Integrating Network Pharmacology, Molecular Docking, Dynamics Simulation and Experimental Validation to Decipher the Antipyretic Mechanisms of Xiaochaihu Granules

Xiaochaihu granules (XCHG), a traditional Chinese herbal formulation, have demonstrated clinical efficacy in fever management, but their precise mode of action remains unclear. Our investigation employed an integrative methodology combining network pharmacology, molecular docking and dynamics simulations), and cellular assays to delineate XCHG's antipyretic mechanisms. Analysis of 18 blood-absorbed components identified from XCHG-treated rat plasma, we identified 17 key targets and 5 key components. GO and KEGG analyses showed that XCHG primarily relates to inflammation, immune regulation, neuroregulation, metabolic control, cell proliferation/apoptosis, and vascular homeostasis, mainly exerting anti-inflammatory effects through these mechanisms. Molecular docking results demonstrated good binding activity between key components and targets, particularly EGFR-Enoxolone, ESR1-Liquiritigenin, and SRC-Baicalein. Subsequent dynamics simulations validated the structural integrity of these ternary complexes. In LPS-stimulated macrophages, XCHG significantly inhibited NO production and downregulation of pro-inflammatory mediators (TNF-α, IL-6, IL-1β, PGE2) and enzymes (iNOS, COX-2). These findings establish that XCHG achieves its antipyretic effects through multi-target engagement of key components with key proteins, subsequently modulating inflammatory cascades and cytokine networks, thereby providing mechanistic support for its clinical application in fever treatment.