Zhuangdu Quhan Mixture Alleviates Ankylosing Spondylitis in Mice by Inhibiting the Interleukin-17 Signaling.

Abstract

This study aimed to elucidate the therapeutic effect and mechanism of Zhuangdu Quhan Mixture (ZDQHM) in ankylosing spondylitis (AS). An AS mouse model was established in mice using β-glucan. Therapeutic efficacy was assessed through arthritis scores, histopathology, serum cytokine measurement by ELISA, and bone metabolism-related protein analysis via Western blot. Active compounds were identified by UHPLC-MS, with targets predicted using network pharmacology and validated by molecular docking and molecular dynamics simulations. ZDQHM significantly ameliorated spinal inflammation and histopathological damage, reduced serum IL-6 and TNF-α levels, and modulated bone metabolism proteins. Network pharmacology identified 45 intersecting targets between ZDQHM compounds and AS, predominantly enriched in IL-17, Toll-like receptor, and NOD-like receptor pathways. Molecular docking and dynamics demonstrated binding of key compounds to IL-17A. In vivo validation revealed that ZDQHM suppressed IL-17A, Act1, and IL-17RA expression, and recombinant IL-17A partially reversed its therapeutic effects. ZDQHM alleviates inflammation and abnormal bone metabolism in AS mice, potentially through inhibition of the IL-17 signaling pathway. Beyond confirming the established role of IL-17 in AS, this study provides an integrated pharmacological framework explaining how ZDQHM exerts its effects via regulation of the IL-17A/IL-17RA/Act1 axis, offering mechanistic insights and experimental support for TCM in AS therapy.