Sangmei Zhike Granules alleviate type 2 inflammation in allergic asthma via the TLR4/PI3Kγ/NF-κB axis: insights from UPLC-Q-Orbitrap-MS, network pharmacology, and experimental validation.

Abstract

BACKGROUND: Allergic asthma is a common allergen-driven phenotype of asthma with a high prevalence. Current first-line therapies are limited by poor treatment adherence, frequent relapse after discontinuation, and heterogeneous therapeutic responses. Sangmei Zhike Granules (SMZK), a traditional Chinese medicine formula formulated to dispel wind, diffuse the Lung, and nourish yin, have demonstrated therapeutic efficacy against allergic asthma; however, their underlying mechanisms remain unclear. OBJECTIVE: This study aimed to identify the lung-distributed constituents of SMZK and elucidate their mechanisms of action against allergic asthma. METHODS: Ultra-performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS) was used to characterize the chemical constituents of SMZK and identify SMZK-derived prototype compounds in rat lung tissue. Network pharmacology analysis was subsequently performed based on the screened prototype compounds to predict potential targets and signaling pathways. An ovalbumin (OVA)-induced rat model of allergic asthma was established, and the therapeutic effects of SMZK were evaluated using pulmonary function testing, histopathological examination, and enzyme-linked immunosorbent assay (ELISA). The predicted signaling pathway was further validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blotting (WB), and ELISA. RESULTS: A total of 812 chemical constituents were identified in SMZK, and 41 SMZK-derived prototype compounds were detected in rat lung tissue. Network pharmacology analysis of 26 prototype compounds that met the screening criteria identified IL6, MAPK8, ESR1, SRC, STAT3, TNF, MAPK3, MAPK1, and RELA as key targets, with the PI3K-Akt and Toll-like receptor signaling pathways emerging as major candidate pathways. SMZK improved pulmonary function, alleviated pulmonary inflammatory cell infiltration, and attenuated type 2 airway inflammation. Mechanistically, SMZK downregulated the expression of TLR4 and PI3Kγ and inhibited NF-κB p65 activation, thereby suppressing the TLR4/PI3Kγ/NF-κB signaling axis and ameliorating type 2 airway inflammation. CONCLUSION: The lung-distributed constituents of SMZK may attenuate type 2 airway inflammation in allergic asthma primarily by suppressing the TLR4/PI3Kγ/NF-κB signaling axis.

Abstract as supplied by the indexing source. © the authors / publisher.