S1PR5 activation or overexpression enhances barrier function of mouse brain microvascular endothelial cells against OGD/R injury by modulating oxidative stress.
10.12122/j.issn.1673-4254.2025.07.11
- Author:
Jingxian WANG
1
;
Zijing REN
1
;
Peiyang ZHOU
1
Author Information
1. Department of Neurology, Xiangyang First People's Hospital, Hubei University of Medicine, Xiangyang 441000, China.
- Publication Type:Journal Article
- Keywords:
blood-brain barrier dysfunction;
oxidative stress;
oxygen-glucose deprivation and reoxygenation;
sphingosine 1-phosphate receptor 5
- MeSH:
Animals;
Mice;
Oxidative Stress;
Endothelial Cells/cytology*;
Brain/blood supply*;
Reactive Oxygen Species/metabolism*;
Receptors, Lysosphingolipid/metabolism*;
Sphingosine-1-Phosphate Receptors;
Blood-Brain Barrier/metabolism*;
Glucose;
Cell Line;
Oxygen/metabolism*;
NF-E2-Related Factor 2/metabolism*
- From:
Journal of Southern Medical University
2025;45(7):1451-1459
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES: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.
CONCLUSIONS: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.
