S1PR5 activation or overexpression enhances barrier function of mouse brain microvascular endothelial cells against OGD/R injury by modulating oxidative stress
WANG Jingxian
REN Zijing
ZHOU Peiyang
Abstract：Objective To investigate the role of sphingosine-1-phosphate receptor 5(S1PR5)in modulating barrier function of mouse brain microvascular endothelial cells with oxygen-glucose deprivation and reoxygenation(OGD/R).Methods Mouse brain microvascular endothelial cells(bEnd.3)were exposed to OGD/R to induce barrier dysfunction following treatment with S1PR5-specific agonist A971432 or lentivirus-mediated transfection with a S1PR5-specific siRNA,a S1PR5-overexpressing plasmid,or their respective negative control sequences.The changes in viability and endothelial barrier permeability of the treated cells were evaluated with CCK-8 assay and FITC-dextran permeability assay;the levels of intracellular reactive oxygen species(ROS)and localization and expression levels of the proteins related with barrier function and oxidative stress were detected using immunofluorescence staining,DCFH-DA probe and Western blotting.Results S1PR5 activation obviously enhanced viability of bEnd.3 cells exposed to OGD/R(P<0.0001).Both activation and overexpression of S1PR5 reduced FITC-dextran leakage,while S1PR5 knockdown significantly increased FITC-dextran leakage in the exposed bEnd.3 cells.Activation and overexpression of S1PR5 both increased the cellular expressions of the barrier proteins ZO-1 and occludin,while S1PR5 knockdown produced the opposite effect.In cells exposed to OGD/R,ROS production was significantly reduced by S1PR5 activation and overexpression but increased following S1PR5 knockdown.Overexpression of S1PR5 obviously increased the expressions of the antioxidant proteins Nrf2,HO-1 and SOD2 in the exposed cells.Conclusion S1PR5 activation and overexpression significantly improve cell viability and reduce permeability of a mouse brain microvascular endothelial cell model of OGD/R,the mechanism of which may involve the reduction in ROS production and upregulation of the antioxidant proteins.
Keywords：sphingosine 1-phosphate receptor 5blood-brain barrier dysfunctionoxidative stressoxygen-glucose deprivation and reoxygenation
Publication Date：2025-07-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：9( 1451-1459 )