Song Honghong, Li Yi, Liu Jiayu, Bai Xianshu, Jiang Li, Li Tingsong
Department of Neurology Children's Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders Ministry of Education Key Laboratory of Child Development and Disorders Chongqing Key Laboratory of Child Neurodevelopment and Cognitive Disorders Chongqing China.
Department of Rehabilitation Children's Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders Ministry of Education Key Laboratory of Child Development and Disorders Chongqing Key Laboratory of Child Neurodevelopment and Cognitive Disorders Chongqing China.
Pediatr Discov. 2024 Aug 2;2(3):e2500. doi: 10.1002/pdi3.2500. eCollection 2024 Sep.
The involvement of astrocytic connexin 43 (Cx43) in epileptogenesis has been extensively studied through various approaches, yet the underlying mechanism remains enigmatic. In this study, we explored whether astrocytic Cx43 forms hemichannels (HCs) that contribute to seizure progression in temporal lobe epilepsy (TLE) in mice. We focused on how these HCs influence the permeability of the blood-brain barrier (BBB), a crucial factor in the pathophysiology of epilepsy. Immunofluorescence staining and western blot analysis were employed to assess Cx43 expression in kainic acid-induced TLE mice, while BBB permeability was evaluated in TLE mice and those treated with TAT-Gap19 (an astrocytic Cx43 HC inhibitor) using Evans Blue permeation, serum S100β protein quantification, ZO-1 expression, and albumin extravasation into brain parenchyma via western blotting. Furthermore, seizure burden was monitored continuously using telemetric electroencephalography (EEG) and video monitoring in epileptic and TAT-Gap19-treated mice. Results demonstrated a significant increase in Cx43 content in hippocampal tissue in the TLE group, with a pronounced expression around blood vessels. TAT-GAP19 treatment alleviated EEG seizures and BBB permeability in TLE mice. These findings suggest that astrocytic Cx43 HCs in the hippocampus play a crucial role in epileptogenesis and seizure progression by regulating BBB permeability. Targeting Cx43-formed HCs distributed around the neurovascular unit may offer a novel therapeutic approach for epilepsy.
通过各种方法,人们对星形胶质细胞连接蛋白43(Cx43)在癫痫发生中的作用进行了广泛研究,但其潜在机制仍然不明。在本研究中,我们探讨了星形胶质细胞Cx43是否形成半通道(HCs),这些半通道有助于小鼠颞叶癫痫(TLE)的癫痫发作进展。我们重点研究了这些半通道如何影响血脑屏障(BBB)的通透性,而血脑屏障通透性是癫痫病理生理学中的一个关键因素。采用免疫荧光染色和蛋白质印迹分析来评估海藻酸诱导的TLE小鼠中Cx43的表达,同时使用伊文思蓝渗透、血清S100β蛋白定量、ZO-1表达以及通过蛋白质印迹法检测白蛋白渗入脑实质的情况,来评估TLE小鼠和用TAT-Gap19(一种星形胶质细胞Cx43 HC抑制剂)处理的小鼠的血脑屏障通透性。此外,通过遥测脑电图(EEG)和视频监测,持续监测癫痫小鼠和TAT-Gap19处理小鼠的癫痫发作负担。结果表明,TLE组海马组织中Cx43含量显著增加,血管周围有明显表达。TAT-GAP19治疗减轻了TLE小鼠的EEG癫痫发作和血脑屏障通透性。这些发现表明,海马中的星形胶质细胞Cx43 HCs通过调节血脑屏障通透性在癫痫发生和癫痫发作进展中起关键作用。针对分布在神经血管单元周围的由Cx43形成的HCs可能为癫痫提供一种新的治疗方法。