OGD-pretreated astrocyte-derived extracellular vesicles attenuates intracerebral hemorrhage (ICH)-induced BBB disruption through miR-27a-3p /ARHGAP25 /Wnt/β- catenin axis

Abstract

Abstract Background Breakdown of blood brain barrier (BBB) is one of the key mechanisms of brain injury following intracerebral hemorrhage (ICH). Astrocytes interact with endothelial and contribute to the regulation of neurovascular coupling via paracrine signaling factors. Increasing number of studies now point to astrocyte-derived extracellular vesicles (ADEVs) as an important way of intercellular information communication. However, the role of ADEVs in the regulation of the BBB integrity after ICH remains unclear. Methods EVs were obtained from astrocytes with or without oxygen and glucose deprivation (OGD) pre-stimulation and the role of ADEVs in ICH was investigated ICH rat model and ICH cell model. The potential regulatory effect of ADEVs on endothelial barrier integrity was identified by TEER, western blot and immunofluorescence in vitro. In vivo, functional evaluation, Evans-blue leakage and tight junction proteins (TJPs) expression were analyzed. MiRNA sequencing revealed that microRNA-27a-3p (miR-27a-3p) was differentially expressed miRNA in the extracellular vesicles (EVs) from OGD-pretreated astrocytes, compared with normal control. The regulatory mechanism of miR-27a-3p was assessed using Luciferase assay, RT-PCR, western blot and immunofluorescence. Results OGD-activated astrocytes reduced hemin-induced endothelial hyper-permeability through secreting EVs. OGD-activated ADEVs alleviated BBB dysfunction after ICH in vivo and in vitro. MicroRNA microarray analysis indicated that miR-27a-3p is a major component that was highly expressed miRNA in OGD pretreated-ADEVs. OGD-ADEVs mitigated BBB injury through transferring miR-27a-3p into bEnd.3 cells and regulating ARHGAP25/Wnt/β-catenin pathway. Conclusion Taken together, these findings firstly reveal that EVs from OGD pre-stimulated astrocytes improve functional recovery through counteracting BBB injury by regulating endothelial ARHGAP25/Wnt/β-catenin axis after ICH. EVs released from hypoxia astrocytes might be a novel strategy for the targeting treatment of ICH.