Quantitation of Early Vascular Leakage after Stroke

描述（由申请方提供）：血脑屏障破坏是许多神经系统疾病的常见因素;大量文献证明了血脑屏障破坏的炎症过程与随后的神经组织死亡之间的时间和/或空间相关性（PAS-04-072）。此外，最近对中风患者的纵向神经成像研究表明，缺血后早期血脑屏障破坏与出血性转化和不良临床结果的可能性增加相关（Latour等，Ann Neurol，2004，出版中）。然而，血脑屏障破坏的时间和空间模式的特点很差，触发这些过程的潜在机制仍然知之甚少。我们建议开发一种新的实验方法来量化成年大鼠实验性中风后血脑屏障的破坏。这些研究的主要目标是开发定量方法来测量血脑屏障的破坏，并保存组织形态学，用于组织缺氧、血栓形成和低血流量的相关和定量免疫细胞化学研究。在这个R21开发项目中，我们将使用可逆性大脑中动脉闭塞（MCAo）的大鼠卒中模型来开发缺血异质性的测量方法，并测试这种异质性的潜在机制。1.发展局部微血管血脑屏障破坏的定量方法。2.通过对组织缺氧、血管反应性和微血栓形成的标记物进行共染色，证实FITC-葡聚糖血脑屏障破坏的区域与缺血性病理学相关。3.证明FITC-葡聚糖血脑屏障破坏与微血管中低血流量的体内测量相关。

DESCRIPTION (provided by applicant): Blood brain barrier breakdown is a common element in many neurological diseases; an extensive literature demonstrates temporal and/or spatial associations between the inflammatory processes that underly blood brain barrier breakdown and subsequent death of neural tissue (PAS-04-072). Further, recent longitudinal neuro-imaging studies in stroke patients indicate that early post-ischemic blood brain barrier breakdown correlates with an increased likelihood for hemorrhagic transformation and poor clinical outcome (Latour et al, Ann Neurol, 2004, in press). However, the temporal and spatial patterns of blood brain barrier break down are poorly characterized, and the underlying mechanisms that trigger these processes remain poorly understood. We propose to develop a new experimental method to quantify blood brain barrier breakdown after experimental stroke in the adult rat. The major goal of these studies is to develop quantitative methods to measure blood brain barrier breakdown and also preserve tissue morphology for correlative and quantitative immunocytochemical studies of tissue hypoxia, thrombosis and low blood flow. In this R21 developmental project, we will use a rat stroke model of reversible middle cerebral artery occlusion (MCAo) to develop measures of ischemic heterogeneity and test potential mechanisms underlying that heterogeneity. 1. Develop methods for the quantitation of local microvascular blood brain barrier break down. 2. Validate that areas of FITC-dextran blood brain barrier breakdown correlate with ischemic pathology by co-staining for markers of tissue hypoxia, vessel reactivity, and mircrothrombosis. 3. Validate that FITC-dextran blood brain barrier breakdown correlates with in vivo measures of low blood flow in microvessels.