Objective  Combining network pharmacology and Mendelian randomization (MR) approaches to systematically elucidate the potential mechanisms of Shaoyao decoction (SYD) in the treatment of knee osteoarthritis (KOA).

Methods  Active compounds of SYD and their putative targets were retrieved from the TCMSP database and intersected with KOA-related targets to identify shared drug–disease targets. These targets were imported into the STRING database and Cytoscape to construct a protein-protein interaction (PPI) network, and key regulatory targets were identified using topological analysis. GO enrichment and KEGG pathway analyses were subsequently performed. Furthermore, MR analysis was conducted using genome-wide association study data, with single nucleotide polymorphisms (SNPs) as instrumental variables, to evaluate the causal relationship between core targets and KOA.

Results  A total of 103 active compounds and 233 potential targets of SYD were identified, of which 133 overlapped with KOA-related targets. Quercetin was identified as the primary active compound, while F7, PTGS2, ESR1, HSP90AA1, and SCN5A were recognized as core targets. PPI network analysis further highlighted IL-6, TNF, AKT1, IL-1β, and MMP-9 as potential key targets. GO enrichment yielded 2,632 entries, including 2,417 biological processes, 147 molecular functions, and 68 cellular components. The KEGG enrichment results showed that 183 signaling pathways were involved, including atherosclerosis, TNF signaling pathway, HIF- 1 signaling pathway, IL-17 signaling pathway, etc. MR analysis identified 26 SNPs significantly associated with KOA (all P<5×10- 5), among which SCN5A, F7, and HSP90AA1 showed positive causal associations with KOA risk.

Conclusion  This study, through an integrated analysis of network pharmacology and MR, further reveals that key targets such as HSP90AA1, F7, and SCN5A exhibited genetic causal associations with the risk of KOA, providing a theoretical reference for the pharmacological mechanisms of Shaoyao Decoction.

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