南方医科大学学报2025，Vol.45Issue(7)：1451-1459,9.DOI:10.12122/j.issn.1673-4254.2025.07.11
S1PR5激动与过表达通过调控氧化应激增强脑微血管内皮细胞屏障功能抵抗氧糖剥夺/复氧复糖损伤
S1PR5 activation or overexpression enhances barrier function of mouse brain microvascular endothelial cells against OGD/R injury by modulating oxidative stress
摘要
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.关键词
鞘氨醇1-磷酸受体5/血脑屏障功能障碍/氧化应激/氧糖剥夺/复氧复糖损伤Key words
sphingosine 1-phosphate receptor 5/blood-brain barrier dysfunction/oxidative stress/oxygen-glucose deprivation and reoxygenation引用本文复制引用
王静娴,任自敬,周佩洋..S1PR5激动与过表达通过调控氧化应激增强脑微血管内皮细胞屏障功能抵抗氧糖剥夺/复氧复糖损伤[J].南方医科大学学报,2025,45(7):1451-1459,9.基金项目
湖北省自然科学基金(2021CFB567) （2021CFB567）
湖北医药学院研究生创新项目(YC2023048) （YC2023048）
