Combined network pharmacology and experimental validation reveals the mechanism of Gentiana veitchiorum Hemsl. and its active ingredient in the treatment of acute lung injury.

Gentiana veitchiorum Hemsl. is the principal species included in "Bang Jian", a Tibetan term referring to several Gentiana plants. It has been widely used in clinical practice for the treatment of respiratory diseases such as cough, acute and chronic bronchitis, and pneumonia. Severe bronchitis and pneumonia can precipitate acute lung injury (ALI) through the propagation of inflammation or worsening infection. As a common and severe critical condition, ALI is characterized by high morbidity and mortality. However, the active constituents and underlying mechanism of G. veitchiorum in the treatment of ALI remain unclear. In this study, a lipopolysaccharide (LPS)-induced ALI mouse model was established to evaluate the therapeutic effects of G. veitchiorum extract on ALI. Network pharmacology and molecular docking were employed to identify potential active compounds and elucidate the molecular mechanisms involved. Both in vivo and in vitro experiments were then performed to validate the pharmacological efficacy and mechanisms of the extracts and principal active compounds. Preliminary pharmacodynamic experiments showed that G. veitchiorum effectively ameliorated LPS-induced ALI in mice, likely by alleviating pulmonary edema, mitigating inflammatory infiltration and diffuse hemorrhage in the lung tissue, and regulating oxidative stress. Network pharmacology and molecular docking further identified swertiamarin (SWE) as the key active compound responsible for the therapeutic effects against ALI. In vivo experiments showed that SWE significantly reduced lung edema, suppressed the expression of inflammatory factors, restored oxidative stress-related enzyme activity, regulated macrophage polarization, and downregulated both the expression and phosphorylation of key proteins, thereby inhibiting activation of the TLR4/NF-κB/PI3K signaling pathway. In vitro experiments further confirmed that SWE significantly inhibited reactive oxygen species (ROS) generation as well as the secretion and mRNA expression of inflammatory cytokines. Inhibition of TLR4 and MyD88 protein expression, along with phosphorylation of p65, AKT and PI3K, as well as suppression of the nuclear translocation of p65 and p-AKT, collectively contributed to the blockade of NF-κB activation. These results suggest that G. veitchiorum and its active ingredients exert anti-ALI effects primarily through the inhibition of inflammatory responses, attenuation of oxidative stress, and modulation of the TLR4/NF-κB/PI3K signaling pathway.