Luteolin Attenuates Asthma Via Inhibiting ALOX15 Mediated Bronchial Epithelial Cell Ferroptosis

Ferroptosis, an iron-dependent form of cell death driven by lipid peroxidation, plays a critical role in the progression of asthma, yet its therapeutic modulation remains underexplored. Luteolin (Lut), a natural flavonoid known for its anti-inflammatory properties, presents a potential candidate for targeting ferroptosis in asthma. However, the precise mechanisms underlying its therapeutic effects are unclear. This study used a combination of bioinformatics, network pharmacology, molecular docking, and in vitro and in vivo experiments to investigate the role of luteolin in the treatment of ferroptosis in asthma. A collection of natural flavonoid compounds with high oral bioavailability and drug-like properties was assembled from the Chinese herbal medicine database. Through network pharmacology analysis, it was discovered that these flavonoids can effectively treat ferroptosis in asthma by regulating Arachidonic acid 15-lipoxygenase (ALOX15) and arachidonic acid metabolism. Further investigation through virtual docking revealed that luteolin is a key compound in the treatment of ferroptosis in asthma, targeting ALOX15. Molecular dynamics simulations demonstrated that the ALOX15-luteolin complex is stable and flexible. Cellular Thermal Shift Assay (CETSA) confirmed the thermal stability of luteolin and ALOX15, while Drug affinity response target stability (DARTS) assay showed that luteolin inhibits the proteolysis of ALOX15 by pronase. In in vitro experiments, it was observed that luteolin treatment reduced Fe²⁺ content and lipid ROS levels in a dose-dependent manner, while also downregulating ALOX15 and ACSL4 and upregulating SLC7A11 and GPX4, effectively alleviating ferroptosis induced by house dust mite (HDM) and lipopolysaccharide (LPS) in 16HBE cells. ALOX15 was specifically knocked down in 16HBE cells, and it was found that ALOX15 silencing and luteolin treatment could also inhibit ferroptosis in asthma. In vivo experiments and serum metabolomics analyses further confirmed that luteolin inhibits ferroptosis by suppressing ALOX15 expression and regulating arachidonic acid metabolism, ultimately alleviating asthma symptoms in mice. This study found that luteolin inhibited ALOX15-mediated ferroptosis in bronchial epithelial cells to alleviate asthma, highlighting the potential of luteolin as a promising therapeutic agent for asthma treatment.