Duan Yu, Deng Yao, Tang Feng, Li Jian
Department of Neurosurgery, Huadong Hospital Affiliated to Fudan University, Jing’an, Shanghai 200040, China.
Aging (Albany NY). 2024 Mar 27;16(7):6135-6146. doi: 10.18632/aging.205692.
Dysfunction of tight junction proteins-associated damage to the blood-brain barrier (BBB) plays an important role in the pathogenesis of ischemic stroke. Lifibrate, an inhibitor of cholinephosphotransferase (CPT), has been used as an agent for serum lipid lowering. However, the protective effects of Lifibrate in ischemic stroke and the underlying mechanism have not been clearly elucidated. Here, we employed an mice model of MCAO and an OGD/R model . In the mice models, neurological deficit scores and infarct volume were assessed. Evans Blue solution was used to detect the BBB permeability. The TEER was examined to determine brain endothelial monolayer permeability. Here, we found that Lifibrate improved neurological dysfunction in stroke. Additionally, increased BBB permeability during stroke was significantly ameliorated by Lifibrate. Correspondingly, the reduced expression of the tight junction protein ZO-1 was restored by Lifibrate at both the mRNA and protein levels. Using an model, we found that Lifibrate ameliorated OGD/R-induced injury in human bEnd.3 brain microvascular endothelial cells by increasing cell viability but reducing the release of LDH. Importantly, Lifibrate suppressed the increase in endothelial monolayer permeability and the reduction in TEER induced by OGD/R via the rescue of ZO-1 expression. Mechanistically, Lifibrate blocked activation of the MLCK/ p-MLC signaling pathway in OGD/R-stimulated bEnd.3 cells. In contrast, overexpression of MLCK abolished the protective effects of Lifibrate in endothelial monolayer permeability, TEER, as well as the expression of ZO-1. Our results provide a basis for further investigation into the neuroprotective mechanism of Lifibrate during stroke.
紧密连接蛋白功能障碍相关的血脑屏障（BBB）损伤在缺血性中风的发病机制中起重要作用。利贝特，一种胆碱磷酸转移酶（CPT）抑制剂，已被用作降血脂药物。然而，利贝特在缺血性中风中的保护作用及其潜在机制尚未明确阐明。在此，我们采用了小鼠大脑中动脉闭塞（MCAO）模型和氧糖剥夺/复氧（OGD/R）模型。在小鼠模型中，评估神经功能缺损评分和梗死体积。使用伊文思蓝溶液检测血脑屏障通透性。检测跨上皮电阻（TEER）以确定脑内皮单层通透性。在此，我们发现利贝特改善了中风后的神经功能障碍。此外，利贝特显著改善了中风期间血脑屏障通透性的增加。相应地，利贝特在mRNA和蛋白质水平上恢复了紧密连接蛋白ZO-1表达的降低。使用OGD/R模型，我们发现利贝特通过提高细胞活力但减少乳酸脱氢酶（LDH）的释放，改善了OGD/R诱导的人bEnd.3脑微血管内皮细胞损伤。重要的是，利贝特通过挽救ZO-1表达，抑制了OGD/R诱导的内皮单层通透性增加和TEER降低。机制上，利贝特阻断了OGD/R刺激的bEnd.3细胞中肌球蛋白轻链激酶（MLCK）/磷酸化肌球蛋白轻链（p-MLC）信号通路的激活。相反，MLCK的过表达消除了利贝特在内皮单层通透性、TEER以及ZO-1表达方面的保护作用。我们的结果为进一步研究利贝特在中风期间的神经保护机制提供了依据。
