Icaritin alleviates cerebral ischemia-reperfusion injury by reducing ferroptosis via inhibiting excessive mPTP opening

The narrow therapeutic window of reperfusion therapy highlight the urgent need for novel neuroprotective strategies against ischemic stroke, wherein ferroptosis is a key contributor to neuronal death in the ischemic penumbra. This study demonstrates that icaritin (ICT) treatment significantly improved functional outcomes, reduced infarct size,and enhanced neuronal viability in middle cerebral artery occlusion (MCAO) rats and oxygen-glucose deprivation and reoxygenation (OGD/R) PC-12 cells. Transcriptomic and biochemical analyses confirmed that ICT’s neuroprotection was associated with the suppression of ferroptosis. Mechanistically, ICT inhibited pathological mitochondrial permeability transition pore (mPTP) opening, thereby preserving mitochondrial membrane potential and attenuating reactive oxygen species (ROS) production. This action was essential, as the protective effect was mimicked by cyclosporin A (CsA) and abolished by lonidamine (LND). Molecular docking revealed stable binding of ICT to voltage-dependent anion channel (VDAC) and cyclophilin D (CypD.) In conclusion, ICT confers neuroprotection by inhibiting mPTP excessive opening, which in turn blocks the downstream ferroptosis cascade. This work elucidates a detailed mechanism for ICT and validates the “mPTP-ferroptosis” axis as a promising therapeutic target for ischemic stroke.