N6022 Alleviates Cerebral Ischemia/Reperfusion Injury by Attenuating Ferroptosis: A Potential Therapeutic Target for Ischemic Stroke

However, the protective role of N6022 in ischemic stroke and its specific mechanism of action remain unclear. This study aimed to investigate whether N6022 could alleviate cerebral ischemia/reperfusion (I/R) injury by attenuating ferroptosis and explore the underlying mechanisms contributing to its positive effects. The goal was to uncover potential molecular medicine for the clinical treatment of ischemic stroke. To simulate human cerebral I/R processes, we established an MCAO/R mouse model and an OGD/R cell model.

Both in vitro and in vivo experiments were conducted to observe the effects of N6022 on MCAO/R-induced brain injury, neurological deficits, apoptosis, and ferroptosis in mice, as well as OGD/R-induced BV2 cell injury, apoptosis, and ferroptosis. The results demonstrated that N6022 effectively protected against MCAO/R-induced brain damage and neurological deficits in mice, as well as OGD/R-induced BV2 cell damage.

Mechanistically, N6022 promoted the nuclear translocation of Nrf2, leading to increased expression of GPX4 and SLC7A11. This, in turn, enhanced the intracellular antioxidant capacity and attenuated ferroptosis. The specific Nrf2 inhibitor ML385 partially reversed the protective effects of N6022.

Furthermore, N6022 inhibited GSNOR, which disrupted its interaction with GSTP1. As a result, the antioxidant capacity of GSTP1 was increased, leading to the attenuation of ferroptosis. This effect was blocked by the specific knockdown of GSTP1.

Overall, these findings suggest that N6022 exerts protective effects in ischemic stroke by attenuating ferroptosis. The mechanisms involve the promotion of Nrf2 nuclear translocation, upregulation of GPX4 and SLC7A11, and interference with the GSNOR-GSTP1 interaction. These findings provide valuable insights into the potential molecular targets for the clinical treatment of ischemic stroke.