IN VITRO BLOOD BRAIN BARRIER

我们的目标是开发一种血液的体外模型 血管生成的主要假说通过的脑屏障 可以进行脑水肿的检测。脑水肿伴随着许多 不同的病理侮辱中枢神经系统，是一个质数 最终发病率或死亡率的决定因素 与许多这样的侮辱有关。有几个假设 被提出用来解释血脑的紊乱 血管源性脑形成之前的屏障 浮肿。花生四烯酸代谢是大多数 这些假说，但准确地描述了 所涉及的代谢物、它们的起源和/或作用部位，以及 他们的行动机制在很大程度上仍未确定。 独一无二的，小鼠脑内皮细胞和血管平滑肌 和大鼠C6胶质瘤细胞在实验室中可用于 这些假说的体外探索。初步研究 提示脑内皮细胞的某些特性是 在培养中保持(即，对某些 大分子)，而其他的则需要大脑的共培养 血管内皮细胞与C6胶质瘤细胞的表达(即， γ-GTP活性和极性对氨基酸运输的影响)。 进一步研究这一特征的体外血脑 屏障模型将包括：1)渗透率研究 与大相对分子质量和低辛醇/水分配比 共效率溶质，2)超微结构探针用于分析 囊泡转运，3)跨内皮细胞的测量 电阻，以及4)更广泛的共培养研究 评估血脑所需的细胞间相互作用 屏障的区分与维护。 花生四烯酸代谢的细胞类型特异性研究 主要利用薄层层析，高性能 高效液相色谱和放射免疫分析将表征 二十烷类化合物的特定代谢部位。初步研究 揭示血管内皮细胞和血管内皮细胞的区别。 关于脑血管二十烷类新陈代谢的知识将被 用来评估其在体外模型中的作用 血脑屏障发育较早。

Our objective is to develop an in vitro model of the blood brain barrier through which major hypotheses of vasogenic cerebral edema may be tested. Cerebral edema accompanies many different pathological insults to the CNS and is a prime determining factor of the ultimate morbidity or mortality associated with many of these insults. Several hypotheses have been proposed to explain the disruption of the blood brain barrier which precedes the formation of vasogenic cerebral edema. Arachidonic acid metabolism is a central issue in most of these hypotheses, but precise characterization of the metabolites involved, their sites of origin and/or action, and the mechanisms of their actions remains largely undetermined. Uniquely, mouse cerebral endothelium and vascular smooth muscle and rat C6 glioma cells are available in the laboratory for in vitro exploration of these hypotheses. Preliminary studies suggest that some cerebral endothelial properties are maintained in culture (i.e, impermeability to certain macromolecules), while others require co-culture of cerebral endothelium with C6 glioma cells for their expression (i.e., Gamma-GTP activity and polarity to amino acid transport). Further studies to characterize this in vitro blood brain barrier model will include: 1) permeability studies with small versus large M.W. and low versus high octanol/water partition co-efficient solutes, 2) ultrastructural probes for analysis of vesicular transport, 3) measurement of transendothelial electrical resistance, and 4) more extensive co-culture studies to assess intercellular interactions required for blood brain barrier differentiation and maintenance. Cell type specific studies of arachidonic acid metabolism utilizing primarily thin layer chromatography, high performance liquid chromatography, and radioimmunoassay will characterize sites of specific eicosanoid metabolism. Preliminary studies reveal differences between endothelium and smooth muscle. Knowledge of cerebrovascular eicosanoid metabolism will then be utilized to assess its effects on the in vitro model of the blood brain barrier developed earlier.