Verbascoside (VB) Inhibits Ischemic Stroke Progression via Suppression of Cell Damage and Ferroptosis By Targeting LCN2/ALOX5/SLC7A11/GPX4 Expression.

Ischemic stroke (IS), the most prevalent kind of stroke globally, poses a substantial threat to the health and economic well-being of the global population. Verbascoside (VB), a major bioactive constituent found in many medicinal plants, has demonstrated therapeutic potential in treating IS. Lipocalin 2 (LCN2) has been reported to accelerate the progression of IS. This study aimed to investigate the therapeutic effects and mechanism of VB in the context of IS. SK-N-SH cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to establish an in vitro cell model mimicking IS. Cellular behaviors were detected by CCK-8 assay, flow cytometry, and TUNEL. Inflammatory cytokines and markers associated with ferroptosis were measured using specific assay kits. Protein expression levels were examined by western blot. Additionally, molecular docking and cellular thermal shift assay (CETSA) were performed to analyze the binding between VB and LCN2. Besides, the BioGRID database, Co-immunoprecipitation (Co-IP), and immunofluorescence co-localization assays were employed to assess the interaction between LCN2 and arachidonate 5-lipoxygenase (ALOX5). Finally, the middle cerebral artery occlusion (MCAO) models were established for further analysis. OGD/R-induced SK-N-SH cell damage and ferroptosis were inhibited by VB treatment. Silencing LCN2 alleviated OGD/R-induced injury in SK-N-SH cells. VB modulated cell injury and ferroptosis by regulating LCN2 expression. An interaction was identified between LCN2 and ALOX5. Overexpression of arachidonate 5-lipoxygenase (ALOX5) reversed the protective effects of LCN2 silencing against OGD/R-induced cell damage. Additionally, VB was found to affect the expression of ALOX5 and the proteins in the solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) pathway. In vivo, VB suppressed the progression of IS. VB exerts its neuroprotective effects against stroke by inhibiting the expression of LCN2 and ALOX5, thereby activating the SLC7A11/GPX4 pathway and playing a crucial role in counteracting the pathological processes of stroke.