Loki Zupa alleviates type 2-high acute asthma airway inflammation by regulating the glutathione metabolism-ferroptosis axis.

BACKGROUND: Oxidative stress-induced disruption of glutathione metabolism triggers ferroptosis in airway epithelial cells, thereby exacerbating type 2-high acute asthma. Loki Zupa (LKZP) is a classic Uyghur formula used for asthma with confirmed anti-inflammatory and antioxidant effects, but its mechanism via the glutathione-ferroptosis axis remains unclear. PURPOSE: This study aimed to characterize LKZP's efficacy against type 2-high acute asthma and to elucidate its mechanism, focusing on the glutathione metabolism-ferroptosis axis. METHODS: An OVA-induced type 2-high acute asthma mouse model was established to evaluate the anti-inflammatory and antioxidant effects of LKZP. UHPLCHRMS was used to identify its active components, and network pharmacology combined with proteomics and metabolomics was applied to predict the underlying mechanisms. An RSL3-induced ferroptosis cell model was employed to verify LKZP's protective effects on cell viability, glutathione (GSH) and malondialdehyde (MDA) levels, as well as the expression of ferroptosis-related proteins. Molecular docking and surface plasmon resonance (SPR) assay were performed to collectively evaluated the binding affinity between Linarin (an active component of LKZP) and GSTM1/GSTM2. RESULTS: LKZP significantly alleviated airway inflammation and oxidative stress injury in asthmatic mice. Its anti-asthmatic effect was linked to the glutathione metabolism-ferroptosis axis. In vivo and in vitro experiments confirmed that LKZP restored glutathione metabolic homeostasis, scavenged excessive reactive oxygen species (ROS), reduced iron overload and lipid peroxidation, and inhibited ferroptosis in lung tissue and airway epithelial cells. Molecular docking and molecular dynamics simulation predicted stable binding of Linarin to GSTM1 and GSTM2. SPR analysis further confirmed these interactions and demonstrated a higher binding affinity of Linarin toward GSTM1 than GSTM2. CONCLUSION: LKZP alleviated airway inflammation and oxidative stress in type 2-high acute asthma by regulating the glutathione metabolism-ferroptosis axis, providing novel mechanistic insights for its asthma therapy.