Downregulation of FOXC1 Modified by METTL3/YTHDF1 Axis Aggravates NLRP3-Mediated Inflammasomes Formation and Cell Pyroptosis in Epilepsy

Abstract

Epilepsy is a common chronic nervous system disease that threatens human health. However, the role of FOXC1 and its relations with pyroptosis have not been fully studied in epilepsy. Sprague–Dawley rats were obtained for constructing temporal lobe epilepsy (TLE) models. SH-SY5Y cells were treated with Mg²⁺-free medium for inducing in vitro cell models. H&E, TIMM, and Nissl staining were employed for histological evaluation. Flow cytometry was used for measuring cell pyroptosis. ELISA was implemented for detecting the levels of inflammatory cytokines. Immunohistochemistry and immunofluorescence were performed to determine the expression and distribution of genes. ChIP, RIP, MeRIP, and dual-luciferase reporter analysis were performed to investigate the intermolecular interactions. The mRNA and protein level expression of genes were examined using RT-qPCR and western blotting. FOXC1 was lowly expressed in hippocampus tissue of TLE rats, and overexpression of FOXC1 alleviated neuronal damage. Overexpression of FOXC1 downregulated the levels of NLRP3, caspase 1, as well as IL-1β and IL-18 by inhibiting NLRP3-mediated pyroptosis in Mg²⁺-free–stimulated SH-SY5Y cells. Furthermore, FOXC1 bound to the promoter of NLRP3 and inhibited NLRP3 transcription. FOXC1 was found to be regulated by the METTL3/YTHDF1 axis-mediated m6A methylation. Overexpression of METTL3 attenuated Mg²⁺-free-induced neuronal injury and pyroptosis through YTHDF1-dependent regulation of FOXC1. Collectively, these data confirmed that FOXC1 suppressed neuronal injury and pyroptosis in epilepsy by transcriptionally inhibiting NLRP3, which might be correlated to METTL3/YTHDF1 axis-mediated m6A methylation.