| Objective: This study established a method based on metabolomics and network pharmacology to analyze the endogenous metabolites of A549 cells after intervention with usnic acid and to explore the therapeutic mechanism of usnic acid on Non-Small Cell Lung Cancer.Methods: Firstly,lung cancer A549 cells were treated with different concentrations of usnic acid to evaluate its effect.The determined 80% survival rate value was used for the cells in the treatment group,and the cells in the control group were normal A549.Then,after 24 h of intervention with this concentration of usnic acid,endogenous substances were obtained.The samples were analyzed with UHPLC-Q-TOF-MS/MS platform.The data were detected by the tool with software Progenesis QI.Statistical analysis t-test and OPLS-DA method were used to obtain P and VIP value with SIMCA-P14.1 software.Metabolites that satisfy both P <0.05 and VIP> 1 were considered differential metabolites.Potential biomarkers were obtained after identified through the HMDB database.Finally,the pathway enrichment analysis and screening were performed through the online platform Metabo Analyst 4.0 and the KEGG online database.In the research of network pharmacology,the target genes of usnic acid and NSCLC were identified by some online databases,and then the core target genes were screened by Cytoscape 3.7.1 software.Then,the Protein-Protein-Interaction network was constructed.The pathways were analyzed using the Metascape database for further study.At last,network models of usnic acid in treatment of NSCLC were constructed.And related biological processes that may be affected were also predicted.Results: The concentration of usnic acid that made A549 cells reached 80% survival rate was 50 ?g / m L.With this concentration as treatment group,47 potential biomarkers were screened and identified through multivariate statistical analysis and HMDB database.The metabolic pathways involved energy metabolism,amino acid metabolism and lipid metabolism.With network pharmacology,core target genes were screened for usnic acid and NSCLC,involving 20 major related pathways.Metabonomics and network pharmacology have focused on the mechanisms of action and complement each other,showing possible mechanisms at the molecular and pathway levels of usnic acid in treating NSCLC.Conclusions: In this study,a comprehensive method based on metabolomics and network pharmacology analysis was established for the first time,using A549 cell model to explore the therapeutic mechanism of usnic acid in NSCLC.As a result,47 differential metabolites were screened which were involved glutathione metabolism,amino acid metabolism,glycerophospholipid metabolism and other metabolic pathways.Therefore,the combination of metabolomics and network pharmacology provided a sufficient explanation in the therapeutic mechanism of usnic acid in NSCLC.And this method also provided a new clue for the research and explanation of drug action mechanism.. |
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