Exploring the Therapeutic Potential and Underlying Mechanism ofCallerya speciosa in Ankylosing Spondylitis: Network Pharmacology,Molecular Docking, and Single-Cell Sequencing

Introduction: Callerya speciosa (C. speciosa), a traditional herbal medicine, is widely used in the treatment of Ankylosing Spondylitis (AS); however, its active components and therapeutic mechanisms remain unclear.

Methods: The present work collected active compounds of C. speciosa, potential drug targets, and AS-related genes from literature and databases. Obtained Differentially Expressed Genes (DEGs) in AS through single-cell sequencing analysis, and identified intersecting targets among compounds, diseases, and DEGs. Protein-Protein Interaction (PPI) networks were constructed using STRING and Cytoscape to identify hub genes. Functional enrichment analysis (GO/KEGG) was performed via Metascape. Summary-data-based Mendelian Randomization (SMR) was applied to identify core targets with causal associations with AS. Molecular docking validated compoundtarget interactions.

Results: Formononetin was identified as the key active compound. PPI and SMR analyses revealed TLR2, JAK2, and IL7R as core targets, whose elevated expression correlated with an increased risk of AS. GO/KEGG analysis indicated multi-target modulation of pathways, including Toll-like receptor signaling and JAK-STAT cascades. Molecular docking confirmed strong binding between Formononetin and these targets.

Discussion: By integrating network pharmacology, SMR analysis, and single-cell sequencing studies, further insights were gained into how C. speciosa intervenes in the pathological processes associated with AS through a multi-component, multi-target, and multi-pathway collaborative action. These findings provide crucial theoretical and scientific support for the potential use of C. speciosa as a therapeutic agent for AS.

Conclusion: This study elucidates the multi-component, multi-target mechanism of C. speciosa against AS, highlighting Formononetin's role in regulating TLR2, JAK2, and IL7R. These findings provide a scientific foundation for developing novel AS therapies based on C. speciosa.