血脑屏障(BBB)破坏是缺血性卒中的主要病理生理事件。脑微血管内皮细胞对于维持中枢神经系统和外周系统之间的稳态至关重要。白藜芦醇预防缺血性中风。3,3\',4,5'-四甲氧基-反式-二苯乙烯(3,3',4,5\'-TMS)和3,4\',5-三甲氧基-反式-二苯乙烯(3,4',5-TMS)是添加甲氧基的白藜芦醇衍生物,表现出更好的药代动力学性能。我们旨在探讨其保护作用和潜在机制。将氧-葡萄糖剥夺(OGD)模型应用于bEnd.3细胞系,小鼠脑微血管内皮模拟缺血。用3,3\'预处理细胞,4,5'-TMS或3,4',5-TMS(1和5μM,24h),然后进行2hOGD损伤。细胞活力,促炎细胞因子和活性氧(ROS)的水平,通过分子检测和荧光染色检测蛋白质表达。OGD损伤引发细胞死亡,炎症反应,ROS产生和核因子-κB(NF-κB)信号通路。这些损伤被两个二苯乙烯显著减弱,3,3\',4,5\'-TMS和3,4\',5-TMS.它们还通过上调紧密连接蛋白的表达来减轻内皮屏障损伤。此外,3,3\',4,5\'-TMS和3,4\',5-TMS激活5'一磷酸腺苷活化蛋白激酶(AMPK)和内皮型一氧化氮合酶(eNOS)。总的来说,3,3\',4,5\'-TMS和3,4\',5-TMS通过抑制细胞死亡发挥对OGD损伤的保护作用,炎症反应,氧化应激,以及bEnd.3细胞上的BBB破坏。
Blood-brain barrier (BBB) disruption is a major pathophysiological event of ischemic stroke. Brain microvascular endothelial cells are critical to maintain homeostasis between central nervous system and periphery. Resveratrol protects against ischemic stroke. 3,3\',4,5\'-tetramethoxy-trans-stilbene (3,3\',4,5\'-TMS) and 3,4\',5-trimethoxy-trans-stilbene (3,4\',5-TMS) are resveratrol derivatives with addition of methoxy groups, showing better pharmacokinetic performance. We aimed to explore their protective effects and underlying mechanisms. Oxygen-glucose deprivation (OGD) model was applied in bEnd.3 cell line, mouse brain microvascular endothelium to mimic ischemia. The cells were pre-treated with 3,3\',4,5\'-TMS or 3,4\',5-TMS (1 and 5 μM, 24 h) and then subjected to 2-h OGD injury. Cell viability, levels of proinflammatory cytokines and reactive oxygen species (ROS), and protein expressions were measured by molecular assays and fluorescence staining. OGD injury triggered cell death, inflammatory responses, ROS production and nuclear factor-kappa B (NF-κB) signalling pathway. These impairments were remarkably attenuated by the two stilbenes, 3,3\',4,5\'-TMS and 3,4\',5-TMS. They also alleviated endothelial barrier injuries through upregulating the expression of tight junction proteins. Moreover, 3,3\',4,5\'-TMS and 3,4\',5-TMS activated 5\' adenosine monophosphate-activated protein kinase (AMPK) and endothelial nitric oxide synthase (eNOS). Overall, 3,3\',4,5\'-TMS and 3,4\',5-TMS exert protective effects against OGD damage through suppressing cell death, inflammatory responses, oxidative stress, as well as BBB disruption on bEnd.3 cells.