(S)-SA, a bioactive flavonoid from Dalbergiae Odoriferae Lignum, alleviates cerebral ischemia-reperfusion injury via SLC11A2-mediated ferroptosis inhibition.

BACKGROUND: Cerebral ischemia-reperfusion injury (CIRI) remains a major barrier to neurological recovery after ischemic stroke, and effective pharmacological interventions targeting upstream neurovascular unit mechanisms related to ferroptosis and blood-brain barrier (BBB) injury are limited. PURPOSE: This study aimed to clarify the protective effects of Dalbergiae Odoriferae Lignum (DOL) and its active constituent sativanone (SA), particularly (S)-SA, against CIRI and to define the direct target and mechanism underlying its activity. STUDY DESIGN AND METHODS: The protective effects of DOL, SA, and (S)-SA were evaluated in a middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model and an oxygen-glucose deprivation/reoxygenation (OGD/R) hcMEC/D3 cell model. Brain-distributed constituents were analyzed by DESI-MS and activity screening. Direct target identification and validation were performed using activity-based protein profiling (ABPP), a proteolysis-targeting chimera (PROTAC)-based targeted degradation strategy, cellular thermal shift assay (CETSA), drug affinity responsive target stability (DARTS), surface plasmon resonance (SPR), molecular docking and molecular dynamics simulations, and target-dependency studies. RESULTS: DOL protected against MCAO/R-induced CIRI, and SA was detected in brain tissue and showed protective activity in vitro and in vivo. Among the SA enantiomers, (S)-SA exerted stronger protective effects than (R)-SA. Orthogonal chemical biology and target validation identified SLC11A2 as a direct functional target of (S)-SA. Mechanistically, (S)-SA inhibited SLC11A2-mediated Fe²⁺ uptake, reduced ferroptosis-related lipid peroxidation and oxidative injury, attenuated inflammatory signaling, and preserved BBB integrity. CONCLUSION: These findings indicate that (S)-SA alleviates CIRI by targeting SLC11A2-mediated Fe²⁺ uptake and ferroptosis, supporting (S)-SA as a potential lead compound for CIRI therapy.