Based on network pharmacology combined with in vivo and in vitro experiments, this study validated the mechanism of Angelicae Sinensis Radix (AS) in treating knee osteoarthritis (KOA). The active components and targets of AS were retrieved from the TCMSP database. KOA-related gene targets were screened using the TTD, GeneCards, and OMIM databases. Core targets and associated signaling pathways were identified through protein-protein interaction network analysis, along with GO and KEGG enrichment analyses. Molecular docking was then employed to analyze the binding interactions between active components and targets.A KOA rat model was established by sodium iodoacetate (MIA) induction, with knee joint damage assessed via HE staining. Expression changes of core targets were detected using ELISA, Western blot, and RT-qPCR. Rat chondrocytes were isolated, and a KOA chondrocyte model was induced by IL-1β. The optimal intervention concentration of AS was determined using the CCK-8 assay, followed by Western blot analysis of related protein expression.Twenty-six active components of AS and seventy-seven potential therapeutic targets for KOA were identified. These targets were primarily involved in regulating inflammatory responses, apoptotic processes, and the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. Results from both in vivo and in vitro experiments demonstrated that AS: significantly downregulated the expression levels of inflammatory factors; inhibited apoptosis; promoted extracellular matrix anabolism; reduced both the protein expression and phosphorylation levels of PI3K/AKT in chondrocytes; and simultaneously suppressed the mRNA expression of PI3K and AKT. Pathological changes in knee joint tissues and chondrocytes were markedly ameliorated.In conclusion, AS treats KOA through a multi-component, multi-target approach by regulating the PI3K/AKT signaling pathway, achieving multi-dimensional chondroprotective effects.