Protein expression of brain endothelial cell E-cadherin after hypoxia/aglycemia: influence of astrocyte contact
Protein expression of brain endothelial cell E-cadherin after hypoxia/aglycemia: influence of astrocyte contact
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DOI：
10.1016/s0006-8993(99)01778-3
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发表时间：
1999-09-25
期刊：
影响因子：
2.9
通讯作者：
Davis, TP
中科院分区：
文献类型：
--
作者：
Abbruscato, TJ;Davis, TP
血脑屏障（BBB）在保护中枢神经系统（CNS）免受病理条件带来的内稳态变化方面发挥着至关重要的作用。脑血管通透性是脑卒中后脑水肿发生发展的重要因素[M. Plateel、E.泰西耶河Cecchelli，缺氧，大大增加了非特异性运输的血液蛋白质到大脑。神经化学杂志。68（1997）874-877]，并且其功能的任何变化都可能具有有害的神经学后果。最近，研究已经表明，BBB的体外模型对缺氧/血糖缺乏的短时间暴露敏感，并且内皮细胞钙通量的变化可能是缺血应激期间BBB中结构和功能变化的原因[T. J. Abbruscato，T. P. Davis，缺氧/血糖缺乏的组合损害体外BBB。J. Pharmacol. Exp. Ther. 289（1999）668-675]。目前的实验研究了牛脑微血管内皮细胞（BBMEC）中钙依赖性细胞间粘附分子E-cadherin的表达，E-cadherin已被证明对血脑屏障功能很重要[D. K.L. Audus，T. J. Siahaan，十肽对牛脑微血管内皮细胞中细胞粘附的调节。Brain Research 747（1997）103-113]。由于相信星形胶质细胞-内皮细胞相互作用对于维持体内BBB特征是至关重要的，我们尝试优化我们的分离和培养技术以产生适合于研究病理条件的可靠的BBB体外模型。免疫荧光实验显示阳性染色的E-钙粘蛋白，但未能显示任何变化的细胞分布的E-钙粘蛋白后缺氧/无糖皮质激素暴露。此外，用C6条件培养基（CM）培养BBMEC对E-cadherin的定位没有影响。蛋白质印迹实验表明，BBMEC表达E-钙粘蛋白和这种蛋白质是减少在各种缺氧/glycoprotein暴露后，当内皮细胞单独培养或与C6星形胶质瘤生长在一个单独的培养表面上的时间依赖性的方式。当C6星形胶质细胞与内皮细胞直接相对生长时，在两者之间具有多孔膜，我们观察到在缺氧/无糖暴露后E-钙粘蛋白的降低的BBMEC表达的轻微衰减。这项工作表明，哺乳动物脑内皮细胞/星形胶质细胞共培养系统是一个有用的模型，用于研究的病理条件下，血脑屏障的特点是保持。(C)1999 Elsevier Science B. V.保留所有权利。
The blood-brain barrier (BBB) plays a crucial role in protecting the central nervous system (CNS) from any changes in homeostasis brought about by pathological conditions. Cerebrovascular permeability is an important factor in the development of cerebral edema following stroke [M. Plateel, E. Teissier, R. Cecchelli, Hypoxia, dramatically increases the nonspecific transport of blood-bome proteins to the brain. J. Neurochem. 68 (1997) 874-877] and any changes in its function can have detrimental neurological consequences. Recently, research has shown that an in vitro model of the BBB is sensitive to short exposures of hypoxia/aglycemia and that changes in endothelial cell calcium flux may be responsible for structural and functional variations in the BBB during ischemic stress [T.J. Abbruscato, T.P. Davis, Combination of hypoxia/aglycemia compromises in vitro BBB. J. Pharmacol. Exp. Ther. 289 (1999) 668-675]. Present experiments investigated bovine brain microvessel endothelial cell (BBMEC) expression of a Ca2+-dependent cell-cell adhesion molecule, E-cadherin, which has been shown to be important for blood-brain barrier function [D. Pal, K.L. Audus, T.J. Siahaan, Modulation of cellular adhesion in bovine brain microvessel endothelial cells by a decapeptide. Brain Research 747 (1997) 103-113]. Since it is believed that astrocyte-endothelial cell interaction is crucial for maintenance of in vivo BBB characteristics, we have attempted to optimize our isolation and culturing techniques to produce a reliable, in vitro model of the BBB that is suitable to study pathological conditions. Immunofluoresence experiments showed positive staining for E-cadherin, yet failed to show any change in cellular distribution of E-cadherin upon hypoxic/aglycemic exposure. In addition, culturing BBMECs with C6 conditioned medium (CM) had no effect on the localization of E-cadherin. Western blotting experiments showed that BBMECs express E-cadherin and this protein is decreased in a time dependent manner after various hypoxic/aglycemic exposures when endothelial cells are cultured alone or with C6 astrogliomas grown on a separate culture surface. When C6 astrocytes are grown directly opposed to endothelial cells, with a porous membrane between, we observed a slight attenuation in the decreased BBMEC expression of E-Cadherin after hypoxia/aglycemia exposure. This work has shown that the mammalian brain endothelial/astrocyte co-culture system is a useful model for studies of pathological conditions where BBB characteristics are maintained. (C) 1999 Elsevier Science B.V. All rights reserved.