks, kidney tissues and serum samples were collected from the rats. Histopathological changes in kidney tissues, levels of oxidative stress and inflammation-related factors were assessed using HE staining, ELISA, RT-qPCR, and Western blot. Results (1) Network pharmacology identified 88 potential targets for AST in improving RIRI, with AKT1, NF-κB, CXCL8, and TLR4 emerging as key targets. GO and KEGG analyses indicated that these targets are mainly associated with oxidative stress, inflammatory response, and signaling pathways such as PI3K-AKT, Toll-like receptor, and NF-κB. (2) Molecular docking results showed good binding affinity between AST and the top ten core targets. (3) Experimental results revealed that compared to the Control group, the Model group exhibited significant renal pathological damage and changes in oxidative stress and inflammation-related factors, with upregulated expression of proteins in the TLR4/TRAF6/NF-κB pathway. Compared to the Model group, these indicators were significantly reversed in the Model+AST group. Conclusion AST may improve RIRI by reducing oxidative stress and inflammatory responses through the TLR4/TRAF6/NF-κB pathway, effectively improving renal tissue pathological damage in RIRI rats.