Abstract：Objective：To analyze the mechanism of Sishen Decoction in the treatment of knee osteoarthritis（ KOA） based on network pharmacology and experimental validation. Methods：The active components of Sishen Decoction and their corresponding targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. Standardized gene names for the targets were obtained from the UniProt database. Disease targets for KOA were collected from the GeneCards database，and overlapping targets between the drug and the disease were identified. The "drug-active components-targets-disease" network was constructed using Cytoscape software. A protein-protein interaction （PPI） network was built using the STRING database to identify core targets. Gene Ontology （GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were performed using the Bioconductor package in R software. Molecular docking was employed to verify the binding affinity between core components and key targets. Knee articular chondrocytes from neonatal rats were extracted， cultured， and stimulated with lipopolysaccharide （LPS） to establish an in vitro model. A drug-containing serum of Sishen Decoction was prepared and used for intervention to experimentally validate the predicted mechanisms. Results： Sixty-three active components and 205 corresponding targets of Sishen Decoction were screened. A total of 3 572 KOA-related targets were obtained. After intersection，130 overlapping targets were identified. The main active component were quercetin，kaempferol and luteolin，etc. PPI analysis indicated that the core targets of Sishen Decoction for KOA included the inflammatory factors interleukin （IL）-6，IL-1β，and tumor necrosis factor -α （TNF-α），etc. GO enrichment analysis showed these targets were significantly enriched in functions closely related to inflammation and metabolic regulation， such as DNA-binding transcription factor binding， ubiquitin-like protein ligase binding， cytokine-cytokine receptor interaction，and kinase regulator activity. KEGG enrichment analysis revealed associations with signaling pathways including IL-17， TNF， Toll-like receptor， T helper （Th）17 cell differentiation， phosphatidylinositol 3-kinase （PI3K）-protein kinase B （Akt）， T cell receptor， mitogen-activated protein kinase （MAPK）， and nuclear factor （NF）-κB. Molecular docking results showed the binding energies between core active components and key targets were all less than -5 kcal/mol. Experimental validation results demonstrated that LPS stimulation significantly increased the expression of inflammatory factors IL-6，IL-1β，and TNF-α in chondrocytes， and intervention with Sishen Decoction drug-containing serum effectively inhibited the release of these inflammatory factors. Conclusion：Based on network pharmacology analysis and experimental validation，the mechanism of Sishen Decoction in the treatment of KOA involves inhibiting the secretion of inflammatory factors and the abnormal activation of inflammatory signaling pathways.