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

Current pharyngitis treatment relies heavily on antibiotics and glucocorticoids, leading to increasing bacterial resistance and significant adverse drug reactions, while the traditional efficacy of Fructus Chebulae is to reduce swelling and relieve sore throat, yet its key components and mechanism against pharyngitis need systematic research. Therefore, this study aimed to integrate MALDI-MSI spatial metabolomics and network pharmacology to systematically analyze Fructus Chebulae's therapeutic mechanism against acute pharyngitis, identifying ellagic acid as the core component, along with key targets and spatial metabolic pathways. Using a ‘disease-component-metabolic pathways’ strategy, network pharmacology was combined with in vivo validation in an acute pharyngitis animal model induced by 5% ammonia oropharyngeal spray, followed by spatial metabolomics analysis. The results showed that network pharmacology identified ellagic acid and SRC protein as the core ingredient and key target. Ellagic acid spraying dose-dependently repaired pharyngeal damage, significantly reduced TNF-α and oxidative stress, and inhibited SRC protein expression. Furthermore, MALDI-MSI visualized metabolites, screening 376 differentially expressed metabolites and pinpointing 49 biomarkers. Specifically, ellagic acid inhibited inflammation, scavenged radicals, and promoted regeneration by regulating amino acid biosynthesis, riboflavin, and lipid metabolism. In conclusion, this study multidimensionally analyzed Fructus Chebulae's traditional efficacy using network pharmacology, confirming the anti-pharyngitis efficacy of ellagic acid, and for the first time via MALDI-MSI illustrated its core spatial metabolic pathways regulating inflammation, oxidative stress, and tissue repair.