新生儿缺氧缺血脑损伤大鼠模型的制备  

Preparation of a rat model of neonatal hypoxic-ischemic encephalopathy

作　　者：庞炜[1] 曹帅帅 李树祎 田雨光[1] 顾为望[1] 

机构地区：[1]南方医科大学实验动物中心,广州510515 [2]广州医科大学口腔教研室,广州510182

出　　处：《中国比较医学杂志》2016年第6期61-66,74,共7页Chinese Journal of Comparative Medicine

基　　金：高新区发展引导专项(2012B011000004);高新区发展引导专项(2010A011200003);广东省科技基础条件建设项目(2012B060300003);广东省科技基础条件建设项目(2013B030300040);广东省科技基础条件建设项目(2013B060300001);广东省科技基础条件建设项目(2010B060500001);国家国际科技合作专项项目(2011DFA33290)

摘　　要：目的在高原低压环境模拟仓内,模拟新生儿在高原环境下缺氧缺血脑损伤的过程,制备不同海拔高度下高原脑损伤大鼠模型。方法 10日龄的SD新生大鼠32只,随机分为4组,即A组(对照组)和3个实验组:B组(2 000 m组)、C组(4 000 m组)、D组(6 000 m组)。对照组大鼠在屏障环境中饲养,实验组大鼠置于高原低压环境模拟仓结合运动的方法制作新生儿高原脑缺氧缺血模型,运动方式为在舱内游泳槽进行60 min/d的游泳运动,且在高原低压环境模拟仓内生活时间每天不得少于20 h。每组大鼠分别在第3、7、11、15天用Zea Longa 5分制评分标准进行行为学评分并且于第15天采集静脉血,在扫描电镜下观察红细胞形态。每组处死后取脑组织进行HE染色和TTC染色。结果 (1)神经病学评分:实验组B组、C组、D组行为学评分与对照组大鼠相比,行为学评分差异显著(P<0.05),D组与对照组相比差异非常显著(P<0.01)。(2)HE染色结果显示:与对照组大鼠相比,实验组大鼠均有炎症细胞浸润,且炎症细胞浸润程度与海拔高度成正相关。(3)TTC染色表明:在高原环境下大鼠的大脑皮质缺血明显。(4)电镜下观察红细胞形态显示:实验组B组呈帽状结构;C组呈不规则形;D组呈锯齿状。结论本研究采用高原低压环境模拟仓结合运动模拟高原环境制作新生儿缺氧缺血性脑病(HIE)的模型,该模型稳定、可靠,比其他方法更符合缺血缺氧脑损伤的发病机理,与临床贴近,可用于相关研究。Objective To simulate the process of hypoxic-ischemic brain injury at high altitude in a simulated cabin with plateau low pressure environment,and to prepare a rat model of cerebral injuries at different high altitudes.Method Thirty-two 0-day-old neonatal SD rats were divided into four groups,namely group A（ control group） and three test groups： group B（ 2000 m group）,group C（ 4000 m group）,and group D（ 6000 m group）. The rats of control group were reared in a barrier environment. The rats of test groups were placed in a simulated cabin with plateau low pressure environment,and to prepare neonatal cerebral ischemia-hypoxia model by sport activities. The sport movements were carried out in the cabin in a swimming groove 60 min / d,and not less than 20 hours a day at high altitude low pressureenvironment. Zea Longa 5 point scale standard was used to determine the behavioral scores at the 3 th 7 th 11 th 15 th days,and samples were collected on the 15 thday to observe red blood cell morphology using HE and 2,3,5-triphenyltetrazolium chloride（ TTC） staining and ultrastructure by scanning electron microscopy. Result（ 1） The neurological scores of the test groups A,B,C were significantly different from that of the control group（ P 0. 05）,and the scores of test group D and control group were very significantly different（ P 0. 01）.（ 2） The histopathological examination using HE staining showed inflammatory cell infiltration in all rats of the test groups,and the extent of inflammatory cell infiltration was positively correlated with the increase of altitude.（ 3） The histopathology with TTC staining revealed prominent ischemia in the cerebral cortex of rats in the plateau hypoxic environment.（ 4） Scanning electron microscopy showed that the rat erythrocytes were cap-like in the group B,irregular in the group C,and zigzag shape in the group D. Conclusions In this study,a rat model of neonatal hypoxic-ischemic encephalopathy（ HIE） is successfully established by hypoxic cabin

关 键 词：新生儿脑缺氧缺血性脑病 TTC染色 扫描电镜 大鼠 

分 类 号：R33[医药卫生—人体生理学]