摘要
目的建立一种新的鼠脑局部蛛网膜下腔注射氧合血红蛋白(OxyHb)的动物模型,探讨OxyHb在体诱导鼠脑细胞凋亡空间分布及时间依赖性规律的研究。方法采集ICR鼠新鲜血液制备OxyHb,建立小鼠脑局部蛛网膜下腔注射OxyHb动物模型。动物被分为实验组(40只)及对照组(35只),实验组注射OxyHb(50μL),对照组注射生理盐水(50μL)于鼠脑蛛网膜下腔,再按处死时间将小鼠分为3h、6h、12h、24h和48h亚组。采用HE染色、透射电子显微镜及原位细胞凋亡检测法(TUNEL)三种技术辨别细胞坏死或凋亡。结果实验组3h OxyHb集中分布于注射点同侧脑表面,6～12h OxyHb积聚于脑基底池及交叉池．24～48h OxyHb已被吸收；实验组鼠脑细胞凋亡主要分布于OxyHb注射区同侧脑皮质及双侧海马区；经HE染色、透射电子显微镜观察显示鼠脑细胞发生凋亡形态学改变；实验组TUNEL阳性细胞较对照组明显增多,且随时间延长凋亡细胞逐渐减少。结论本实验建立的鼠脑局部蛛网膜下腔注射OxyHb的动物模型,方法较简单,重复性好。OxyHb在体可诱导鼠脑细胞凋亡而非坏死,凋亡细胞主要分布于OxyHb注射区同侧脑皮质及双侧海马区,且随时间延长而凋亡细胞逐渐减少,具有空间分布和时间依赖性的规律。
Objective To develop an animal model of oxyhemoglobin （OxyHb） injection into local subarachnoid space in mice, and to investigate the temporal and spatial change of nerve cell apoptosis induced by OxyHb in vivo. Methods The flesh blood from ICR mice was taken to prepare OxyHb. OxyHb （50 μL） was injected into local subarachnoid spsce in mice as experimental group （n=40）, meanwhile, normal saline （50 μL） was injected instead in mice as control group （n=35）. The mice of the 2 groups were subdivided according to different sacrifice time （3, 6, 12, 24 and 48 h after injection）. The cell apoptosis or necrosis was distinguished by microscopy （HE staining）, transmission electron microscopy and TUNEL. Results OxyHb was visible over the brain surface around the injection point in 3 h, in chiasmatic cistern and basilar cistern in 6-12 h, and was absorbed completely within 24-48 h. The cell apoptosis was mainly located in the ipsilateral neocortex around OxyHb-injected region and bilateral hippocampus. The morphological changes of apoptotic brain cells were observed in the experimental group by HE staining and transmission electron microscopy. The number of TUNEL-positive cells showed a significant increase in the experimental group, and the number ofOxyHb-induced apoptotic cells decreased over time. Conclusion This experiment developed a new method of creating an animal model of OxyHb injection into local subarachnoid space in mice. The procedure is simple and reliable. OxyHb may induce apoptosis, but not necrosis of mouse brain cells in vivo. The apoptotic brain cells distributed mainly in the neocortex ipsilateral to the injection and the bilateral hippocampus in mice, and the number of the apoptotic cells decreased in the time and spatial dependent manner.
出处
《中华神经医学杂志》
CAS
CSCD
2006年第9期887-890,共4页
Chinese Journal of Neuromedicine
基金
国家自然科学基金(30270481)
