Abstract: Epilepsy is a prevalent neurological disorder in which hippocampal neuronal damage, particularly ferroptosis, plays a critical role. Previous studies have shown that hypoxia-inducible factor 1α is considered an important regulator of cellular stress responses and has been confirmed to play a critical role in the occurrence of various diseases. However, the mechanisms by which hypoxia-inducible factor 1α is related to epilepsy and neuronal ferroptosis remain unclear. In this study, we used a pentylentetrazole-induced chronic epilepsy mouse model and treated the mice with intraperitoneal administration of PX-478, a hypoxia-inducible factor-1α inhibitor. Our results showed that PX-478 significantly prolonged the latency of epilepsy, reduced seizure severity, and shortened seizure duration. PX-478 also alleviated neuronal damage in the hippocampal CA1 and CA2 regions, reduced levels of reactive oxygen species and malondialdehyde, and increased levels of superoxide dismutase, catalase, and glutathione peroxidase. Transmission electron microscopy showed that PX-478 treatment reduced mitochondrial damage in the hippocampal neurons of epileptic mice, and significantly improved mitochondrial length and area. Additionally, PX-478 preferentially reduced Fe2+ levels and the expression of cyclooxygenase-2, ferritin heavy chain 1 and transferrin in the hippocampus of epileptic mice. It also inhibited the activity of the hypoxia-inducible factor 1α/heme oxygenase-1 pathway. In summary, these findings suggest that PX-478 has the potential to treat epilepsy by inhibiting the hypoxia-inducible factor 1α/heme oxygenase-1 pathway, alleviating oxidative stress, and reducing ferroptosis in hippocampal neurons. 

Key words: epilepsy, ferroptosis, heme oxygenase-1, hippocampus, hypoxia-inducible factor 1α, mitochondrial ultrastructure, oxidative stress, PX-478, reactive oxygen species, seizure behavior