Sleep loss disrupts pericyte-brain endothelial cell interactions impairing blood-brain barrier function
Sleep loss disrupts pericyte-brain endothelial cell interactions impairing blood-brain barrier function
复制标题
DOI：
10.1016/j.bbi.2020.05.077
复制
发表时间：
2020-10-01
影响因子：
15.1
通讯作者：
Gomez-Gonzalez, Beatriz
中科院分区：
文献类型：
--
作者：
Medina-Flores, Fernanda;Hurtado-Alvarado, Gabriela;Gomez-Gonzalez, Beatriz
大鼠睡眠不足会破坏内皮间紧密连接，从而增加血脑屏障对循环分子的通透性。尽管描述了脑微血管的超微结构，并且有证据表明睡眠受限的大鼠中周细胞与毛细血管壁明显脱离，但睡眠不足对周细胞的影响尚不清楚。在这里，我们利用雄性 Wistar 大鼠描述了睡眠不足后周细胞和脑内皮细胞之间的相互作用。动物每天限制睡眠 20 小时，然后恢复 4 小时睡眠，持续 10 天。在睡眠限制结束时，从大脑皮层和海马体中分离出脑微血管 (MV)，并进行蛋白质印迹和免疫细胞化学处理，以评估周细胞-内皮细胞相互作用的标记物（连接蛋白 43、PDGFR-β）、紧密连接蛋白和促炎介质蛋白（MMP9、A(2A) 腺苷受体、CD73、NF kappa B）。睡眠限制降低了 MV 中 PDGFR-β 和连接蛋白 43 的表达；此外，扫描电子显微镜照片显示周细胞与毛细血管壁分离，但没有发生细胞凋亡（如活性 caspase-3 表达减少所示）。在体内渗透性测定中，睡眠限制还降低了 MV 中紧密连接蛋白的表达，并增加了 BBB 对低分子量和高分子量示踪剂的渗透性。这些改变似乎取决于低度炎症状态，睡眠受限大鼠脑内皮细胞中磷酸化 NF kappa B 和 A2A 腺苷受体表达的增加反映了这一点。我们的数据表明，睡眠限制会改变周细胞-脑内皮细胞的相互作用。这一证据对于了解睡眠在调节血脑屏障功能中的作用至关重要。
Sleep loss in the rat increases blood-brain barrier permeability to circulating molecules by disrupting inter-endothelial tight junctions. Despite the description of the ultrastructure of cerebral microvessels and the evidence of an apparent pericyte detachment from capillary wall in sleep restricted rats the effect of sleep loss on pericytes is unknown. Here we characterized the interactions between pericytes and brain endothelial cells after sleep loss using male Wistar rats. Animals were sleep-restricted 20 h daily with 4 h sleep recovery for 10 days. At the end of the sleep restriction, brain microvessels (MVs) were isolated from cerebral cortex and hippocampus and processed for Western blot and immunocytochemistry to evaluate markers of pericyte-endothelial cell interaction (connexin 43, PDGFR-beta), tight junction proteins, and proinflammatory mediator proteins (MMP9, A(2A) adenosine receptor, CD73, NF kappa B). Sleep restriction reduced PDGFR-beta and connexin 43 expression in MVs; in addition, scanning electron microscopy micrographs showed that pericytes were detached from capillary walls, but did not undergo apoptosis (as depicted by a reduced active caspase-3 expression). Sleep restriction also decreased tight junction protein expression in MVs and increased BBB permeability to low- and high-molecular weight tracers in in vivo permeability assays. Those alterations seemed to depend on a low-grade inflammatory status as reflected by the increased expression of phosphorylated NF kappa B and A2A adenosine receptor in brain endothelial cells from the sleep-restricted rats. Our data show that pericyte-brain endothelial cell interaction is altered by sleep restriction; this evidence is essential to understand the role of sleep in regulating blood-brain barrier function.