The Combinational Treatment of Antioxidants Protects Brain Endothelial Cells Against Oxygen-Glucose Deprivation Followed by Reperfusion Induced Injury

Antioxidants including dehydroascorbic acid (DHA), glutathione (GSH) and melatonin have cellular protective effects on murine brain endothelial (bEnd.3) cells after oxidative stress and ischemic injury. Up to date, there are no studies that the combinational treatment of these antioxidants boosts the cellular protective effect on bEnd.3 cells. In this study, we investigated the protective effect of antioxidant combinational treatment on bEnd.3 cells after oxygen-glucose deprivation followed by reperfusion (OGD/R) induced injury. The effect of antioxidants combinational treatment on the bEnd.3 cells was examined by cell viability test, western blot analysis, immunocytochemistry. The combinational treatment of DHA (1 mM), GSH (1 mM) protects bEnd.3 cells after OGD/R induced injury and also melatonin (10 μM) treatment boosts the protective effect in combinational treatment of DHA and GSH on bEnd.3 cells after OGD/R induced injury. Western blot analysis suggests that the combinational treatment of DHA, GSH and melatonin attenuated the expression of vascular endothelial growth factor (VEGF), phosphorylation of ERK compared with each single treatment after OGD/R induced injury. Claudin 5, hypoxia inducible factor-1α (HIF-1α) and VEGF immunostaining data suggests the preventive effect of antioxidants combinational treatment. In conclusion, the combinational treatment of antioxidants elevates the cellular protective effect on bEnd.3cells against OGD/R induced injury.prohibitio