Ferrostatin-1 inhibits ferroptosis and alleviates organophosphate nerve agent-induced cognitive deficits by regulating ACSL4/GPX4 and NCOA4/FTH1 pathways in the hippocampus of guinea pigs

Soman is a potent organophosphorus nerve agent (OPNA) that exerts its toxicity by irreversibly inhibiting acetylcholinesterase (AChE) via rapid aging. Nevertheless, the pathogenic mechanisms responsible for cognitive deficits following subacute soman exposure are poorly defined, representing a significant unmet medical need due to the absence of effective adjunctive treatments. Combined metabolomic and proteomic profiling reveals that ferroptosis-a regulated cell death process driven by lipid peroxidation-is implicated in cognitive decline following soman exposure. We developed a ferroptosis inhibitor Ferrostatin-1 (Fer-1, 2 mg/kg) treatment plan (2 mg/kg Fer-1 30 min before being subjected to soman challenge for 14 days) that effectively ameliorated soman-induced cognitive deficits and neuronal injury. Furthermore, our findings revealed that Fer-1 administration significantly suppressed AA mediated-neuroinflammatory and lipid peroxidation via ACSL4/GPX4 regulation, and restored redox imbalance by modulating the SLC7A11/GPX4 axis and glutamate-glutamine metabolism. Meanwhile, Fer-1 mitigated iron dyshomeostasis by attenuating NCOA4/FTH1-mediated ferritinophagy. Importantly, Fer-1 restored both mitochondrial respiration and dynamics. This was achieved by reversing soman-induced deficits in ATP synthesis and complexes I-III activity (linked to impaired Fe-S cluster biogenesis) and by regulating the expression of DRP1, OPA1, and MFN2 proteins. Finally, Pharmacodynamic evaluations demonstrated that both ACSL4 inhibitors and iron chelators effectively inhibited ferroptosis in the brain organoid model, suggesting their potential as neuroprotective agents. Collectively, our study demonstrated that subacute soman exposure-induced ferroptosis plays an important role in the development and progression of cognitive impairment, and inhibition of ferroptosis may be a potential therapeutic approach to prevent soman-induced cognitive impairment.