Botulinum Toxin Type A as a Therapeutic Agent in Epilepsy: Attenuation of Neuronal Ferroptosis and Cognitive Dysfunction.

[PURPOSE] Epilepsy is one of the most common neurological disorders with no effective drugs to prevent seizures or their progression. Iron modulation is a potential advanced treatment for seizures. We aim to investigate whether botulinum toxin type A (BoNT/A) can attenuate epilepsy-induced neuronal death and maintain cognitive function by inhibiting ferroptosis.

[METHOD] We established an epileptic rat model and intervened with BoNT/A to assess its influence on cognitive functions and the pathological damage of hippocampal tissues. Rat hippocampal neuronal cells were treated with magnesium-free induction solution to establish an epileptic cell model and intervened using BoNT/A. Changes in ferrous ions (Fe), malondialdehyde (MDA), and glutathione (GSH) were detected in hippocampal tissues and cells. Western blot (WB) and RT-qPCR were used to detect the protein expression of the iron death markers, including GPX4, ACSL4, and SLC7A11.

[FINDING] We found that BoNT/A attenuated epileptiform behavior and cognitive deficits and ameliorated hippocampal tissue damage in rats under lithium chloride-pilocarpine-induced epilepsy. In vitro BoNT/A treatment exerted potent neuroprotective effects on hippocampal neuronal cells treated by magnesium-free induction solution. These protective effects were related to the regulation of ferroptosis mediated by the GPX4/ACSL4/SLC7A11 proteins.

[CONCLUSION] These results suggest that BoNT/A is effective in preventing epileptic neuronal iron death and attenuates cognitive dysfunction through the ferroptosis pathway.