Yang Pu, Yu Qian
Department of Neurology, The Third Xiangya Hospital of Central South University, Changsha, Hunan, People's Republic of China.
Department of Hematology, The Second Xiangya Hospital of Central South University, Changsha, Hunan, People's Republic of China.
FASEB J. 2025 Sep 15;39(17):e70987. doi: 10.1096/fj.202500188R.
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 Mg-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 Mg-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 Mg-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.
癫痫是一种常见的威胁人类健康的慢性神经系统疾病。然而，FOXC1在癫痫中的作用及其与细胞焦亡的关系尚未得到充分研究。获取Sprague-Dawley大鼠以构建颞叶癫痫（TLE）模型。用无镁培养基处理SH-SY5Y细胞以诱导体外细胞模型。采用苏木精-伊红染色、金属硫蛋白染色和尼氏染色进行组织学评估。用流式细胞术检测细胞焦亡。采用酶联免疫吸附测定法检测炎性细胞因子水平。进行免疫组织化学和免疫荧光以确定基因的表达和分布。进行染色质免疫沉淀、RNA免疫沉淀、甲基化RNA免疫沉淀和双荧光素酶报告基因分析以研究分子间相互作用。使用逆转录定量聚合酶链反应和蛋白质免疫印迹法检测基因的mRNA和蛋白质水平表达。FOXC1在TLE大鼠海马组织中低表达，FOXC1的过表达减轻了神经元损伤。FOXC1的过表达通过抑制无镁刺激的SH-SY5Y细胞中NLRP3介导的细胞焦亡，下调了NLRP3、半胱天冬酶1以及白细胞介素-1β和白细胞介素-18的水平。此外，FOXC1与NLRP3启动子结合并抑制NLRP3转录。发现FOXC1受METTL3/YTHDF1轴介导的m6A甲基化调控。METTL3的过表达通过YTHDF1依赖性调控FOXC1减轻了无镁诱导的神经元损伤和细胞焦亡。总的来说，这些数据证实FOXC1通过转录抑制NLRP3抑制癫痫中的神经元损伤和细胞焦亡，这可能与METTL3/YTHDF1轴介导的m6A甲基化有关。
