OBJECTIVE:To establish a method for the simultaneous determination of coumarin compounds in Yifu pill. METH-ODS:HPLC was performed on the column of Inertsil ODS-3 with mobile phase of acetonitrile-water(49∶51,V/V) at a flow rate was 1.0 ml/min,the detection wavelength was 320 nm,column temperature was 30 ℃,injection volume was 20 μl. RESULTS:The linear range was 5.16-206.4μg/ml for osthole (r=0.999 9),37.40-1 495.89μg/ml for isoimperatorin(r=0.999 9) and 9.95-318.34 μg/ml for columbianadin(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recov-eries were 96.05%-103.19%(RSD=2.18%,n=9) for osthole,96.90%-103.09%(RSD=2.07%,n=9) for isoimperatorin, 95.50%-103.57%(RSD=2.15%,n=9)for columbianadin. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and can be used for the simultaneous determination of coumarin compounds in Yifu pill.

ObjectiveTo investigate the effect of Loki Zupa on airway remodeling in asthma based on ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS）combined with network pharmacology and experimental verification. MethodThe chemical constituents in Loki Zupa were identified by UPLC-MS. The potential active constituents of Loki Zupa were screened out based on literature retrieval， oral availability （OB） and drug-likeness （DL） in the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） and Lipinski's rule of five in SwissADEM. The constituent targets of Loki Zupa were obtained through the SwissTargetPrediction. The relevant targets of airway remodeling in asthma were screened out from Online Mendelian Inheritance in Man（OMIM）， GeneCards， DrugBank， and DisGeNET. The STRING was used to conduct protein-protein interaction （PPI） among the main targets. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses were carried out through DAVID. Finally， an asthmatic airway remodeling model was induced by ovalbumin （OVA） in mice， followed by hematoxylin and eosin（HE）， periodic acid Schiff（PAS）， and Masson staining for the observation of the pathological conditions of lung tissues. The inflammatory cells in the bronchoalveolar lavage fluid（BALF） of mice were detected. The protein expression levels in mouse lung tissues were detected by Western blot and key signaling pathways were further determined. ResultEighty-two constituents were detected in the negative ion mode and 74 in the positive ion mode by UPLC-MS. Thirty-six candidate constituents and 578 predicted targets of Loki Zupa were screened out through network pharmacology， and 173 common targets with airway remodeling in asthma were obtained， including key compounds such as sebacic acid， pectolinarigenin， naringenin， apigenin， and potential targets such as protein kinase B1（Akt）1 and hypoxia-inducible factor 1α（HIF-1α）. As predicted by KEGG enrichment analysis， Loki Zupa mainly exerted the effect against airway remodeling in asthma through phosphatidylinositol 3-kinase （PI3K）/Akt， HIF-1α， mitogen-activated protein kinase （MAPK）， and other signaling pathways. Animal experiments showed that the compound formula of Loki Zupa could reduce the proliferation of airway goblet cells in asthmatic mice， improve the deposition of collagen under the airway epithelium， and decrease the up-regulated relative expression levels of phosphorylate（p）-Akt/Akt and HIF-1α by OVA sensitization in mice （P<0.05， P<0.01）， which was consistent with the results of network pharmacology. ConclusionUPLC-MS combined with network pharmacology was used to preliminarily clarify the chemical composition of Loki Zupa and its underlying mechanism in intervention in airway remodeling in asthma. Specifically， Loki Zupa presumably synergistically intervened in airway remodeling in asthma through key targets represented by Akt1 and HIF-1α， and multiple pathways represented by the PI3K/Akt and HIF-lα pathways， which is expected to provide ideas for further research on Loki Zupa.