反复缺血再灌注制备小鼠血管性痴呆模型的评价

The evaluation on the mouse model of vascular dementia by repeated ischemia-reperfusion method

目的对应用反复缺血再灌注法制备的小鼠血管性痴呆(VD)模型进行评价,为VD的基础研究提供较为理想的动物模型。方法50只小鼠随机分为模型组(n=30)和假手术对照组,模型组采用双侧颈总动脉丝线结扎方法、经连续3次反复缺血再灌注,制备VD模型;对照组仅分离双侧颈总动脉,但不阻断血流。测试2组小鼠的学习记忆成绩;光学显微镜下观察海马CA1区神经细胞形态学变化,并采用免疫组化技术观测该区胆碱乙酰转移酶(ChAT)表达的变化。结果跳台试验中模型组小鼠术后第29d的反应时间(125.4±32.5)s较对照组(24.9±2.9)s明显延长(P<0.01),错误次数增多(P<0.01);术后第30d的潜伏时间(80±29)s较对照组(200±37)s明显缩短(P<0.01),错误次数增多(P<0.01)。水迷宫试验中模型组小鼠术后第29d、30d游完全程时间[(143±17)s,(162±11)s]较对照组[(68±8)s,(52±5)s]明显延长(P<0.01),错误次数增多(P<0.01)。海马CA1区ChAT免疫组化观察中模型组阳性神经元面密度值为0.086±0.009,较假手术组(0.123±0.015)明显降低(P<0.01)。结论反复缺血再灌注法制备的小鼠模型是研究血管性痴呆较为理想的动物模型。

Objective To evaluate the mouse model of vascular dementia （VD） established by repeated ischemia-reperfusion method, in order to provide an available animal model for the basic research of VD. Methods 50 Kunming mice were randomly derided into two groups： the control group and VD group. In VD group, mice were subjected for continuously repeated three times ischemia-reperfusion through the ligation of the bilateral common carotid arteries to establish VD model. The behavioral abnormalties were investigated by step-down test and water-maze test. The changes of neurons in hippocampal CAI were observed through light microscope and choline acetyltransferasc （CHAT） in neurons of hippocampus were observed by immunohistochemistry technique. Results In step-down test, the response period of mice in the VD group （125.4 ± 32.5s） was prolonged evidently than control group （24.9± 2.9s） （ P 〈 0.01 ）, and the error times increased （ P 〈 0.01 ） on the 29th day after operation, the latent period in the VD group （80±29s） was deceased significantly than control gloup （200±37s） （ P 〈 0.01） ,and the error times increased （ P 〈 0.01 ） on the 30th day after operation. In water-maze test, the swimming time of mice in VD group was （ 143 ± 17）s and （ 162 ± 11 ）s on the 29th and 30th day after operation, which was evidently prolonged than that of control group （68s ± 8s and 52s ± 5s respectively） （ P 〈 0.01 ） ; and the nttmber of number increased （ P 〈 0.01） .The neurons were damaged and the expression of ChAT decreased（0.086 ± 0.009 vs 0. 123 ± 0.015, P 〈 0.01） .Conclusions The model established by repeated ischemia-reperfusion injury is a reliable VD model, which imitated the clinical intelligent dysfunction of VD.