Neuronal Ferroptosis Attenuation by <em>RKIP</em> Inhibition Following Spontaneous Intracerebral Hemorrhage via <em>NRF2</em>/<em>HO-1</em> Pathway Activation

Raf kinase inhibitor protein (RKIP), a primary member of the phosphatidylethanolamine-binding protein family, is recognized for its involvement in various physiological processes, including cell differentiation, migration, cell cycle, and apoptosis. This study explored its function in neuronal ferroptosis following spontaneous intracerebral hemorrhage (ICH) and the associated mechanisms. A cellular model of spontaneous ICH was developed using cultured rat pheochromocytoma (PC12) cells stimulated with hemin. Cell viability, RNA expression, reactive oxygen species (ROS) levels, and lipid hydroperoxide (LPO) were assessed. Additionally, protein expression levels were measured. This study confirmed that RKIP inhibition is neuroprotective in vitro following ICH. This effect was associated with increased expression of glutathione peroxidase 4 (GPX4) and decreased expression of acyl-CoA synthetase long-chain family 4 (ACSL4), along with ROS and LPO levels in neurons. RKIP inhibition demonstrated a protective role against neuronal ferroptosis in vitro following ICH, potentially via activation of the nuclear factor E2-related factor 2/heme oxygenase-1 (NRF2/HO-1) pathway. This mechanism may offer insights into therapeutic strategies targeting neuronal ferroptosis in ICH.