Zhang Peng, Li Zhipeng, Liang Minxue, Yuan Jinxian, Zhou Jinyu, Zhang Jie, Deng Jing, Chen Yalan, Tang Hao, Xu Chengjie, Shi Chenjun, Ma Limin, Liu Qiankun, Liu Yong, Chen Yangmei, Zhang Hui
Department of Neurology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China.
Department of Rehabilitation Sciences and Physiotherapy, University of Antwerp, Antwerp, 2610, Belgium.
Cell Commun Signal. 2025 Jul 8;23(1):327. doi: 10.1186/s12964-025-02239-3.
Despite continuous development of antiseizure medications (ASMs), global seizure control rates remain unsatisfactory, highlighting the urgent need for novel ASMs targeting distinct pathophysiological mechanisms. Inhibition of NLRP3 inflammasome activation represents an emerging strategy to simultaneously attenuate seizures and associated neuropsychiatric comorbidities. Therefore, this study investigates whether JC124, a novel NLRP3 inflammasome inhibitor, exerts neuroprotective effects in kainic acid (KA)-induced epileptic mice and in human induced pluripotent stem cell (hiPSC)-derived neurons stimulated by lipopolysaccharide (LPS) and adenosine triphosphate (ATP).
Summary-based Mendelian Randomization (SMR) was used to analyze the association between NLRP3 alleles and epilepsy susceptibility. NLRP3 knockout mice were generated, and then epileptic mice induced by intrahippocampal KA injection were administered JC124 (50 mg/kg, intraperitoneal) once daily for 28 days. The spontaneous recurrent seizures, hippocampal local field potential, depressive-like behavior, cognitive dysfunction, and locomotor ability of mice were evaluated. The brain tissues of the mice were collected for Western blotting, immunohistochemistry, immunofluorescence labeling, enzyme-linked immunosorbent assay, transmission electron microscopy, and morphological staining. The binding capacity of JC124 to the human NLRP3 protein was assessed using molecular docking and molecular dynamics simulations. hiPSC-derived neurons were used to explore the neuroprotective effects of JC124 against inflammatory injury in human neurons.
In this study, a positive correlation was identified between the expression of the NLRP3 gene and the susceptibility to epilepsy through SMR analysis. JC124 intervention markedly inhibited seizures and improved depressive-like behavior and cognitive dysfunction. It also reduced hippocampal neuronal loss, neuronal pyroptosis, microgliosis, and astrogliosis. Importantly, the neuroprotective effects of JC124 in KA-induced epileptic mice were mediated through the inhibition of the NLRP3 inflammasome. JC124 inhibited neuroinflammation and oxidative stress in KA-treated NLRP3 wild-type mice, but not in KA-treated NLRP3 knockout mice. Furthermore, JC124 bound directly to the human NLRP3 protein and alleviated neuroinflammation and oxidative stress in hiPSC-derived neurons stimulated by LPS and ATP.
These findings indicate that inhibiting the activation of the NLRP3 inflammasome with JC124 represents a potentially safe and innovative therapeutic strategy to mitigate neuroinflammation and oxidative stress in epilepsy and to alleviate seizures and associated neuropsychiatric comorbidities.
尽管抗癫痫药物（ASMs）不断发展，但全球癫痫控制率仍不尽人意，这凸显了迫切需要针对不同病理生理机制的新型抗癫痫药物。抑制NLRP3炎性小体激活是一种新兴策略，可同时减轻癫痫发作及相关神经精神共病。因此，本研究调查新型NLRP3炎性小体抑制剂JC124在海藻酸（KA）诱导的癫痫小鼠以及脂多糖（LPS）和三磷酸腺苷（ATP）刺激的人诱导多能干细胞（hiPSC）衍生神经元中是否发挥神经保护作用。
基于汇总的孟德尔随机化（SMR）用于分析NLRP3等位基因与癫痫易感性之间的关联。构建NLRP3基因敲除小鼠，然后对海马内注射KA诱导的癫痫小鼠每天腹腔注射一次JC124（50mg/kg），持续28天。评估小鼠的自发复发性癫痫发作、海马局部场电位、抑郁样行为、认知功能障碍和运动能力。收集小鼠脑组织进行蛋白质免疫印迹、免疫组织化学、免疫荧光标记、酶联免疫吸附测定、透射电子显微镜检查和形态学染色。使用分子对接和分子动力学模拟评估JC124与人NLRP3蛋白的结合能力。利用hiPSC衍生神经元探索JC124对人类神经元炎症损伤的神经保护作用。
在本研究中，通过SMR分析确定NLRP3基因表达与癫痫易感性之间存在正相关。JC124干预显著抑制癫痫发作，改善抑郁样行为和认知功能障碍。它还减少了海马神经元丢失、神经元焦亡、小胶质细胞增生和星形胶质细胞增生。重要的是，JC124在KA诱导的癫痫小鼠中的神经保护作用是通过抑制NLRP3炎性小体介导的。JC124抑制KA处理的NLRP3野生型小鼠中的神经炎症和氧化应激，但对KA处理的NLRP3基因敲除小鼠无效。此外，JC124直接与人NLRP3蛋白结合，并减轻LPS和ATP刺激的hiPSC衍生神经元中的神经炎症和氧化应激。
这些发现表明，用JC124抑制NLRP3炎性小体的激活是一种潜在安全且创新的治疗策略，可减轻癫痫中的神经炎症和氧化应激，并缓解癫痫发作及相关神经精神共病。
