Icariside II preconditioning evokes robust neuroprotection against ischaemic stroke, by targeting Nrf2 and the OXPHOS/NF‐κB/ferroptosis pathway

Background and Purpose Astrocytic nuclear factor erythroid‐derived 2‐related factor 2 (Nrf2) is a potential therapeutic target of ischaemic preconditioning (IPC). Icariside II (ICS II) is a naturally occurring flavonoid derived from Herba Epimedii with Nrf2 induction potency. This study was designed to clarify if exposure to ICS II mimicks IPC neuroprotection and if Nrf2 from astrocytes contributes to ICS II preconditioning against ischaemic stroke. Experimental Approach Mice with transient middle cerebral artery occlusion (MCAO)‐induced focal cerebral ischaemia and primary astrocytes challenged with oxygen–glucose deprivation (OGD) were used to explore the neuroprotective effect of ICS II preconditioning. Additionally, Nrf2‐deficient mice were pretreated with ICS II to determine whether ICS II exerts its neuroprotection by activating Nrf2. Key Results ICS II pretreatment mitigated cerebral injury in the mouse model of ischaemic stroke along with improving long‐term recovery. Furthermore, proteomics screening identified Nrf2 as a crucial gene evoked by ICS II treatment and required for the anti‐oxidative effect and anti‐inflammatory effect of ICS II. Also, ICS II directly bound to Nrf2 and reinforced the transcriptional activity of Nrf2 after MCAO. Moreover, ICS II pretreatment exerted cytoprotective effects on astrocyte cultures following lethal OGD exposure, by promoting Nrf2 nuclear translocation and activating the OXPHOS/NF‐κB/ferroptosis axis, while neuroprotection was decreased in Nrf2‐deficient mice and Nrf2 siRNA blocked effects of ICS II. Conclusion and Implications ICS II preconditioning provides robust neuroprotection against ischaemic stroke via the astrocytic Nrf2‐mediated OXPHOS/NF‐κB/ferroptosis axis. Thus, ICS II could be a promising Nrf2 activator to treat ischaemic stroke.