Phytomedicine : international journal of phytotherapy and phytopharmacology · 2026
Multi-omics reveals Huanglian ointment supercritical extract alleviates allergic rhinitis involving MAPK/RelB-NF-κB pathway.
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Bai Xueyuan, Shang Liying, Zhang Lequn, Wang Xin, Xu Yanan, Zhao Daqing, Zhang He, Zhang Wei
Аннотация
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.
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.