基于UFLC-MS/MS及生物信息学探究黄龙止咳口服液治疗咳嗽变异性哮喘的药效物质基础及作用机制OA

Aim To explore the pharmacodynamic material basis and mechanism of action of Huanglong cough oral liquid(HL)in treating cough variant asthma(CVA).Methods CVA rats model was estab-lished using ovalbumin(OVA)sensitization.Ultra-fast 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.Bioinformat-ics was employed to screen for potential therapeutic tar-gets of HL.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in lung tis-sue.Capsaicin and acetylcholine challenges were ap-plied to observe changes in cough frequency and the airway hyperresponsiveness(AHR)indicator en-hanced 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.Results The serum concentra-tions of the 9 HL components showed an increasing trend over time,reaching a steady-state concentration by d 7 of administration.HL dose-dependently re-versed 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 identi-fied six highly relevant potential targets:transient re-ceptor potential ankyrin 1(TRPA1),transient recep-tor 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(PT-GIR).HL and its nine components dose-dependently regulated the gene and protein expression of five poten-tial 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.Conclusions 9 components including ephedrine are likely the pharma-codynamic 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.