A Novel Method for Assessing Cerebral Edema, Infarcted Zone and Blood-Brain Barrier Breakdown in a Single Post-stroke Rodent Brain
A Novel Method for Assessing Cerebral Edema, Infarcted Zone and Blood-Brain Barrier Breakdown in a Single Post-stroke Rodent Brain
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DOI：
10.3389/fnins.2019.01105
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发表时间：
2019-10-16
影响因子：
4.3
通讯作者：
Boyko, Matthew
中科院分区：
文献类型：
--
作者：
Kuts, Ruslan;Frank, Dmitry;Boyko, Matthew
中风是全球发病率和死亡率的主要原因。大脑中动脉闭塞（MCAO）历来是模拟缺血性卒中的最常见动物模型。MCAO后神经损伤的程度通常通过脑水肿、梗死区和血脑屏障（BBB）通透性来测量。这些方法的一个重要局限性是，每次测量都必须使用不同的大脑。在这里，我们研究了另一种方法来测量脑水肿，梗死区和血脑屏障通透性以下MCAO在同一组脑样本。将96只大鼠随机分为3个实验组。第1组（n = 27）用于评价MCAO后大鼠（n = 17）与假手术对照组（n = 10）的梗死区和脑水肿。第2组（n = 27）用于评价MCAO后大鼠（n = 15）与假手术对照组（n = 10）的BBB破坏。在第3组（n = 42）中，在MCAO后大鼠（n = 26）与假手术对照组（n = 16）的同一组脑切片中测量所有三个参数。通过测量有和没有应用蓝色滤光片的梗死区来确定伊文思蓝对2，3，5-氯化三苯基四氮唑（TTC）染色测量梗死区准确性的影响。还评估了各种浓度的TTC（0、0.05、0.35、0.5、1和2%）对测量BBB渗透性的准确性的影响。与假手术对照组相比，MCAO后大鼠的梗死体积（p < 0.01）、脑水肿（p < 0.01）和BBB破坏（p < 0.01）增加，无论是单独测量还是在同一组脑样品中一起测量。伊文思蓝对测量梗死体积有影响，通过在扫描的脑切片上应用蓝色滤光片可以使其最小化。与TTC中孵育的脑组织样品相比，不含TTC的脑组织样品的伊文思蓝外渗指数无差异。我们的研究结果表明，测量脑水肿，梗死区和血脑屏障通透性以下MCAO可以准确地测量在同一组脑样本。
Stroke is a major cause of global morbidity and mortality. Middle cerebral artery occlusion (MCAO) has historically been the most common animal model of simulating ischemic stroke. The extent of neurological injury after MCAO is typically measured by cerebral edema, infarct zone, and blood-brain barrier (BBB) permeability. A significant limitation of these methods is that separate sets of brains must be used for each measurement. Here we examine an alternative method of measuring cerebral edema, infarct zone and BBB permeability following MCAO in the same set of brain samples. Ninety-six rats were randomly divided into three experimental groups. Group 1 (n = 27) was used for the evaluation of infarct zone and brain edema in rats post-MCAO (n = 17) vs. sham-operated controls (n = 10). Group 2 (n = 27) was used for the evaluation of BBB breakdown in rats post-MCAO (n = 15) vs. sham-operated controls (n = 10). In Group 3 (n = 42), all three parameters were measured in the same set of brain slices in rats post-MCAO (n = 26) vs. sham-operated controls (n = 16). The effect of Evans blue on the accuracy of measuring infarct zone by 2,3,5-triphenyltetrazolium chloride (TTC) staining was determined by measuring infarct zone with and without an applied blue filter. The effects of various concentrations of TTC (0, 0.05, 0.35, 0.5, 1, and 2%) on the accuracy of measuring BBB permeability was also assessed. There was an increase in infarct volume (p < 0.01), brain edema (p < 0.01) and BBB breakdown (p < 0.01) in rats following MCAO compared to sham-operated controls, whether measured separately or together in the same set of brain samples. Evans blue had an effect on measuring infarct volume that was minimized by the application of a blue filter on scanned brain slices. There was no difference in the Evans blue extravasation index for the brain tissue samples without TTC compared to brain tissue samples incubated in TTC. Our results demonstrate that measuring cerebral edema, infarct zone and BBB permeability following MCAO can accurately be measured in the same set of brain samples.