Explore bioactive ingredients and potential mechanism of Houpo Mahuang decoction for chronic bronchitis based on UHPLC-Q exactive orbitrap HRMS, network pharmacology, and experiment verification

Ethnopharmacological relevance: Chronic bronchitis (CB) affects a growing number of people and may be linked to lung function impairment. The traditional Chinese medicine formula Houpo Mahuang Decoction (HPMHD) has been used for clinical treatment of respiratory diseases for thousands of years. Until now, its bioactive in-gredients, potential targets and molecular mechanism remain unclear.Aim of the study: To investigate the effect of HPMHD on the treatment of CB and explore the bioactive ingredients and possible mechanisms of HPMHD against CB.Materials and methods: UHPLC-Q Exactive Orbitrap HRMS was performed to analyze the chemical components of HPMHD. The mechanism of multiple components, targets and pathways of HPMHD in the treatment of chronic bronchitis were explored by network pharmacology. Additionally, CB mice model induced by lipopolysaccharide (LPS) and smoking was used to evaluate the anti-chronic bronchitis activity of HPMHD in vivo. Pulmonary pa-thology was determined by hematoxylin and eosin (H&E) measurement. The levels of TNF-alpha and IL-6 in lung were measured by ELISA. The immunofluorescence experiments were carried out for the expression of IL-1 beta, TNF-alpha, IL-6 and NF-KB p-P65/P65 in lung. Western blot assays were performed to quantify and visualize the protein expression of NF-KB p-P65/P65 in mice lung.Results: Data showed that 79 compounds were identified in HPMHD. The network pharmacology results showed 53 compounds were hinted their effectivity for the treatment of chronic bronchitis with HPMHD, such as ephedrine, schisantherin A, and honokiol. The main targets were predicted as 37 genes, including TNF, TP53, IL6 and so on. HPMHD ameliorated lung damages in mice and inhibited the NF-KB signaling pathway, one of the pathways plotted by KEGG pathway enrichment analysis, by reducing IL-1 beta, TNF-alpha and IL-6 expression and significantly downregulating the NF-KB p-P65/P65.Conclusion: In summary, the complex chemical components of HPHMD was successfully elucidate by UHPLC-Q Exactive Orbitrap HRMS. The study based on network pharmacology and experiment verification indicated that HPMHD can decreased inflammatory response in lung to treat CB. The underlying mechanism may be related to the reduction of inflammation by down-regulated the NF-KB pathways.