He Chongxin, Bao Yong, Xu Yong, Cheng Jingjing, Hu Xinxin
Department of Neurosurgery, Hefei Third People's Hospital, Hefei Third Clinical College of Anhui Medical University, Hefei, Anhui, China.
Department of Neurology, Hefei Third People's Hospital, Hefei Third Clinical College of Anhui Medical University, Hefei, Anhui, China.
Biomol Biomed. 2025 Jan 30;25(3):553-562. doi: 10.17305/bb.2024.11366.
Ischemic stroke often results in high mortality and significant disability. Current research primarily focuses on understanding neuroinflammation and cell death following a stroke to identify novel therapeutic targets. This study investigates the endothelial cell-specific role of Thioredoxin interacting protein (TXNIP) in ischemic stroke and its underlying molecular mechanisms both in vitro and in vivo. By targeting endothelial cells, we aim to determine how TXNIP knockdown promotes neuroprotection, enhances angiogenesis, and reduces inflammation post-stroke. In vitro, an oxygen-glucose deprivation (OGD) model using bEnd.3 cells simulated ischemic conditions. Cellular injury was evaluated through cell proliferation and angiogenesis assays, while dual immunofluorescence staining assessed ZO-1 and CD31 expression. Western blotting measured protein levels of TXNIP, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), ASC, pro-caspase-1, and interleukin-1β (IL-1β). In vivo, a middle cerebral artery occlusion (MCAO) mouse model was employed to mimic ischemic stroke. Brain injury was evaluated using triphenyl tetrazolium chloride (TTC) and Nissl staining, and molecular changes in injury markers were assessed via Western blot analysis. In vitro, TXNIP knockdown promoted cell proliferation and angiogenesis, reduced inflammation, and decreased ZO-1 and CD31 fluorescence intensity. TXNIP knockdown also reversed OGD-induced upregulation of TXNIP, NLRP3, ASC, pro-caspase-1, and IL-1β. In vivo, TXNIP knockdown improved neurological recovery, reflected by lower Longa scores, increased Nissl body presence, and reduced infarct size. These findings suggest that TXNIP knockdown mitigates inflammation, enhances angiogenesis, and reduces cerebral damage following ischemic stroke. This provides valuable insights into potential endothelial cell-specific therapeutic strategies for stroke treatment.
缺血性中风常导致高死亡率和严重残疾。当前研究主要集中在了解中风后的神经炎症和细胞死亡，以确定新的治疗靶点。本研究在体外和体内研究硫氧还蛋白相互作用蛋白（TXNIP）在内皮细胞特异性在缺血性中风中的作用及其潜在分子机制。通过靶向内皮细胞，我们旨在确定TXNIP基因敲低如何促进神经保护、增强血管生成并减少中风后的炎症。在体外，使用bEnd.3细胞的氧-葡萄糖剥夺（OGD）模型模拟缺血条件。通过细胞增殖和血管生成测定评估细胞损伤，同时采用双重免疫荧光染色评估ZO-1和CD31表达。蛋白质印迹法检测TXNIP、核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）、凋亡相关斑点样蛋白（ASC）、前半胱天冬酶-1和白细胞介素-1β（IL-1β）的蛋白水平。在体内，采用大脑中动脉闭塞（MCAO）小鼠模型模拟缺血性中风。使用氯化三苯基四氮唑（TTC）和尼氏染色评估脑损伤，并通过蛋白质印迹分析评估损伤标志物的分子变化。在体外，TXNIP基因敲低促进细胞增殖和血管生成，减轻炎症，并降低ZO-1和CD31荧光强度。TXNIP基因敲低还逆转了OGD诱导的TXNIP、NLRP3、ASC、前半胱天冬酶-1和IL-1β的上调。在体内，TXNIP基因敲低改善了神经功能恢复，表现为较低的Longa评分、尼氏体数量增加和梗死面积减小。这些发现表明，TXNIP基因敲低可减轻缺血性中风后的炎症，增强血管生成并减少脑损伤。这为中风治疗中潜在的内皮细胞特异性治疗策略提供了有价值的见解。
