Characterization of chemical components and the potential anti-influenza mechanism of Fructus Arctii by a strategy integrating pharmacological evaluations, chemical profiling, serum pharmacochemistry, and network pharmacology

The presence of chemical components in traditional Chinese medicine (TCM) is the functional basis for its therapy achievement, and the absorbed components under disease conditions mainly contribute to the therapeutic effects. Fructus Arctii (FA) was widely used in traditional Chinese medicine preparation (TCMP) to clear heat or treat influenza. To date, the chemical profile (in vitro and in vivo) and anti-influenza mechanism of FA are still unrevealed, limiting its clinical applications. In this work, an integrated strategy combining pharmacological evaluations, chemical profiling, serum pharmacochemistry, and network pharmacology (PCSN strategy) is proposed and applied to investigate chemical components (in vitro and in vivo), anti-influenza effects, and potential mechanisms of FA. First, the anti-influenza effects of FA were evaluated using an H1N1 virus-infected mouse model. Second, the chemical compounds of FA in vitro and in the disease model were profiled and characterized using liquid chromatography coupled with mass spectrometry. Lastly, the targets and potential mechanisms are predicted based on the absorbed components and a network pharmacology method. As a result, FA (20 g crude drug kg(-1) day(-1)) could significantly improve the survival rate and ameliorate lung inflammation caused by the H1N1 virus. Moreover, a total of 123 compounds (53 potential novel compounds) were identified or tentatively characterized in FA. Furthermore, 124 xenobiotics (38 prototypes and 86 metabolites) were tentatively characterized in the infected mice. The xenobiotics were unitized to predict targets and mechanisms in treating influenza. It was found that FA could target EGFR, CASP3, MAPK1, PTGS2, PIK3CA, ESR1, etc., which are mainly involved in the PI3K-Akt signaling pathway, progesterone-mediated oocyte maturation, etc. Among them, the PI3K-Akt signaling pathway (correlated to PI3K complex) and TNF signalling pathway, are regarded as the core pathways for FA in the treatment of influenza. It is the first time that chemical information is obtained on FA in vitro and in vivo, and the anti-influenza effects and the potential mechanism of FA were also evaluated. Moreover, the potential novel compounds for further validation of FA are also provided.