Activation of EphA4 induced by EphrinA1 exacerbates disruption of the blood-brain barrier following cerebral ischemia-reperfusion via the Rho/ROCK signaling pathway

Vascular dementia (VD) is a syndrome characterized by progressive cognitive decline. According to previous studies, stroke is considered to be a risk factor for VD. The disruption of the blood-brain barrier (BBB) is pivotal to the pathology of stroke, as it contributes to post-stroke inflammation and edema. It has been reported that the Eph/Ephrin signaling pathway serves an important role in central nervous system injury. However, the role of EphrinA1/EphA4 signaling in BBB damage following ischemic stroke has not yet been reported. Oxygen-glucose deprivation/reperfusion was performed to detect changes in EphrinA1 and EphA4 expression in human brain microvascular endothelial cells (HBMECs). Male mice were randomly divided into four groups [Sham, ischemia-reperfusion (I/R), I/R+EphrinA1 and I/R+EphA4] to observe the role of EphrinA1 and EphA4 under I/R conditions in vivo. The results of the present study revealed that the expression of EphrinA1 and EphA4 was significantly increased following I/R in vitro and in vivo. The administration of soluble ligand EphrinA1 enhanced CD68(+) cell accumulation, brain edema and dysfunction of the BBB, with lower expression levels of zonula occludens-1 (ZO-1) and Claudin-5. In addition, EphrinA1-treated mice had a higher level of caspase-3 and a lower level of phosphorylated-protein kinase B. However, the effects of EphrinA1 were abolished by EphA4-Fc, an inhibitor of EphA4. These results suggested that EphrinA1 exerted its effects on I/R injury via the activated EphA4 receptor. In addition, EphrinA1 decreased ZO-1 and Claudin-5 expression through the Rho/Rho associated kinase (ROCK) signaling pathway, which was attenuated by the pharmacological inhibition of Rho (C3 transferase) or ROCK (Y-27632). In conclusion, the present study provides evidence that the activation of EphA4 induced by EphrinA1 contributes to BBB damage following ischemic stroke through the Rho/ROCK signaling pathway, which highlights a potential therapeutic strategy for ischemic stroke and may help the development of preventative interventions for VD.