Yan Bing Chun, Xu Pei, Gao Manman, Wang Jie, Jiang Dan, Zhu Xiaolu, Won Moo-Ho, Su Pei Qing
Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Department of Traditional Chinese and Western Medicine, Yangzhou University, Yangzhou, China.
Department of Integrated Traditional Chinese and Western Medicine, Medical College, Yangzhou University, Yangzhou, China.
Front Neurol. 2018 Sep 12;9:775. doi: 10.3389/fneur.2018.00775. eCollection 2018.
The kainic acid (KA)-induced epilepsy experimental model is widely used to study the mechanisms underlying this disorder. Recently, the blood-brain barrier (BBB) has become an innovative alternative treatment target for epilepsy patients. KA causes neuronal injury and BBB damage in this experimental epilepsy model but the mechanisms underlying epilepsy-related neuronal injury, autophagy, and BBB damage remain unclear. Therefore, the present study investigated the relationships among neuronal injury, the expressions of autophagy-related proteins, and changes in BBB-related proteins during the acute phase of epilepsy to further understand the mechanisms and pharmacotherapy of epilepsy. NeuN immunohistochemistry and Fluoro-Jade B (FJ-B) staining in the hippocampal CA3 region revealed that neuronal death induced by intraventricular injections of 10 μg/kg KA was greater than that induced by 3 μg/kg KA. In addition, there were transient increases in the levels of microtubule-associated protein light chain 3-II (LC3I/II) and Beclin-1, which are autophagy-related proteins involved in neuronal death, in this region 24 h after the administration of 10 μg/kg KA. There were also morphological changes in BBB-related cells such as astrocytes, endothelial cells (ECs), and tight junctions (TJs). More specifically, there was a significant increase in the activation of astrocytes 72 h after the administration of 10 μg/kg KA as well as continuous increases in the expressions of platelet endothelial cell adhesion molecule-1 (PECAM-1) and BBB-related TJ proteins (Zonula occludens-1 and Claudin-5) until 72 h after KA treatment. These results suggest that the overexpression of autophagy-related proteins and astrocytes and transient increases in the expressions of BBB-related TJ proteins may be closely related to autophagic neuronal injury. These findings provide a basis for the identification of novel therapeutic targets for patients with epilepsy.
海藻酸（KA）诱导的癫痫实验模型被广泛用于研究这种疾病的潜在机制。最近，血脑屏障（BBB）已成为癫痫患者一种创新的替代治疗靶点。在这个实验性癫痫模型中，KA会导致神经元损伤和血脑屏障破坏，但癫痫相关神经元损伤、自噬和血脑屏障破坏的潜在机制仍不清楚。因此，本研究调查了癫痫急性期神经元损伤、自噬相关蛋白表达和血脑屏障相关蛋白变化之间的关系，以进一步了解癫痫的机制和药物治疗。海马CA3区的NeuN免疫组织化学和荧光玉髓B（FJ - B）染色显示，脑室内注射10μg/kg KA诱导的神经元死亡比3μg/kg KA诱导的更严重。此外，在给予10μg/kg KA后24小时，该区域中参与神经元死亡的自噬相关蛋白微管相关蛋白轻链3 - II（LC3I/II）和Beclin - 1的水平出现短暂升高。血脑屏障相关细胞如星形胶质细胞、内皮细胞（ECs）和紧密连接（TJs）也有形态学变化。更具体地说，给予10μg/kg KA后72小时，星形胶质细胞的活化显著增加，并且血小板内皮细胞黏附分子-1（PECAM - 1）和血脑屏障相关TJ蛋白（闭合蛋白-1和Claudin - 5）的表达持续增加直至KA治疗后72小时。这些结果表明，自噬相关蛋白和星形胶质细胞的过度表达以及血脑屏障相关TJ蛋白表达的短暂增加可能与自噬性神经元损伤密切相关。这些发现为确定癫痫患者的新治疗靶点提供了依据。
