GSK-3β inhibitor TWS119 attenuates rtPA-induced hemorrhagic transformation and activates the Wnt/β-catenin signaling pathway after acute ischemic stroke in rats

Abstract

Hemorrhagic transformation (HT) is a devastating complication for patients with acute ischemic stroke who are treated with tissue plasminogen activator (tPA). It is associated with high morbidity and mortality, but no effective treatments are currently available to reduce HT risk. Therefore, methods to prevent HT are urgently needed. In this study, we used TWS119, an inhibitor of glycogen synthase kinase 3β (GSK-3β), to evaluate the role of the Wnt/β-catenin signaling pathway in recombinant tPA (rtPA)-induced HT. Sprague–Dawley rats were subjected to a middle cerebral artery occlusion (MCAO) model of ischemic stroke and then were administered rtPA, rtPA combined with TWS119, or vehicle at 4 h. The animals were sacrificed 24 h after infarct induction. Rats treated with rtPA showed evident HT, had more severe neurologic deficit, brain edema, and blood–brain barrier breakdown, and had larger infarction volume than did the vehicle group. Rats treated with TWS119 had significantly improved outcomes compared with those of rats treated with rtPA alone. In addition, Western blot analysis showed that TWS119 increased the protein expression of β-catenin, claudin-3, and ZO-1 while suppressing the expression of GSK-3β. These results suggest that TWS119 reduces rtPA-induced HT and attenuates blood–brain barrier disruption, possibly through activation of the Wnt/β-catenin signaling pathway. This study provides a potential therapeutic strategy to prevent tPA-induced HT after acute ischemic stroke.

Inhibition of tPA-induced hemorrhagic transformation involves adenosine A2b receptor activation after cerebral ischemia

“…A sample size calculation based on our previous study (Wang et al, 2015e) and pilot data indicated that 6 rats/group would provide at least 80% power for detecting a 20% decrease in hemoglobin content at α = 0.05 (2-sided). Rats were randomly allocated into study groups by using the website Randomization.com (Cheng et al, 2016).…”

“…Neurologic deficits were assessed at 24 h post-ischemia with a neurologic deficit grading system as described previously (Wang et al, 2015e). Each animal was scored on a scale of 0 to 5.…”

Gpr124 is essential for blood–brain barrier integrity in central nervous system disease

“…During homeostasis, endothelial Fzd4 deletion produces BBB compromise in adult mice 12 , whereas conditional deletion of endothelial Ctnnb1 in adult mice induces BBB breakdown and brain petechial hemorrhage, either through transcriptional activation or the adherens-junction functions of β-catenin 24 . During pathologic conditions, Wnt–β-catenin signaling in unfractionated brain tissue is correlated with tissue damage and hemorrhagic transformation after ischemic stroke 25,26 , and endothelial Wnt–β-catenin signaling is functionally implicated in the regulation of vascular integrity in glioblastoma 27 .…”

Wnt Pathway: An Emerging Player in Vascular and Traumatic Mediated Brain Injuries

“…These results suggest that activation of the canonical Wnt pathway through Dkk1 inhibition and GSK3β deactivation jointly contribute to the neuroprotective effects of HPC against ischemic injury (Zhan et al, 2019). Moreover, the acute administration of 4,6-disubstituted pyrrolopyrimidine (TWS119), a specific inhibitor of GSK3β, has been shown to provide neuroprotection by decreasing the infarct volume and reducing BBB permeability after cerebral ischemia in rodents (Wang W. et al, 2016). These effects were associated to canonical Wnt signaling pathway activation, which increased the expression of tight junction proteins claudin-3 and ZO1 (Wang W. et al, 2016).…”