摘要
目的建立一种简便、可靠、重复性好、与新生儿期临床表现及病理生理过程相似的缺氧缺血性脑病（hypoxic-ischemic encephalopathy，HIE）的动物模型。方法7日龄Sprague-Dawley大鼠180只，随机分为空白对照组、实验对照组、缺氧8 min组、缺氧10 min组、缺氧12 min组、缺氧14 min组，每组30只。缺氧8 min组、缺氧10 min组、缺氧12 min组及缺氧14 min组大鼠采用5%利多卡因局部麻醉后，钝性分离气管，分别用血管夹夹闭8、10、12、14 min。实验对照组仅钝性分离气管，术毕创面消毒，缝合。空白对照组不给予任何处理。缺氧达14 min后，存活率明显降低，故缺氧14 min组大鼠未进一步行组织形态学及行为学检测。建模后12 h行脑组织切片苏木精-伊红染色观察。建模后3 d，测量脑重，进行脑组织末端脱氧核苷转移酶介导的dUTP缺口末端标记染色检测神经细胞凋亡情况。2月龄时行Morris水迷宫实验检测大鼠认知功能损害情况。采用单因素方差分析对数据进行方差分析后，方差齐两两比较用SNK检验，方差不齐两两比较用Dunnett＇s T3检验。结果缺氧后大鼠出现全身肤色青紫,意识丧失，进而全身苍白,大小便失禁，四肢、尾巴抽搐等行为学表现。缺氧8 min组、缺氧10 min组及缺氧12 min组均可见到缺血坏死、组织出血、大量神经元细胞胞核皱缩及深染等缺氧后表现。建模后3 d，缺氧8 min组、缺氧10 min组及缺氧12 min组大鼠脑重均低于空白对照组及实验对照组[分别为（1.16±0.07）、（1.04±0.06）、（0.97±0.12）、（1.31±0.06）及（1.28±0.09） g，F=36.437，P〈0.001]，颞叶皮层[分别为（22.83±4.52）、（30.25±3.02）、（39.18±5.04）、（7.96±2.24）及（8.86±2.49）个/400倍视野，F=164.532，P〈0.001]及海马CA3区神经细胞凋亡数量[分别为（14.63±2.26）、（20.25±3.02）、（24.81±1.98）、（4.75±2.66）及（6.67±1.78）个/4
ObjectiveTo establish a simple, reliable and reproducible animal model of neonatal hypoxic-ischemic encephalopathy （HIE） with similar clinical pathological process to neonates.MethodsSeven days after birth, 180 Sprague-Dawley （SD） rats were randomly divided into six groups： blank control group, experimental control group and four hypoxia groups （8, 10, 12 and 14 min hypoxia groups）. Those in the experimental groups were locally anesthetized with 5% lidocaine to separate their tracheas through blunt dissection, followed by tracheal clamping with vascular clamp for 8, 10, 12 and 14 min, respectively. Rats in the experimental control group were only treated with blunt dissection of trachea. No intervention was given to the blank control group. Due to significant reduction in rat survival rate after 14 min of hypoxia, no further morphological or behavioral examination was performed in this group. Rat brain tissue sections were stained with hematoxylin-eosin （HE） 12 h after modeling. Three days after modeling, the rat brain was weighted and the apoptosis of neural cells was detected with terminal deoxynucleotidyl transferase（TdT） mediated dUTP nick end labeling （TUNEL）. Morris water maze was used to screen cognitive impairment in these rats at the age of two months. One-way analysis of variance was used for statistical analysis. SNK test and Dunnett ＇s T3 test were performed to compare homogeneous and non-homogeneous data between groups.ResultsSystemic cyanosis, loss of consciousness, paled body, urinary and fecal incontinence, twitching of the limbs and tail and other abnormal behavior were induced by hypoxia. Ischemic necrosis, bleeding, nucleus shrinkage in a large number of neurons and hyperchromatic nuclei were observed in the 8, 10 and 12 min hypoxia groups. Three days after modeling, brain weights of rats in the 8, 10 and 12 min hypoxia groups were lower than those of the blank control group and experimental control group [（1.16±0.07）, （1.04±0.06）, （0.97±0.12）, （1.31
出处
《中华围产医学杂志》
CAS
CSCD
2017年第12期882-887,共6页
Chinese Journal of Perinatal Medicine
基金
福建省自然科学基金（2014J01276）
福建省卫生计生委青年科研基金（2014-1-18）
关键词
缺氧缺血
脑
疾病模型
动物
气管
收缩
Hypoxia-ischemia, brain
Disease models, animal
Trachea
Constriction
