Exploration of Active Components in Hou-Yan-Qing Oral Liquid and Its Alleviative Effect on Acute Pharyngitis via NRF2/NF-κB Pathways.

Abstract

The Hou-Yan-Qing oral liquid (HYQ) is a popular traditional Chinese medicine for the treatment of pharyngitis. However, due to its complex composition, little attention has been directed toward the analysis of its mechanism and active components. An ammonia-induced acute pharyngitis rat model was established, with HYQ's effects and mechanism assessed via histopathology, enzyme-linked immunosorbent assay, and Western blot. Furthermore, the active components of HYQ were analyzed using HPLC and ultra performance LC (UPLC)-QTOF-MS in conjunction with an in vitro lipopolysaccharide-induced RAW264.7 cell model. Treatment with HYQ significantly reduced the levels of cytokines such as pro-inflammatory cytokines (interleukin-6 [IL-6], tumor necrosis factor α [TNF-α]) and inflammatory mediators (Prostaglandin E2 [PGE2]), inhibited the expression of cyclooxygenase-2 (COX-2), 5-LOX, p-IKKβ, p-nuclear factor-kappaB (NF-κB) and p-inhibitor of kappaBα (p-IκBα) proteins, and promoted the expression of nuclear factor-E2-related factor 2, heme oxygenase-1, and NAD(P)H:quinone oxidoreductase 1 proteins. Chemical profiling of ten batch HYQ specimens via HPLC and UPLC-QTOF-MS established characteristic fingerprints and identified 24 shared components. Using spectrum-effect relationship modeling, which employed gray relational analysis to correlate chromatographic peaks with bioactivity indices, we found that components such as 25S-Inokosterone and β-ecdysterone may be the key active components of HYQ's anti-inflammatory effect. Subsequent validation confirmed that β-ecdysterone (1-100 μM) exhibited no cytotoxicity in RAW264.7 cells. Furthermore, treatment with β-ecdysterone significantly reduced levels of pro-inflammatory cytokines (IL-6, TNF-α) and inflammatory mediators (PGE2), and inhibited the expression of COX-2, 5-LOX, p-IKKβ, p-NF-κB, and p-IκBα proteins.