Simvastatin, edaravone and dexamethasone protect against kainate-induced brain endothelial cell damage

Abstract

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. Publisher's NoteSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Combination of Alanine and Glutathione as Targeting Ligands of Nanoparticles Enhances Cargo Delivery into the Cells of the Neurovascular Unit

“…Isolation of primary rat brain endothelial cells, pericytes and astrocytes were performed according to the method described in our previous studies [ 27 , 28 ]. After isolation, brain endothelial cells were seeded onto culture dishes (Corning Costar, New York, NY, USA) coated with collagen type IV (100 μg/mL) and fibronectin (25 μg/mL) and were cultured in DMEM/HAM’s F-12 (Gibco, Waltham, MA, USA) supplemented with 15% plasma-derived bovine serum (PDS, First Link, UK), 10 mM HEPES, 100 μg/mL heparin, 5 μg/mL insulin, 5 μg/mL transferrin, 5 ng/mL sodium selenite (ITS, Pan-Biotech, Aidenbach, Germany), 1 ng/mL basic fibroblast growth factor (bFGF, Roche, Basel, Switzerland) and 50 μg/mL gentamicin.…”

Ameliorative effects of ceftriaxone sodium combined with dexamethasone on infantile purulent meningitis and associated effects on brain‑derived neurotrophic factor levels

“…As a result, damage to the neurovascular system is greatly reduced. In addition, dexamethasone contributed to the stabilization of the vascular endothelial structure and was of great significance to the protection of neurons and blood vessels (29). This was speculated to be a reason for the higher BDNF levels observed in the combination group compared with the monotherapy group in the present study.…”

MAP Kinase Pathways in Brain Endothelial Cells and Crosstalk with Pericytes and Astrocytes Mediate Contrast-Induced Blood–Brain Barrier Disruption

“…The cell types used in the triple co-culture BBB model have been characterized in detail in our previous publications [ 15 , 18 , 20 ]. Brain endothelial cell cultures expressed endothelial characteristics [ 15 ] as well as BBB markers both at the gene and protein levels [ 15 , 21 ].…”

“…Brain endothelial cell cultures expressed endothelial characteristics [ 15 ] as well as BBB markers both at the gene and protein levels [ 15 , 21 ]. More than 90% of the cells in the astrocyte cultures stained positively for glial fibrillary acidic protein [ 15 , 18 , 20 ]. Pericyte cultures expressed NG-2 and alpha-smooth muscle actin [ 15 , 18 , 20 ].…”