Background Excitotoxicity is a central pathological pathway in many neurological diseases with blood–brain barrier (BBB) dysfunction. Kainate, an exogenous excitotoxin, induces epilepsy and BBB damage in animal models, but the direct effect of kainate on brain endothelial cells has not been studied in detail. Our aim was to examine the direct effects of kainate on cultured cells of the BBB and to test three anti-inflammatory and antioxidant drugs used in clinical practice, simvastatin, edaravone and dexamethasone, to protect against kainate-induced changes. Methods Primary rat brain endothelial cell, pericyte and astroglia cultures were used to study cell viability by impedance measurement. BBB permeability was measured on a model made from the co-culture of the three cell types. The production of nitrogen monoxide and reactive oxygen species was followed by fluorescent probes. The mRNA expression of kainate receptors and nitric oxide synthases were studied by PCR. Results Kainate damaged brain endothelial cells and made the immunostaining of junctional proteins claudin-5 and zonula occludens-1 discontinuous at the cell border indicating the opening of the barrier. The permeability of the BBB model for marker molecules fluorescein and albumin and the production of nitric oxide in brain endothelial cells were increased by kainate. Simvastatin, edaravone and dexamethasone protected against the reduced cell viability, increased permeability and the morphological changes in cellular junctions caused by kainate. Dexamethasone attenuated the elevated nitric oxide production and decreased the inducible nitric oxide synthase (NOS2/iNOS) mRNA expression increased by kainate treatment. Conclusion Kainate directly damaged cultured brain endothelial cells. Simvastatin, edaravone and dexamethasone protected the BBB model against kainate-induced changes. Our results confirmed the potential clinical usefulness of these drugs to attenuate BBB damage.

中文翻译：

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辛伐他汀、依达拉奉和地塞米松防止红藻氨酸诱导的脑内皮细胞损伤

背景兴奋性毒性是许多具有血脑屏障 (BBB) 功能障碍的神经系统疾病的主要病理途径。海人酸是一种外源性兴奋性毒素，在动物模型中诱导癫痫和 BBB 损伤，但海人酸对脑内皮细胞的直接影响尚未详细研究。我们的目的是检查红藻氨酸对 BBB 培养细胞的直接影响，并测试临床实践中使用的三种抗炎和抗氧化药物，辛伐他汀、依达拉奉和地塞米松，以防止红藻氨酸引起的变化。方法采用原代大鼠脑内皮细胞、周细胞和星形胶质细胞培养物，通过阻抗测量研究细胞活力。在由三种细胞类型的共培养物制成的模型上测量 BBB 通透性。一氧化氮和活性氧的产生之后是荧光探针。通过PCR研究红藻氨酸受体和一氧化氮合酶的mRNA表达。结果海人酸损伤脑内皮细胞并使连接蛋白claudin-5和封闭小带1的免疫染色在细胞边界处不连续，表明屏障的开放。红藻氨酸增加了 BBB 模型对标记分子荧光素和白蛋白的渗透性以及脑内皮细胞中一氧化氮的产生。辛伐他汀、依达拉奉和地塞米松可防止由红藻氨酸引起的细胞活力降低、通透性增加和细胞连接处的形态变化。地塞米松减弱了升高的一氧化氮产生并降低了由红藻氨酸处理增加的诱导型一氧化氮合酶 (NOS2/iNOS) mRNA 表达。结论海人酸直接损伤培养的脑内皮细胞。辛伐他汀、依达拉奉和地塞米松保护 BBB 模型免受红藻氨酸诱导的变化。我们的结果证实了这些药物在减轻 BBB 损伤方面的潜在临床用途。