Department of Medical Pharmacology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan.
Department of Functional Pathology, Shimane University Faculty of Medicine, Izumo, Japan.
Cell Mol Neurobiol. 2022 Jan;42(1):243-253. doi: 10.1007/s10571-020-00917-z. Epub 2020 Jul 9.
The blood-brain barrier (BBB) comprises three cell types: brain capillary endothelial cells (BECs), astrocytes, and pericytes. Abnormal interaction among these cells may induce BBB dysfunction and lead to cerebrovascular diseases. The stroke-prone spontaneously hypertensive rat (SHRSP) harbors a defective BBB, so we designed the present study to examine the role of these three cell types in a functional disorder of the BBB in SHRSP in order to elucidate the role of these cells in the BBB more generally. To this end, we employed a unique in vitro model of BBB, in which various combinations of the cells could be tested. The three types of cells were prepared from both SHRSPs and Wistar Kyoto rats (WKYs). They were then co-cultured in various combinations to construct in vitro BBB models. The barrier function of the models was estimated by measuring transendothelial electrical resistance and the permeability of the endothelial monolayer to sodium fluorescein. The in vitro models revealed that (1) BECs from SHRSPs had an inherent lower barrier function, (2) astrocytes of SHRSPs had an impaired ability to induce barrier function in BECs, although (3) both pericytes and astrocytes of SHRSPs and WKYs could potentiate the barrier function of BECs under co-culture conditions. Furthermore, we found that claudin-5 expression was consistently lower in models that used BECs and/or SHRSP astrocytes. These results suggested that defective interaction among BBB cells-especially BECs and astrocytes-was responsible for a functional disorder of the BBB in SHRSPs.
血脑屏障（BBB）由三种细胞类型组成：脑毛细血管内皮细胞（BECs）、星形胶质细胞和周细胞。这些细胞之间的异常相互作用可能导致 BBB 功能障碍，并导致脑血管疾病。易发生中风的自发性高血压大鼠（SHRSP）具有缺陷的 BBB，因此我们设计了本研究，以检查这三种细胞类型在 SHRSP 中 BBB 功能障碍中的作用，以便更普遍地阐明这些细胞在 BBB 中的作用。为此，我们采用了一种独特的 BBB 体外模型，其中可以测试各种细胞组合。这三种类型的细胞均来自 SHRSP 和 Wistar Kyoto 大鼠（WKYs）。然后将它们以各种组合共培养以构建体外 BBB 模型。通过测量跨内皮电阻和内皮单层对荧光素钠的通透性来评估模型的屏障功能。体外模型表明：（1）SHRSPs 的 BECs 具有固有的较低屏障功能，（2）SHRSPs 的星形胶质细胞诱导 BECs 形成屏障功能的能力受损，尽管（3）SHRSPs 和 WKYs 的周细胞和星形胶质细胞在共培养条件下均能增强 BECs 的屏障功能。此外，我们发现使用 BECs 和/或 SHRSP 星形胶质细胞的模型中 Claudin-5 的表达始终较低。这些结果表明，BBB 细胞之间的缺陷相互作用 - 特别是 BECs 和星形胶质细胞 - 是 SHRSPs 中 BBB 功能障碍的原因。
