Fujian Key Laboratory of Translational Research in Cancer and Neurodegenerative Diseases, Institute for Translational Medicine, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, China.
Fujian Provincial Key Laboratory of Natural Medicine Pharmacology, Institute of Materia Medica, School of Pharmacy, Fujian Medical University, Fuzhou 350122, China.
Int J Mol Sci. 2022 Jun 7;23(12):6370. doi: 10.3390/ijms23126370.
Glutamate excitotoxicity induces neuronal cell death during epileptic seizures. Death-associated protein kinase 1 (DAPK1) expression is highly increased in the brains of epilepsy patients; however, the underlying mechanisms by which DAPK1 influences neuronal injury and its therapeutic effect on glutamate excitotoxicity have not been determined. We assessed multiple electroencephalograms and seizure grades and performed biochemical and cell death analyses with cellular and animal models. We applied small molecules and peptides and knocked out and mutated genes to evaluate the therapeutic efficacy of kainic acid (KA), an analog of glutamate-induced neuronal damage. KA administration increased DAPK1 activity by promoting its phosphorylation by activated extracellular signal-regulated kinase (ERK). DAPK1 activation increased seizure severity and neuronal cell death in mice. Selective ERK antagonist treatment, DAPK1 gene ablation, and uncoupling of DAPK1 and ERK peptides led to potent anti-seizure and anti-apoptotic effects in vitro and in vivo. Moreover, a DAPK1 phosphorylation-deficient mutant alleviated glutamate-induced neuronal apoptosis. These results provide novel insight into the pathogenesis of epilepsy and indicate that targeting DAPK1 may be a potential therapeutic strategy for treating epilepsy.
谷氨酸兴奋性毒性在癫痫发作期间诱导神经元细胞死亡。在癫痫患者的大脑中，死亡相关蛋白激酶 1（DAPK1）的表达高度增加；然而，DAPK1 影响神经元损伤的潜在机制及其对谷氨酸兴奋性毒性的治疗效果尚未确定。我们评估了多种脑电图和癫痫发作等级，并使用细胞和动物模型进行了生化和细胞死亡分析。我们应用小分子和肽，并敲除和突变基因来评估作为谷氨酸诱导的神经元损伤类似物的海人酸（KA）的治疗功效。KA 给药通过促进其被激活的细胞外信号调节激酶（ERK）磷酸化来增加 DAPK1 活性。DAPK1 的激活增加了小鼠的癫痫发作严重程度和神经元细胞死亡。选择性 ERK 拮抗剂处理、DAPK1 基因缺失以及 DAPK1 和 ERK 肽的解偶联导致体外和体内具有强大的抗癫痫和抗细胞凋亡作用。此外，DAPK1 磷酸化缺陷型突变减轻了谷氨酸诱导的神经元凋亡。这些结果为癫痫的发病机制提供了新的见解，并表明靶向 DAPK1 可能是治疗癫痫的一种潜在治疗策略。
