Abstract： Objective： To analyze the action mechanism of Huoxue Huayu Prescription in treating knee osteoarthritis （KOA） based on network pharmacology， molecular docking， and molecular dynamics simulation. Methods： Active components and targets of Huoxue Huayu Prescription were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Universal Protein Database （UniProt）， and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM）. KOArelated targets were obtained from GeneCards， Online Mendelian Inheritance in Man （OMIM）， Therapeutic Target Database（ TTD），and DisGeNET. R 4.4.2 software was used to draw Venn diagrams，and Cytoscape 3.8.2 software was used to construct a "medicine-active component-target-disease" network. Protein-protein interaction （PPI） networks were built using the STRING database. Gene Ontology （GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were performed using R 4.4.2 software. Molecular docking of key targets and active components was conducted using AutoDock Vina software，and molecular dynamics simulations were performed using Gromacs 2022.3 software. Results： A total of 786 active components of Huoxue Huayu Prescription were screened， with 674 intersection targets related to the disease. Major active components such as quercetin， luteolin， and baicalin， as well as core targets including mitogen-activated protein kinase （MAPK） 3， MAPK8， interleukin （IL）-6，and tumor necrosis factor （TNF），were identified. GO enrichment analysis revealed that the key gene functions of Huoxue Huayu Prescription in treating KOA included response to lipopolysaccharide， regulation of apoptotic signaling pathways， and leukocyte migration. KEGG pathway enrichment analysis indicated that the key signaling pathways were TNF，phosphatidylinositol 3-kinase and protein kinase B（ PI3K-Akt），IL-17，and hypoxiainducible factor-1 （HIF-1） signaling pathways. Molecular docking results demonstrated good structure-activity relationships between core targets and active components， and molecular dynamics simulations further confirmed the stable binding of luteolin to TNF. Conclusion：Huoxue Huayu Prescription can act through active components such as quercetin，luteolin，and baicalin，targeting MAPK3，MAPK8，IL-6，and TNF，and regulating the TNF，PI3KAkt， IL-17， and HIF-1 signaling pathways to inhibit inflammatory responses， exert antioxidant effects， and delay the progression of KOA.