新生小鼠脑缺氧缺血性脑病模型的制作

A model of hypoxic-ischemic encephalopathy in neonatal mice

目的建立新生C57BL/6小鼠缺氧缺血性脑病(HIE)模型,并讨论模型制作中的相关问题。方法采用健康7日龄C57BL/6新生小鼠,结扎后剪断右侧颈总动脉,置于37℃、8%低氧浓度环境下,分别于30min、60min、90min、120min取出,并于不同时间处死小鼠,观察脑组织病理改变,并进行行为学检测。结果(1)HIE模型组小鼠寻找目标所需时间和路径远长于正常组小鼠,且其与正常组间评分有统计学意义(P<0.05)。(2)随着缺氧时间的延长,HIE小鼠12h内死亡率逐渐增高,当缺氧达90min时,死亡率为10%,缺氧120min时,死亡率高达83%。(3)HIE模型组甲苯胺蓝染色见神经细胞核固缩,着色加深,胞浆Nissl小体明显减少;HE染色可见右侧大脑皮层、海马等区域神经细胞明显减少,尤以海马区为明显,神经细胞变性坏死、胶质细胞反应性增生,形成胶质细胞结节。(4)病理改变与缺氧时间、缺血缺氧(HI)损伤后时间点有关。结论本方法制作新生C57BL/6小鼠HIE模型获得成功。该模型具有进一步推广意义。

Objective To establish a model of hypoxic-ischemic encephalopathy （HIE） in the neonatal C57BL/6 mice. Methods The normal 7-day-old C57BL/6 mice were studied. After unilateral common carotid artery was ligated and cut, the mouse was put in the environment of 37℃ and 8% hypoxia. The brain was collected at different times after injury. The behavioral and neuropathology changes were observed. Results （1）The HIE mice took more time and longer distance to reach the target than the normal mice （P 〈 0. 05 ）. （2）The mortality of the HIE mouse increased with the hypoxia time. The mortality was 10% in 90min and 83% in 120min. （3）Toluidine staining showed karyopyknosis and Nissl body decreased in neural cells in HIE mice. HE staining showed that nerve cells reduction was in evidence in the right cerebral cortex and hippocampus of the HIE mice, especially in the right hippocampus. The characteristic pathological phenomena of HIE, such as nerve cells denaturalization and necrosis and neuralgia cells hyperplasia could be observed. （4）Pathological findings related to the hypoxia and injury time. Conclusion With the property of the cheep price and the easy duplication, the HIE model is successful and worthy of wide use.