Integrating Network Pharmacology and MALDI-MSI Spatial Metabolomics Revealing Ellagic Acid as the Key Anti-Pharyngitis Agent in Fructus Chebulae and its Key Metabolic Pathways in Pharyngeal Tissues

Abstract

Background: Current pharyngitis treatment relies heavily on antibiotics and glucocorticoids, leading to increasing bacterial resistance and significant adverse drug reactions. The traditional efficacy of Fructus Chebulae is to reduce swelling and relieve sore throat. Its key components for anti-pharyngitis and mechanism need to be systematically elucidated. Purpose: This study aimed to integrate MALDI-MSI spatial metabolomics and network pharmacology to systematically analyze Fructus Chebulae's therapeutic mechanism against acute pharyngitis, identifying core components of ellagic acid, key targets, and spatial metabolic pathways. Study design: A "disease-component-metabolic pathways" strategy combined network pharmacology screening with in vivo validation in an acute pharyngitis animal model, followed by spatial metabolomics analysis. Methods: Network pharmacology matched Fructus Chebulae's components with disease targets. An acute pharyngitis model was established using 5% ammonia oropharyngeal spray; identified core component of ellagic acid was administered via spraying. Phenotypes were monitored endoscopically. HE staining, serum TNF-α/oxidative stress markers were assessed. MALDI-MSI enabled in-situ metabolite visualization in lesions; differential metabolites were screened with biomarkers identified and metabolic networks constructed. Results: Network pharmacology identified ellagic acid and SRC protein as core ingredient and key target. Ellagic acid spraying dose-dependently repaired pharyngeal/lung damage, significantly reduced TNF-α and oxidative stress, inhibited the expression of SRC protein. MALDI-MSI visualized metabolites, screening 376 differentially expressed metabolites and pinpointing 49 biomarkers. Ellagic acid inhibited inflammation, scavenged radicals, and promoted regeneration by regulating amino acid biosynthesis, riboflavin, and lipid metabolism.