Integrating transcriptomics and metabolomics to explore the therapeutic effects and mechanisms of Zhushagen-Shandougen herb pair on chronic pharyngitis.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Ardisia crenata Sims (Zhushagen) and Sophora tonkinensis Gagnep. (Shandougen), core components of Kaihoujian Spray, are used in Miao medicine to treat chronic pharyngitis (CP), but their mechanisms remain unclear. AIM OF THE STUDY: This study aimed to investigate the therapeutic effects of Zhushagen, Shandougen, and their combination (Zhushagen-Shandougen) on CP, and to elucidate the pharmacological mechanisms via serum metabolomics and pharyngeal mucosa transcriptomics. METHODS: A rat model of chronic pharyngitis was induced by ammonium hydroxide administration. Therapeutic effects were evaluated via behavioral assessment, histopathological examination, and ELISA for inflammatory cytokines. For mechanism exploration, transcriptomic analysis of pharyngeal mucosa and metabolomic analysis of serum were performed in rats. Correlation analysis was used to integrate transcriptomic and metabolomic data, and a core regulatory network was constructed based on key genes, metabolites, and pathways. Potential targets were validated by immunohistochemistry and RT-qPCR. RESULTS: All treatments dose-dependently ameliorated CP symptoms, reduced the pro-inflammatory cytokines IL-1β, IL-6, PGE2, and TNF-α, increased IL-10, and alleviated mucosal lesions, with the combination being the most potent. In omics analyses, 32 differential metabolites and 121 differential mRNAs were identified. KEGG enrichment showed that differential metabolites were primarily involved in riboflavin and glutathione metabolism, while differential mRNAs were enriched in the MAPK and PI3K-Akt signaling pathways. RT-qPCR validation of 9 genes confirmed the transcriptomic results; notably, the mRNA expressions of p38, erk1/2, pi3k, and akt were altered. Immunohistochemistry further confirmed changes in the protein levels of p38, ERK1/2, and Akt, as well as the activations of p38 and PI3K, validating the omics findings. CONCLUSION: Zhushagen, Shandougen, and Zhushagen-Shandougen exert therapeutic effects on CP. The present study found that Zhushagen-Shandougen may suppress the PI3K/Akt and MAPK signaling pathways, thus alleviating ammonium hydroxide-induced CP. The elucidated mechanism provides a significant theoretical foundation and a novel therapeutic approach for the treatment of CP.