Hspb1 inhibits microglial ferroptosis and pro-inflammatory activation to alleviate cerebral ischemia/reperfusion injury in mice.

Heat shock protein beta-1 may be involved in regulating ferroptosis in cells. The expression of heat shock protein beta-1 is upregulated after stroke; however, the underlying mechanism of action of heat shock protein beta-1 in cerebral ischemia/reperfusion injury remains unclear. Here, using both in vivo and in vitro models of ischemic injury—middle cerebral artery occlusion/reperfusion in C57BL/6J mice and oxygen-glucose deprivation/reoxygenation in BV-2 microglial cells—we observed that heat shock protein beta-1 overexpression significantly reduced infarct volume, mitigated neuronal loss, and improved neurological outcomes. Mechanistically, heat shock protein beta-1 attenuated lipid peroxidation, intracellular iron accumulation, and reactive oxygen species generation in microglia; this was accompanied by enhanced glutathione peroxidase 4 expression and suppressed nuclear factor-κB pathway activation. Notably, the pharmacological activation of nuclear factor-κB with phorbol 12-myristate 13-acetate reversed the protective effects of heat shock protein beta-1, confirming the functional relevance of this pathway. Together, our findings indicate that heat shock protein beta-1 exerts neuroprotective effects against cerebral ischemia/reperfusion injury by suppressing microglial ferroptosis and pro-inflammatory activation via modulation of the nuclear factor-κB/glutathione peroxidase 4 signaling axis. These findings establish heat shock protein beta-1 as a critical regulator of the nuclear factor-κB/glutathione peroxidase 4 axis in microglia, thereby offering a dual-targeted strategy to inhibit ferroptosis and inflammation in ischemic stroke. Importantly, our study highlights heat shock protein beta-1 as a promising therapeutic candidate for preserving neurological function following cerebral ischemic injury.