Neonatal Neuroscience, School of Clinical Sciences, University of Bristol, St Michael's Hospital, Southwell Street, Bristol, Avon BS2 8EG, United Kingdom.
Neonatal Neuroscience, School of Clinical Sciences, University of Bristol, St Michael's Hospital, Southwell Street, Bristol, Avon BS2 8EG, United Kingdom; Department of Physiology, Institute of Basic Medical Sciences, University of Oslo, Postboks 1103, Blindern, 0317 Oslo, Norway.
J Neurosci Methods. 2014 Jun 15;230:30-6. doi: 10.1016/j.jneumeth.2014.04.005. Epub 2014 Apr 18.
Neuropathological examination is the classic outcome measure in experimental studies of newborn brain injury to evaluate novel therapies. We have used a graded neuropathology score in an established global model of perinatal hypoxic-ischaemic (HI) injury. We wished to validate the score using cell counting in our model.
32 newborn pigs underwent a 45 min global HI insult then maintained at normothermia (NT, rectal temperature, Trectal 38.5 °C) for 72 h or mild total body hypothermia (HT, Trectal 37.0 °C) combined with selective head cooling for 48 h and subsequently maintained at NT for 24h before brain perfusion fixation. A perinatal pathologist scored haematoxylin and eosin stained 6 μm histological sections for injury in the hippocampus and basal ganglia on a 9-step scale (0.0=no injury, 4.0=>75% injury). We counted the number of healthy neurons in the hippocampus CA1 region and putamen using morphological criteria in eight random, non-overlapping fields from representative sections.
Healthy neuronal cell density correlated with neuropathology score in the hippocampus CA1 (r = -0.74) and in the putamen (r = -0.75) and both measures detected a difference between groups. The correlation coefficients were better for the NT compared to the HT group in both the hippocampus (r = -0.87 vs. -0.53) and putamen (r = -0.77 vs. -0.54).
We have validated a histological neuropathological scoring system in our model of perinatal HI by showing correlation between neuronal cell count and estimated injury.
Our neuropathology score is a valid method to assess brain injury with good reproducibility and sensitivity.
神经病理学检查是评估新生脑损伤实验研究中新型治疗方法的经典结局指标。我们使用一种已建立的围产期缺氧缺血（HI）损伤全脑模型中的分级神经病理学评分。我们希望通过细胞计数来验证我们模型中的评分。
32 只新生仔猪经历 45 分钟的全脑 HI 损伤，然后在正常体温（NT，直肠温度，Trectal 38.5°C）下维持 72 小时，或轻度全身低温（HT，Trectal 37.0°C），同时进行选择性头部冷却 48 小时，然后在 NT 下维持 24 小时，再进行脑灌注固定。一名围产期病理学家使用 9 级评分（0.0=无损伤，4.0=>75%损伤）对苏木精和伊红染色的 6μm 组织切片进行损伤评分，用于海马和基底节。我们使用形态学标准在代表性切片的八个随机、不重叠的区域中计算海马 CA1 区和壳核中的健康神经元数量。
海马 CA1 区（r = -0.74）和壳核（r = -0.75）的健康神经元细胞密度与神经病理学评分相关，两种测量方法均能检测到组间差异。在海马（r = -0.87 比 -0.53）和壳核（r = -0.77 比 -0.54）中，NT 组的相关系数优于 HT 组。
我们通过显示神经元细胞计数与估计损伤之间的相关性，验证了我们的 HI 新生仔猪模型中神经病理学评分系统的有效性。
我们的神经病理学评分是一种评估脑损伤的有效方法，具有良好的重现性和敏感性。
