A ferulic acid derivative FAD012 protects brain microvascular endothelial cells from H₂O₂-induced ferroptosis via...

Ferroptosis, a regulated form of necrotic cell death characterized by iron-dependent lipid peroxidation, has been implicated in blood-brain barrier (BBB) disruption during ischemia-reperfusion injury, particularly in brain microvascular endothelial cells. We previously developed a novel derivative of ferulic acid (FA), FAD012, and demonstrated its neurovascular protective effects in multiple rat models of cerebral ischemia. In this study, we investigated whether FAD012 protects rat brain microvascular endothelial cells (RBMVECs) from hydrogen peroxide (H₂O₂)-induced ferroptosis and further elucidated its underlying mechanisms. H₂O₂-induced cell death was attenuated by ferroptosis inhibitors (ferrostatin-1 and deferoxamine) and was accompanied by downregulation of glutathione peroxidase 4 and 4-hydroxynonenal accumulation, collectively indicating the induction of ferroptosis. Pretreatment with FAD012 restored cell viability, mitigated lipid peroxidation, and prevented ferroptosis more effectively than its parent compound, FA. Mechanistically, FAD012 scavenged reactive oxygen species and promoted nuclear factor erythroid 2-related factor 2 (NRF2) nuclear translocation and downstream antioxidant signaling. Inhibition of NRF2 by ML385 abolished the cytoprotective effects of FAD012, confirming the critical role of NRF2 activation. These findings suggest that FAD012 suppresses H₂O₂-induced ferroptosis in RBMVECs through both direct antioxidant activity and NRF2 activation, providing a mechanistic basis for its potential to preserve BBB integrity under oxidative stress in vivo.