Abstract：
Blood-brain barrier (BBB) damage significantly affects the prognosis of ischemic stroke patients. This project employed multi-omics analysis to identify key factors regulating BBB disruption during cerebral ischemia-reperfusion. An integrated analysis of three transcriptome sequencing datasets from mouse middle cerebral artery occlusion/reperfusion (MCAO/R) models identified eight downregulated genes in endothelial cells. Additionally, transcriptome analysis of BBB (cortex) and non-BBB (lung) endothelium of E13.5 mice revealed 2,102 upregulated genes potentially associated with BBB integrity. The eight downregulated genes were intersected with the 2,102 BBB-related genes and mapped using single-cell RNA sequencing data, revealing that solute carrier family 22 member 8 (Slc22a8) is specifically expressed in endothelial cells and pericytes and significantly decreases after MCAO/R. This finding was validated in the mouse MCAO/R model at both protein and mRNA levels in this study. External overexpression of Slc22a8 using a lentivirus carrying Tie2 improved Slc22a8 and tight junction protein levels and reduced BBB leakage after MCAO/R, accompanied by Wnt/β-catenin signaling activation. In conclusion, this study suggested that MCAO/R-induced downregulation of Slc22a8 expression may be a crucial mechanism underlying BBB disruption. Interventions that promote Slc22a8 expression or enhance its function hold promise for improving the prognosis of patients with cerebral ischemia.
摘要：
血脑屏障(BBB)损伤显著影响缺血性脑卒中患者的预后。该项目采用多组学分析来确定脑缺血再灌注过程中调节BBB破坏的关键因素。对来自小鼠大脑中动脉阻塞/再灌注(MCAO/R)模型的三个转录组测序数据集的综合分析鉴定了内皮细胞中的八个下调基因。此外,E13.5小鼠的BBB（皮质）和非BBB（肺）内皮的转录组分析显示，2,102个上调的基因可能与BBB完整性相关。8个下调基因与2,102个BBB相关基因相交，并使用单细胞RNA测序数据进行定位，揭示溶质载体家族22成员8(Slc22a8)在内皮细胞和周细胞中特异性表达，并在MCAO/R后显著降低。在该研究中,在小鼠MCAO/R模型中在蛋白质和mRNA水平上验证了该发现。使用携带Tie2的慢病毒对Slc22a8的外部过表达改善了Slc22a8和紧密连接蛋白水平，并减少了MCAO/R后的BBB渗漏，伴有Wnt/β-catenin信号激活。总之,这项研究表明，MCAO/R诱导的Slc22a8表达下调可能是BBB破坏的关键机制。促进Slc22a8表达或增强其功能的干预措施有望改善脑缺血患者的预后。
