Eupatilin mitigates ICH-induced brain injury via SOX2/SLC7A11 regulation of ferroptosis.

Intracerebral hemorrhage (ICH) is an acute and potentially fatal brain disease that causes severe brain damage, resulting in hemiplegia, speech disorders, and even coma or death. Recently, oxidative stress and ferroptosis are claimed involved in the pathogenesis of ICH brain injury. Eupatilin (EUP) is a flavonoid component with promising anti-inflammatory and anti-oxidative stress properties. The present study aimed to explore the pharmacological activity and potential mechanisms of EUP in treating ICH brain injury from the respect of ferroptosis. The ICH model was established in mouse, followed by treating with 3 and 10 mg/kg EUP. Elevated mNSS score, increased brain water content, reduced Nissl bodies, and enhanced EB concentrations were observed in ICH mice, which were sharply alleviated by EUP treatments. Furthermore, increased ROS, MDA, and iNOS levels; declined SOD activities; and enhanced proinflammatory factors like interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interleukin-1β (IL-1β) in ICH mice were remarkably reversed by EUP. In addition, increased iron⁺ cells and downregulated GPX4, FTH1, and SLC7A11, as well as reduced SOX2 levels, in ICH mice were rescued by EUP. SH-SY5Y cells were stimulated by hemin, followed by treating with EUP (25 and 50 μM). Enhanced ROS and MDA contents; decreased SOD activities; increased iron levels; and downregulated GPX4, FTH1, SLC7A11, and SOX2 were observed in hemin-stimulated SH-SY5Y cells, which were reversed by EUP. Moreover, silencing SOX2 abolished influences of EUP on oxidative stress and ferroptosis in hemin-stimulated SH-SY5Y cells. Collectively, EUP alleviated ICH-induced secondary brain injury by repressing SOX2/SLC7A11 axis-mediated ferroptosis, which provides potential treatment strategies for ICH in the clinic.