基于UPLC-Q-Orbitrap HRMS、网络药理学与SPR的小儿止哮平喘颗粒抗哮喘机制研究OA

[Objective]To analyze the material basis and mechanism of Xiao'er Zhixiao Pingchuan Granules on bronchial asthma(BA)through ultra-performance liquid chromatography-quadrupole orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap HRMS),network pharmacology,molecular docking and surface plasmon resonance(SPR)technology,combined with animal experiments.[Methods]First,UPLC-Q-Orbitrap HRMS method was used to detect the chemical components and migrating components of Xiao'er Zhixiao Pingchuan Granules;secondly,using network pharmacology to predict potential targets and pathways for the treatment of BA by migrating components;Then,the binding activity between the active ingredients and the core targets was validated using molecular docking and SPR techniques;Finally,the expression of key target proteins was confirmed through an asthma mouse model and western blot analysis.[Results]A total of 112 compounds and 79 serum migrating components were identified in Xiao'er Zhixiao Pingchuan Granules.Network pharmacology analysis indicated that wogonin and tanshinone ⅡA were the core active ingredients,and their effects might be associated with signaling pathways such as phosphoinositide 3-kinase/protein kinase B(PI3K-Akt),mitogen-activated protein kinase(MAPK),and transient receptor potential(TRP).Molecular docking and SPR assays confirmed that tanshinone ⅡA exhibited moderate binding affinity to nerve growth factor(NGF).Animal experimental results showed that Xiao'er Zhixiao Pingchuan Granules significantly inhibited the protein expression of NGF,tropomyosin receptor kinase A(TrkA),transient receptor potential vanilloid 1(TRPV1),substance P(SP),and mas-related G protein-coupled receptor X2(MRGPRX2)in the lung tissue and dorsal root ganglia of asthmatic mice,and also suppressed the phosphorylation of p38 mitogen-activated protein kinase(p38 MAPK).[Conclusion]Xiao'er Zhixiao Pingchuan Granules may alleviate asthmatic symptoms by regulating the NGF/TrkA/TRPV1/SP/MRGPRX2 signaling axis and ameliorating neuroimmune-derived inflammation.