Combining LC/IM-QTOF-MS with serum pharmacochemistry to identify active compounds and functional mechanisms of...

Fritillaria ussuriensis Maxim. (FUM) is a traditional medicinal plant widely used in Asian countries, renowned for its effects of clearing heat, moistening the lungs, resolving phlegm, relieving cough, and alleviating asthma. In traditional medicinal practice, FUM is often used in combination with herbs such as Ephedra and Apricot Kernel. Among various preparation forms, decoction is the most common, and its traditional application methods hold significant reference value. Despite its extensive clinical application, the specific active components of FUM and its underlying mechanisms in treating asthma remain incompletely understood.

To evaluate the therapeutic effects of FUM extract, this study established an OVA-induced mouse asthma model in vivo. Additionally, an in vitro pyroptosis model was constructed by stimulating BEAS-2B cells with LPS and ATP.

This study established an in vivo mouse asthma model induced by OVA and constructed an in vitro cell pyroptosis model by stimulating BEAS-2B cells with LPS and ATP to evaluate the therapeutic effects of FUM extract. The prototype components of FUM absorbed into the bloodstream after administration were identified using liquid chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (LC/IM-QTOF-MS) combined with serum pharmacochemistry. Network pharmacology was employed to predict potential targets and pathways of the active components. The effects on relevant protein expression in both in vivo and in vitro models were validated through behavioral observation, pulmonary function testing, HE staining, immunohistochemistry, Western blotting, and immunofluorescence techniques. Molecular docking, molecular dynamics simulations, and cellular thermal shift assays were employed to predict or validate the interactions between key active components and core targets.

FUM extract significantly alleviated symptoms in OVA-induced asthmatic mice. LC/IM-QTOF-MS analysis identified nine prototype components of FUM absorbed into the bloodstream, mainly steroidal alkaloids. Through network pharmacology and experimental validation, FUM was found to inhibit the activation of the PI3K/AKT/NF-κB pathway in both mouse lung tissue and stimulated cells, while downregulating pyroptosis-related proteins (NLRP3, GSDMD, Caspase-1). Further molecular docking and validation experiments identified peiminine as the key active component, which demonstrated stable binding with the mTOR target.

This study investigated the active components and potential mechanisms of FUM in asthma using serum pharmacochemistry and systems pharmacology approaches. The novel findings reveal that FUM alleviates cellular pyroptosis and inflammatory responses by inhibiting the NLRP3 inflammasome, thereby exerting its therapeutic effects on asthma.