New method for the quantitative assessment of axonal damage in focal cerebral ischemia
New method for the quantitative assessment of axonal damage in focal cerebral ischemia
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DOI：
10.1097/00004647-200209000-00005
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发表时间：
2002-09-01
影响因子：
6.3
通讯作者：
Macrae, IM
中科院分区：
文献类型：
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作者：
Imai, H;McCulloch, J;Macrae, IM
为了全面评估神经保护药物的疗效，需要对灰质和白质的损伤进行量化。尽管脑缺血后神经元核周损伤的量化方法已经建立，但对轴突损伤的评估一直是有限的。本文介绍了一种定量测定大鼠大脑中动脉(MCA)闭塞后轴突损伤的新方法，并将其应用于抗氧化剂Ebselen的研究。该方法基于免疫组织化学检测缺血区变形、肿胀的轴突中淀粉样前体蛋白(APP)的积聚。对大脑中动脉区域内65个富含轴突的部位进行了轴突肿胀中APP的存在(评分为1)或缺失(评分为0)的评估。各个部位的得分是在预定义的神经解剖区域(例如，胼胝体)、立体定位水平或半球APP总分中求和的。评委内和评委间的重复性都很高(r分别为0.87和0.80)。Ebselen(1 mg·kg~(-1)·h~(-1)，静脉注射)可显著减少神经元损伤体积(24%，P<0.002)和轴突损伤(APP总分从27[23.9~35.1，95%CI]降至21.5[18.2~23.3]，P=0.002)。总之，我们描述了一种可靠且普遍适用的方法来评估有髓纤维束的病理特征，该方法对检测药物对轴突损伤的影响是敏感的。
Quantification of damage in both grey and white matter is required for comprehensive assessment of neuroprotective drug efficacy. Although methods for quantification of neuronal perikaryal damage after ischemia are Well established, assessment of axonal damage has been limited. This article describes a new method for quantitation of axonal injury after middle cerebral artery (MCA) occlusion in rats and its application to the study of the antioxidant ebselen. The methodology is based on immunohistochemical detection of amyloid precursor protein (APP) accumulation in deformed, swollen axons in zones of ischemia. Sixty-five axon-rich sites throughout the MCA territory are assessed for the presence (scored 1) or absence (scored 0) of accumulated APP in axonal swellings. Scores for individual sites are summated in predefined neuro-anatomic regions (e.g. corpus callosum), stereotaxic levels, or for a total hemisphere APP score. Both intra-rater and inter-rater reproducibility were high (r = 0.87 and 0.80, respectively). Ebselen (1 mg kg(-1) hr(-1), intravenously) significantly reduced the volume of neuronal perikaryal damage (24%, P < 0.01) and axonal damage (total APP score reduced from 27 [23.9 to 35.1, 95% CI] to 21.5 [18.2 to 23.3], P = 0.002 With ebselen treatment). In conclusion, a robust and generally applicable method is described for assessing pathologic features in myelinated fiber tracts that is sensitive for detection of drug effects on axonal damage.