The expression of signaling kinases and specific junctional proteins (VE-cadherin, occludin, claudin, JAM-1) and transport proteins (MDR-1) were measured in rat’s brain endothelial cells cultured in vitro and exposed to simulated ischaemic conditions (OGD, oxygen-glucose deprivation). The effects of ebselen, Nacetylcysteine and glutamic acid were evaluated either in normoxic or simulated ischaemic conditions. Endothelial cells isolated from the fragments of rat brain microvessels seeded on the cell culture plates were cultured at 370 C in Dulbecco’s modified Eagle’s medium containing 20% fetal bovine serum, antibiotics and bFGF. OGD was obtained by incubation of cells in humidified atmosphere (3%O2, 92% N2, 5% CO2) in culture medium deprived of glucose and serum. Confluent cultures of endothelial cells were exposed to: 5000µM glutamic acid, 200µM N-acetylcysteine or 20µM ebselen. Intracellular signaling kinases (Akt, ERK1/2) and junctional and transporter proteins (VE-cadherin, occludin, claudin, JAM-1 and MDR-1) were detected using Western-blot. Ischemic conditions exerted negative effects on tight junctions of brain endothelial cells. ERK kinases were involved in the transduction of signals induced by ischemic conditions or by glutamic acid. Glutamic acid receptors were involved either in activation or inhibition of this pathway. In simulated ischaemic conditions ebselen as well as ebselen in combination with N-acetylocysteine exerted negative effects on intercellular junctions between brain endothelial cells. Antiooxidant compounds: N-acetylocysteine and ebselen affected MDR-1 level in brain endothelial cells. Supported by MNiSW grant 2P05A01530.
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