Integrated systems pharmacology and transcriptomics to dissect the mechanisms of Loki Zupa decoction in the treatment of murine allergic asthma

Ethnopharmacological relevance: Loki zupa (LKZP) decoction, a traditional Uyghur medicine prescription, has been commonly used to treat numerous respiratory ailments in the Xinjiang region of western China, especially chronic airway inflammatory diseases such as allergic asthma. Due to its complex chemical composition, however, the mechanism of action of LKZP has yet to be fully elucidated. Aim of the study: Based on the balanced regulation theory of pro-inflammation and anti-inflammation, we tried to investigate the effectiveness of LKZP on asthma and its related protective mechanisms. Materials and methods: In this study, an experimental model of asthma was established using ovalbumin (OVA) in BALB/c mice to assess the effects of LKZP. The potential mechanism of LKZP anti allergic asthma were researched by the combination of in silico systems pharmacology and in vivo transcriptomics. Results: Our data revealed that LKZP exerted a therapeutic effect against OVA-induced asthma by reducing airway hyperresponsiveness (AHR), peribronchial inflammation, and mucus hypersecretion. Meanwhile, LKZP downregulated the expression of OVA-induced IgE, interleukin (IL)-4, IL-5, IL-13, and tumor necrosis factor (TNF)-alpha and concurrently promoted the expression of interferon (IFN)-gamma in serum and bronchoalveolar lavage fluid (BALF). Systems pharmacology analysis identified 10 core bioactive ingredients and 26 hub targets of LKZP against asthma. Transcriptomic analysis confirmed 246 differentially expressed genes (DEGs) after LKZP treatment. These were mainly expressed in cytokine-cytokine receptor interactions and immune and inflammatory response-related signaling pathways. Additionally, the real-time quantitative PCR (qPCR) results for the nine selected DEGs matched those of the RNA-seq analysis. Nuclear factor (NF)-kappa B and hypoxia-inducible factor (HIF)1 signaling pathways were identified as candidate targets involved in the action of LKZP on allergic asthma, which was highly consistent with the findings in silico. By qPCR, Western blot, and immunohistochemical analysis, it was verified that LKZP treatment dramatically inhibited the activation of NF-kappa B p65 and HIF-1 alpha stimulated by OVA in asthmatic mice. Conclusions: Taken together, our experimental data revealed that LKZP could be a candidate for the treatment of allergic asthma via NF-kappa B and HIF-1 signaling pathways.

Automated summary

In this study, the therapeutic effect of LKZP on ovalbumin-induced asthma was investigated. LKZP was found to reduce airway hyperresponsiveness, inflammation, and mucus hypersecretion in a mouse model. It also downregulated the expression of IgE, IL-4, IL-5, IL-13, and TNF-alpha, while promoting the expression of IFN-gamma. Systems pharmacology analysis identified key bioactive ingredients and targets of LKZP against asthma. Transcriptomic analysis revealed differentially expressed genes related to cytokine-cytokine receptor interactions and immune and inflammatory response pathways. NF-kappa B and HIF-1 signaling pathways were identified as candidate targets involved in the action of LKZP on allergic asthma. Experimental validation confirmed the inhibition of NF-kappa B and HIF-1 alpha activation by LKZP treatment.