Puerarin Alleviates Cerebral Ischemia-Reperfusion Injury by Inhibiting Ferroptosis Through SLC7A11/GPX4/ACSL4 Axis and Alleviate Pyroptosis Through Caspase-1/GSDMD Axis.

Cerebral ischemia-reperfusion (CIRI) represents a complex disease entity that encompasses multiple pathways. The occurrence of CIRI induces cerebral infarction, accompanied by brain tissue necrosis and focal neuronal impairment. Previous studies have demonstrated that ferroptosis, a specific cell death pathway implicated in CIRI, plays a crucial role in mediating the pathophysiological process of this condition. Puerarin, is known to possess vasodilatory, antioxidant, and neuroprotective properties. However, its precise role in ferroptosis as well as the underlying mechanisms remains elusive. In this study, we delved into the neuroprotective mechanisms of puerarin using both the rat middle cerebral artery occlusion (MCAO) model and the HT22 cell model of oxygen-glucose deprivation/reperfusion (OGD/R). In the MCAO model, puerarin was found to exhibit an inhibitory effect on ACSL4, which was consistent with that of rosiglitazone. Simultaneously, it was capable of counteracting the inhibition of GPX4 by RSL3. These findings suggest that puerarin modulates GPX4 and ACSL4, thereby exerting an inhibitory effect on ferroptosis. The ferroptosis-protective effect of puerarin was further corroborated in the OGD/R through a positive control experiment with ferrostatin-1, a lipid peroxidation inhibitor. Furthermore, we also recognized the importance of other cell death modalities, such as pyroptosis. Consequently, we verified the neuroprotective effect of puerarin by examining the influence of caspase-1 and GSDMD in HT22. Mechanistically, puerarin alleviates CIRI by respectively inhibiting ferroptosis through the SLC7A11/GPX4/ACSL4 axis and pyroptosis through the caspase-1/GSDMD axis. This research provides novel insights into the targeting and therapeutic potential of puerarin for the treatment of CIRI.