Integrated network pharmacology, molecular docking, and experimental validation elucidate the anti-inflammatory and antioxidant mechanisms of apigenin in LPS-induced acute lung injury.

Abstract

Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), are associated with high clinical mortality and lack effective therapeutic agents. The natural flavonoid apigenin possesses well-defined anti-inflammatory and antioxidant activities; however, its protective mechanism in ALI remains to be systematically elucidated. In this study, we established LPS-induced mouse models of ALI and BEAS-2B human bronchial epithelial cell injury models, combined with network pharmacology, molecular docking, and 100 ns molecular dynamics simulations, and employed the ferroptosis inhibitor Fer-1 and inducer Erastin for mechanistic validation, to comprehensively evaluate the protective effects of apigenin. Our results demonstrated that apigenin dose-dependently alleviated pulmonary histopathological damage, reduced inflammatory cell infiltration, myeloperoxidase activity, and the levels of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α. Concurrently, apigenin inhibited the phosphorylation of NF-κB and JAK2-STAT3 pathways, upregulated the expression of GPX4 and SLC7A11, decreased Fe²⁺ and malondialdehyde levels, and attenuated lipid peroxidation. These effects were similar to those of Fer-1 and were partially reversed by Erastin. Network pharmacology and molecular simulations revealed that apigenin stably binds to core targets including MMP9, EGFR, and ESR1, and KEGG enrichment analysis significantly pointed to the NF-κB and JAK-STAT pathways. Collectively, apigenin effectively alleviates LPS-induced ALI through coordinated regulation of the NF-κB/JAK2-STAT3 pathway and inhibition of inflammatory responses, ferroptosis, and oxidative stress, thus providing a novel theoretical basis and a candidate therapeutic strategy for the treatment of ALI with this flavonoid.