Guominkang formula alleviates airway inflammation in HDM-induced asthma mice by regulating Wnt/β-Catenin pathway.

backgroundThe Guominkang formula (GMK), formulated according to the principle of "treatment based on constitution differentiation," comprises Prunus mume (Siebold) Siebold & Zucc. (Wumei), Saposhnikovia divaricata (Turcz. ex Ledeb.) Schischk. (Fangfeng), Ganoderma lucidum (Curtis) P. Karst. (Lingzhi), and Periostracum Cicadae (Chantui). Clinically, GMK has been shown to modulate allergic constitution, effectively treating allergic asthma (AA) and various other allergic conditions, with a favorable safety profile and substantial therapeutic benefits. However, the precise mechanisms underlying its immune-modulatory effects, particularly in the context of AA, remain inadequately defined.

aimsThis study aimed to investigate the therapeutic effects and underlying mechanisms of GMK in a mouse model of AA.

methodsThe components of GMK were analyzed via LC-MS/MS. AA was induced in female mice through nasal instillation of house dust mites (HDM). Therapeutic efficacy was assessed through histopathological examination of lung tissue, measurement of airway hyperresponsiveness (AHR), and analysis of inflammatory cell infiltration, including eosinophils, neutrophils, macrophages, and subsets of T cells (Th1, Th2, Th17, and Treg). Serum levels of total IgE, HDM-specific IgE (HDM-sIgE), and cytokines (IL-1β, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17, and IFN-γ) were quantified. Additionally, gut microbiome composition and differences between experimental groups were analyzed. Lung tissue transcriptomics identified differentially expressed genes (DEGs) and related signaling pathways. Western blot analysis was performed to evaluate protein expression levels of the Wnt/β-Catenin signaling pathways, contributing to the understanding of GMK's anti-asthma effects. Molecular docking studies were conducted to explore the binding interactions between GMK and the Wnt3a protein.

resultsFourteen compounds were identified in GMK. The formula exhibited significant therapeutic effects in an AA mouse model, evidenced by a reduction in Th2 and Th17 cell populations, restoration of the Th1/Th2 and Th17/Treg immune balance, alleviation of eosinophilic airway inflammation, and a decrease in total IgE and HDM-sIgE levels in serum. GMK also downregulated the expression of IL-1β, IL-4, IL-5, IL-6, IL-10, IL-13, and IL-17, while upregulating IFN-γ expression. Among the various doses, the medium dose proved most effective in mitigating airway inflammation, reducing airway remodeling, and decreasing AHR. Microbiome analysis revealed that GMK treatment reversed the reduced abundance of Firmicutes and Dubosiella in asthma mice, while increasing the abundance of Bacteroidetes and Norank_f_Muribaculaceae. Transcriptomic analysis demonstrated that, compared to asthma mice, DEGs in the lung tissue of GMK-treated mice were primarily enriched in the Wnt and related pathways. Furthermore, GMK modulated the Wnt/β-catenin signaling pathway to treat AA. Molecular docking studies confirmed predicted strong binding interactions between multiple bioactive compounds in GMK and the Wnt3a protein.

conclusionsGMK regulates Th immune balance by modulating the Wnt/β-catenin signaling pathway, thereby reducing airway inflammation in HDM-induced asthma mice. Targeting the Wnt/β-catenin signaling pathway in the lungs may offer a novel therapeutic approach for allergic asthma treatment.