Giving old drugs new life: Disulfiram alleviates microglial pyroptosis and disulfidptosis-like cytoskeletal/mitochondrial alterations in cerebral ischemia-reperfusion injury

BACKGROUND: The cerebral ischemia-reperfusion injury (CIRI) induced by endovascular treatment represents a research challenge. Microglial pyroptosis plays a critical role in the neuroinflammatory damage associated with CIRI. Gasdermin D (GSDMD) is a key executor of pyroptosis, mediating plasma membrane rupture. Interestingly, research on pyroptosis has long focused on cell membrane rupture. We further hypothesize that cytoskeletal collapse during pyroptosis may facilitate cell rupture. This new perspective will contribute to a comprehensive understanding of the mechanisms of morphological changes during pyroptosis. Recent research has proposed disulfidptosis, an emerging form of cell death characterized by cytoskeletal collapse. Therefore, we speculated that disulfidptosis might occur concurrently with pyroptosis-associated cell rupture, and investigated whether disulfiram (DSL) could inhibit microglial pyroptosis and the associated disulfidptosis-related changes in a CIRI model. MATERIALS: Middle cerebral artery occlusion (MCAO) was established on rats, and the optimal dosage of DSL was determined. Neurological function, infarct volume, vascular diameter, cellular morphology, and key molecular mediators of microglial pyroptosis and disulfidptosis were tested. Furthermore, microglial mitochondrial fragmentation was investigated at the subcellular structural level. RESULTS: ) significantly improved neurological function in rats. Moreover, high-dose DSL reduced infarct size and attenuated cellular shrinkage in the ischemic penumbra. GSDMD and SLC7A11 were co-localized in microglia. DSL suppressed the expression of pyroptosis-related proteins, including GSDMD-N-terminal, caspase-1 p10, and cleaved caspase-11. In addition, DSL mitigated SLC7A11 loss and inhibited the expression of NOX4 and ARP2. At the mitochondrial level, DSL alleviated microglial mitochondrial fragmentation and downregulated the expression of DRP1 and VDAC1. CONCLUSION: Following CIRI, disulfiram suppressed pyroptosis by inhibiting the caspase-1/11-GSDMD pathway and attenuated disulfidptosis-like changes via modulation of the SLC7A11-NOX4-ARP2 axis, suggesting its potential neuroprotective effect.