Brain endothelial permeability plays a crucial role in blood-brain barrier (BBB), but the permeability enhancement in cerebral ischemia reperfusion (I/R). Vitexin has certain neuroprotective effects, but the effect brain endothelial permeability in I/R injury was unknown. In this study, the effects of Vitexin on endothelial permeability and the underlying mechanisms in human brain microvascular endothelial cells (HBMEc) I/R injury model were investigated. Cell viability, lactate dehydrogenase (LDH), inflammation and apoptosis were detected. The effects of Vitexin on BBB integrity tight junction, matrix metalloproteinases (MMP) were also investigated. The mechanism was confirmed by PI3K inhibitor and NOS inhibitor in normal or eNOS siRNA transfection HBMEc. Vitexin significantly reduced LDH, Caspase 3 level, alleviated inflammation, also could maintain BBB integrity, increased tight junction proteins expression and inhibited MMP. The mechanism is related to reduction of intracellular NO and ONOO-, regulated eNOS, iNOS activity. Vitexin significantly preserved eNOS phosphorylation in response to the activated Akt. Moreover, combined with PI3K inhibitor or low dosage of NOS inhibitor, totally abolished Vitexin-induced eNOS phosphorylation, the protected effect was also attenuated, but still significantly between model cells. However, combined with high dosage NOS inhibitor which both inhibited the eNOS phosphorylation and iNOS, the protected effect of Vitexin was abrogated. In addition, eNOS silencing cells were used to further clarify the regulatory role of Vitexin on iNOS. Our findings showed that Vitexin could play a protective role in I/R-induced brain endothelial permeability by simultaneously increase eNOS phosphorylation and inhibit iNOS.
Keywords: Blood–brain barrier; Cerebral ischemia reperfusion; Vitexin; eNOS; iNOS.
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