The pharmacodynamic material basis and mechanism of Huanglong cough oral liquid in treating cough variant asthma via UFLC-MS/MS and bioinformatics

AimTo explore the pharmacodynamic material basis and mechanism of action of Huanglong cough oral liquid (HL) in treating cough variant asthma (CVA).MethodsCVA rats model was established using ovalbumin (OVA) sensitization. Ultrafast liquid chromatography tandem mass spectrometry (UFLC-MS/MS) and enzyme-linked immunosorbent assay (Elisa) were used to detect changes in serum levels of ephedrine and eight other components, as well as interferon-γ (IFN-γ), interleukin-4 (IL-4), and tumor necrosis factor-α (TNF-α) on day 1, 7, and 14 after intragastric administration of HL. Bioinformatics was employed to screen for potential therapeutic targets of HL. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in lung tissue. Capsaicin and acetylcholine challenges were applied to observe changes in cough frequency and the airway hyperresponsiveness (AHR) indicator enhanced pause (Penh) before and after HL and active component intervention. RT-qPCR and Western blot were further used to detect the expression of target genes and proteins.ResultsThe serum concentrations of the 9 HL components showed an increasing trend over time, reaching a steady-state concentration by d 7 of administration. HL dose-dependently reversed the abnormal changes in serum IFN-γ, IL-4, and TNF-α levels in CVA rats, with the middle-and high-HL groups approximating those of the normal group by day 7 and 14. Bioinformatics analysis identified six highly relevant potential targets: transient receptor potential ankyrin 1 (TRPA1), transient receptor potential vanilloid 1 (TRPV1), Erb-B2 Receptor Tyrosine Kinase 2 (ERBB2), Nitric Oxide Synthase 2 (NOS2), cytoplasmic phospholipase A2-IVA (PLA2G4A), and Prostaglandin I2 Receptor (PTGIR). HL and its nine components dose-dependently regulated the gene and protein expression of five potential targets (excluding TRPV1), thereby reducing cough frequency and inhibiting AHR to alleviate CVA. The therapeutic effect of HL was superior to that of the individual nine components.Conclusions9 components including ephedrine are likely the pharmacodynamic material basis of HL for treating CVA. HL exhibits a non-linear concentration-effect relationship, characteristic of multi-component herbal therapeutics. The therapeutic effect of HL is closely related to the regulation of immune-inflammatory responses.