Multi-omics reveals Huanglian ointment supercritical extract alleviates allergic rhinitis involving MAPK/RelB-NF-κB pathway.

BACKGROUND

Huanglian ointment, recorded in the 'Medical Canon of the Golden Mirror', is traditionally used to treat nasal sores and damp-heat conditions. Allergic rhinitis (AR) is a common immune-mediated disease. However, the mechanism by which the supercritical extract of Huanglian ointment (HLGSE) relieves AR remains unclear.

PURPOSE: This study aimed to investigate the effects of HLGSE on AR and its underlying mechanisms.

METHODS

Huanglian ointment was extracted by various methods, and the resulting extracts were screened for activity in HMC-1 and RAW264.7 inflammatory models. Optimal extraction conditions were identified through orthogonal design, and extract composition was analyzed by GC-MS, GC-IMS, and E-nose. HLGSE's anti-inflammatory efficacy was then validated in vivo and in vitro.

RESULTS

HLGSE was the most effective extract, rich in esters, ketones, and alcohols. In vitro and in vivo, HLGSE reduced inflammation and alleviated AR in BALB/c mice. We combined network pharmacology, 16S rRNA gene sequencing, four-dimensional data-independent acquisition (4D-DIA) proteomics, and metabolomics. 16S rRNA sequencing identified Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria as the predominant phyla; proteomics revealed key signaling pathways, including cGMP-PKG, cAMP, MAPK, NF-κB, and PI3K; and metabolomics characterized the major metabolic pathways. HLGSE modulated MAPK and NF-κB signaling pathway. Gain- and loss-of-function experiments further indicated that RelB modulation altered NF-κB-associated inflammatory responses in LPS- and IFN-γ-stimulated HMC-1 cells.

CONCLUSION

This is the first study to combine GC-MS, GC-IMS, and E-nose to characterize HLGSE's components. HLGSE can remodel the nasal microbiota, serum metabolites, and nasal mucosal proteins and suppress the MAPK and NF-κB signaling pathways, potentially alleviating AR.