Jiu-Wei-Yong-An decoction alleviates atopic dermatitis by suppressing TLR4/MyD88/NF-κB signaling pathway

Jiu-Wei-Yong-An (JWYA) decoction is a clinical traditional Chinese medicine prescription for atopic dermatitis (AD). Its anti-inflammatory and anti-pruritic activities have been previously confirmed, yet the underlying molecular mechanism remains unclear.

This work aimed to identify JWYA's bioactive compounds and clarify its therapeutic mechanisms against AD.

We established MC903-induced AD mouse models and TNF-α/IFN-γ-stimulated HaCaT keratinocytes to assess JWYA's efficacy. LC-MS/MS combined with bioinformatics, public microarray datasets, and compound-target prediction was used to screen active components and core pathways. In vivo dermatitis severity and epidermal thickness were quantified after 14-day JWYA oral administration. Western blotting detected pathway protein expression in vitro and in vivo. TLR4 inhibitor TAK-242 rescue assays verified TLR4-dependent regulation. Molecular docking, molecular dynamics (MD) simulations, DARTS and CETSA assays further validated the direct binding of candidate compounds to TLR4/MyD88/NF-κB.

Thirteen absorbable ingredients of JWYA were identified, with the TLR4/MyD88/NF-κB cascade screened as the core anti-AD pathway. JWYA alleviated AD-like lesions, reduced IgE and proinflammatory cytokines, and suppressed TLR4/MyD88/NF-κB overactivation in mice and keratinocytes; TAK-242 assays confirmed JWYA's effects rely on intact TLR4 signaling. Phylliroside and Suspenoidside B exhibited high affinity toward core targets, with direct physical binding validated by MD, DARTS and CETSA.

JWYA ameliorates AD through multi-component, multi-target modulation of the TLR4/MyD88/NF-κB inflammatory axis. Phylliroside and Suspenoidside B are key bioactive substances that directly bind TLR4/MyD88/NF-κB to inhibit NF-κB hyperactivation, relieving AD-related inflammation. This study provides experimental basis for JWYA's clinical application and subsequent monomer drug development.