Integrating Serum Pharmacochemistry, Network Pharmacology, and Metabolomics to Explore the Therapeutic Mechanism of Ardisia crenata Sims in Acute Pharyngitis via Regulation of the PI3K/Akt Pathway

Background/Objectives: Ardisia crenata Sims (ACS) is widely used in China for treating sore throat and pharyngeal swelling, but the bioactive constituents and mechanisms underlying its efficacy against acute pharyngitis remain unclear. This study combined UPLC-Q-TOF-MS analysis with integrated network pharmacology, metabolomics, and animal experiments to systematically elucidate the material basis and mechanism of ACS in treating acute pharyngitis. Methods: An integrative strategy combining UPLC-Q-TOF-MS-based serum pharmacochemistry, network pharmacology, metabolomics, and in vivo validation was employed. Acute pharyngitis was induced in rats. Therapeutic effects were evaluated by serum inflammatory cytokine levels and histopathological examination. Chemical constituents and absorbed blood components were identified via UPLC-Q-TOF-MS. Putative targets were predicted through network pharmacology, while metabolomics identified differential metabolites and disturbed pathways. Molecular docking was used to assess binding affinities between core compounds and key targets. Immunohistochemistry was used to assess PI3K-Akt pathway-related protein expression in pharyngeal tissue. Results: A total of 73 chemical constituents were identified from ACS, with 14 prototype compounds confirmed to be absorbed into the bloodstream. ACS significantly regulated serum TNF-α, IL-1β, and IL-6, reduced inflammatory cell infiltration, and alleviated pharyngeal mucosal damage. Integrated analysis revealed 19 differential metabolites and 10 hub genes (AKT1, PIK3CA, EGFR, SRC, etc.), primarily enriched in the PI3K-Akt signaling pathway, purine metabolism, and amino acid metabolism. Molecular docking showed strong binding affinities of quercetin, myricetin, and naringenin with key targets. Immunohistochemistry confirmed that ACS significantly downregulated PI3K, p-PI3K, AKT, and p-AKT expression at the pathway level. Conclusions: ACS exerts anti-pharyngitis effects through its flavonoid-rich bioactive constituents, which show high-affinity binding to the upstream kinases EGFR and SRC, thereby indirectly suppressing the overactivation of the PI3K-Akt signaling pathway and modulating inflammation-related metabolic networks, including endohydrolysis of 1,4-alpha-D-glucosidic linkage, purine metabolism, and glycine, serine, alanine, and threonine metabolism.