Effects of hypoxia on endothelial/pericytic co-culture model of the blood-brain barrier
Effects of hypoxia on endothelial/pericytic co-culture model of the blood-brain barrier
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DOI：
10.1016/j.regpep.2004.05.023
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发表时间：
2004-12-15
影响因子：
--
通讯作者：
Niwa, M
中科院分区：
文献类型：
--
作者：
Hayashi, K;Nakao, S;Niwa, M
血脑屏障（BBB）由内皮细胞、周细胞和星形胶质细胞足突组成。对于体外血脑屏障的研究大多数是在有或没有星形胶质细胞的内皮细胞上进行的。血脑屏障的缺氧损伤诱导血管源性脑水肿。我们已经产生了一个新的模型的血脑屏障与脑内皮细胞和周细胞，并检查了缺氧的影响，使用这个model.Brain微血管内皮细胞和周细胞分离从三周龄的雄性Wister大鼠使用酶和机械匀浆。三种型号（A：仅内皮单层，B：通过使用Transwell共培养系统培养这些细胞来制备具有周细胞的内皮单层（非接触条件，和C：接触条件），并将其暴露于缺氧条件。我们用跨内皮电阻（TEER）和伊文思蓝-白蛋白和荧光素钠的通透性来评价屏障功能，在内皮/细胞周接触模型中观察到最紧密的屏障。尽管在内皮单层和非接触共培养模型中缺氧诱导的屏障破坏，但在接触共培养模型中观察到最低限度的功能障碍。因此，认为周细胞通过分泌因子或缝隙连接作用于内皮，诱导内皮成熟，形成更紧密的屏障功能，从而支持抗缺氧损伤的功能。细胞间通讯可能是维持缺氧条件下血脑屏障功能和稳定的重要因素。(C)2004 Elsevier B. V.保留所有权利。
The blood-brain barrier (BBB) is composed of endothelial cells, pericytes and astrocytic foot processes. Most research for the in vitro BBB is performed endothelial cells with or without astrocytes. Hypoxia damage to the BBB induces vasogenic brain edema. We have generated a new model of the BBB with brain endothelial cells and pericytes and have examined the effects of hypoxia using this model.Brain microvascular endothelial cells and pericytes were isolated from three-week-old male Wister rats using enzyme and mechanical homogenization. Three models (A: only endothelial monolayer, B: endothelial monolayer with pericytes non-contact condition, and C: contact condition) were made by culturing these cells using Transwell co-culture system and were exposed to hypoxic condition. We evaluated barrier function with transendothelial electrical resistance (TEER) and permeability of Evans blue-albumin and sodium fluorescein.The tightest barrier was observed in the endothelial/pericytic contact model. Despite hypoxia-induced disruption of the barrier in endothelial monolayer and non-contact co-culture models, a minimum of dysfunction was seen in the contact co-culture model. Therefore, it is considered that pericytes effect on the endothelia by secreting factors or through a gap junction.In short, pericytes induce endothelial maturation and a tighter barrier function, which supports the function against the hypoxic injury. Intercellular communication might be important to keep the BBB functional and stabilize in hypoxia. (C) 2004 Elsevier B.V. All rights reserved.