Abstract
The dysfunction of blood-brain barrier (BBB) plays a pivotal role in brain injury and subsequent neurological deficits of ischemic stroke. The current study aimed to examine the potential correlation between p53 inhibition and the neuroprotective effect of on the BBB. Rat middle cerebral artery occlusion and reperfusion model (MCAO/R) and oxygen-glucose deprivation/re-oxygenation model (OGD/R) were employed to simulate cerebral ischemia-reperfusion (CI/R) injury occurrence in vivo and in vitro. mNSS and TTC staining were applied to evaluate neurological deficits and brain infarct volumes. Evans blue (EB) staining was carried out to examine the permeability of BBB. RT-qPCR and Western blot to examine the mRNA and protein levels. Cell viabilities were detected by CCK-8. Flow cytometry and ELISA assay were employed to examine apoptosis and neuroinflammation levels. TEER value and sodium fluorescein were carried out to explore the permeability of HBMEC cells. PFT-α inhibited P53 and promoted the expression of β-catenin and cyclin D1, which were reversed by DKK1. PFT-α inhibited neurological deficits, brain infarct volume, neuroinflammation, apoptosis, and BBB integrity than the MCAO/R rats; however, this inhibition was reversed by DKK1. PFT-α promoted OGD/R-induced cell viability in NSCs, and suppressed inflammation and apoptosis, but DKK1 weakened the effect of PFT-α. PFT-α increased OGD/R-induced TEER values in cerebrovascular endothelial cells, inhibited sodium fluorescein permeability, and increased the mRNA levels of tight junction protein, but they were all attenuated by DKK1. PFT-α protects the BBB after acute ischemic stroke via the Wnt/β-catenin pathway, which in turn improves neurological function.
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This study was funded by the Natural Science Foundation of Shandong Province (No. ZR2020QH104) and Science and Technology Program of Binzhou Medical University (No. BY2018KJ06).
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HTZ, DYD, and XNG conceptualized and designed the study. XLT, YQX, and BW collected, organized, and drafted the information. All authors analyzed the data. HTZ, DYD, and XNG wrote the manuscript. SJY, PFL, and ZFL performed manuscript revision. All the authors have read and approved the manuscript.
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Animal care was conducted in compliance with the Guidelines for Care and Use of Laboratory Animals issued by the Ministry of Science and Technology of China, and the experimental protocols were passed by the Binzhou Medical University Hospital Animals Care and Use Committee. Every effort was made to reduce the number of animals as well as their suffering.
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ESM 1
Supplementary Figure 1 Diagram showing the experimental design of this study about the MCAO/R+PFT-α+DKK1 group, including the timing and number of rats for MACO/R, DKK1 treatment, PFT-α treatment, neurological deficit assessment, cerebral infarct volume, brain water content assessment, WB, RT-qPCR, ELISA assay. (PNG 397 kb)
ESM 2
Supplementary Figure 2 Diagram showing the experimental design of this study about the OGD/R+PFT-α+DKK1 group. A. The timing for detecting the effects of OGD/R, DKK1 and PFT-α on NSCs viability, apoptosis, inflammation and related signaling pathway proteins. B. The timing for detecting the effects of OGD/R, DKK1 and PFT-α on HBMECs permeability. (PNG 299 kb)
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Zhang, H., Du, D., Gao, X. et al. PFT-α protects the blood-brain barrier through the Wnt/β-catenin pathway after acute ischemic stroke. Funct Integr Genomics 23, 314 (2023). https://doi.org/10.1007/s10142-023-01237-3
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DOI: https://doi.org/10.1007/s10142-023-01237-3