Traditional Chinese Herbal Formula XuMingZhuSan Alleviates Neuronal Ferroptosis in Rats After Stroke by Activating the NRF2/GPX4/SLC7A11 Pathway.

This paper reported the efficacy of Traditional Chinese herbal formula XuMingZhuSan (XMZS) in stroke. Middle cerebral artery occlusion (MCAO) rat model was constructed and treated by XMZS and ML385. Brain sections of rats underwent histologic staining methods to assess brain infarction, neuronal and mitochondrial damage, and reactive oxygen species (ROS) production. Oxygen-glucose deprivation/reperfusion (OGD/R)-induced hippocampal neurons were treated with XMZS and ML385. Apoptosis, ROS, mitochondrial membrane potential, and lipid peroxidation in hippocampal neurons were monitored by TUNEL, DCFH-DA, JC-1, and C11-Bodipy staining. Ferroptosis-related indicators in brains and hippocampal neurons were assayed using commercial kits. NRF2/GPX4/SLC7A11 pathway activity within brains and hippocampal neurons was assessed by Western blotting. In MCAO rats, XMZS improved neurological function; reduced cerebral infarction volume; relieved neuronal damage in the cortex and hippocampal CA1 and CA3 regions, and mitochondrial damage within the brain; suppressed ROS in the cortex and hippocampal CA1 and CA3 regions; decreased Fe and MDA contents within the brain; increased SOD and GSH within the brain; and activated NRF2/GPX4/SLC7A11 pathway within the brain. These influences induced by XMZS on MCAO rats were eliminated by ML385. XMZS mitigated apoptosis of OGD/R-induced hippocampal neurons; reduced LDH activity and ROS; declined Fe and MDA contents; raised SOD and GSH levels; elevated mitochondrial membrane potential; suppressed lipid peroxidation; and activated NRF2/GPX4/SLC7A11 pathway. ML385 reversed these effects of XMZS on the OGD/R-induced hippocampal neurons. XMZS may activate NRF2/GPX4/SLC7A11 pathway to treat stroke by attenuating neuronal ferroptosis and is applicable in clinical management of stroke.