Department of Neurology, The Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen, 518033, People's Republic of China.
Department of Neurology, School of Medicine, Guangzhou First People's Hospital, South China University of Technology, Guangzhou, 510180, Guangdong Province, People's Republic of China.
Neurochem Res. 2023 Nov;48(11):3349-3362. doi: 10.1007/s11064-023-03950-8. Epub 2023 Jun 2.
Uncovering mechanisms underlying epileptogenesis aids in preventing further epilepsy progression and to lessen seizure severity and frequency. The purpose of this study is to explore the antiepileptogenic and neuroprotective mechanisms of EGR1 in neuron injuries encountered in epilepsy. Bioinformatics analysis was conducted to identify the key genes related to epilepsy. The mice were rendered epileptic using the kainic acid protocol, followed by measurement of seizure severity, high amplitude and frequency, pathological changes of hippocampal tissues and neuron apoptosis. Furthermore, an in vitro epilepsy model was constructed in the neurons isolated from newborn mice, which was then subjected to loss- and gain-of-function investigations, followed by neuron injury and apoptosis assessment. Interactions among EGR1, METTL3, and VIM were analyzed by a series of mechanistic experiments. In the mouse and cell models of epilepsy, VIM was robustly induced. However, its knockdown reduced hippocampal neuron injury and apoptosis. Meanwhile, VIM knockdown decreased inflammatory response and neuron apoptosis in vivo. Mechanistic investigations indicated that EGR1 transcriptionally activated METTL3, which in turn downregulated VIM expression through m6A modification. EGR1 activated METTL3 and reduced VIM expression, thereby impairing hippocampal neuron injury and apoptosis, preventing epilepsy progression. Taken together, this study demonstrates that EGR1 alleviates neuron injuries in epilepsy by inducing METTL3-mediated inhibition of VIM, which provides clues for the development of novel antiepileptic treatments.
揭示癫痫发生的机制有助于防止癫痫进一步进展，减轻发作的严重程度和频率。本研究旨在探讨 EGR1 在癫痫中神经元损伤中的抗癫痫发生和神经保护机制。通过生物信息学分析鉴定与癫痫相关的关键基因。使用海人酸方案使小鼠癫痫发作，然后测量癫痫发作的严重程度、高振幅和高频率、海马组织的病理变化和神经元凋亡。此外，在从新生小鼠中分离的神经元中构建体外癫痫模型，然后进行缺失和功能获得研究，随后评估神经元损伤和凋亡。通过一系列机制实验分析 EGR1、METTL3 和 VIM 之间的相互作用。在癫痫的小鼠和细胞模型中，VIM 被强烈诱导。然而，其敲低减少了海马神经元损伤和凋亡。同时，VIM 敲低减少了体内的炎症反应和神经元凋亡。机制研究表明，EGR1 转录激活 METTL3，METTL3 通过 m6A 修饰反过来下调 VIM 的表达。EGR1 激活 METTL3 并降低 VIM 表达，从而损害海马神经元损伤和凋亡，防止癫痫进展。总之，本研究表明，EGR1 通过诱导 METTL3 介导的 VIM 抑制来减轻癫痫中的神经元损伤，为开发新的抗癫痫治疗方法提供了线索。
