作者：郭文娟，孙庆玲，李婷，魏明清，倪敬年，时晶

单位：

北京中医药大学东直门医院脑病三科

Units：

Department of Neurology, Dongzhimen Hospital, Beijing University of Chinese Medicine;

关键词：

慢性脑低灌注; 血管性认知障碍; 双侧颈总动脉狭窄; 小鼠;

Keywords：

Chronic cerebral hypoperfusion;Vascular cognitive impairment;Bilateral common carotid artery stenosis;Mice;

CLC：

DOI：

10.12114/j.issn.1008-5971.2022.00.299

Funds：

国家自然科学基金面上项目（8207151194）； 中央高校基本科研业务费（2019-JYB-TD-007）； 岐黄学者基金；

摘要：

目的 通过改良微弹簧双侧颈总动脉狭窄（BCAS）法构建慢性脑低灌注（CCH）小鼠模型，并观察其认知功能。方法 本实验时间为2022年4—5月。选取30只雄性C57BL/6J小鼠，随机分为假手术组和模型组，各15只。模型组小鼠通过改良微弹簧BCAS法构建CCH小鼠模型，主要使用直径0.08 mm的非吸收外科缝合线，将直径0.18 mm的钢丝与颈总动脉（CCA）牢固结扎，然后缓慢抽出钢丝。假手术组小鼠仅分离两侧CCA，随后缝合消毒。术后第1天进行MRI扫描；术后第1天、1个月和2个月，分别从假手术组和模型组随机选取5只小鼠，检测其脑血流量（CBF）及低灌注区占比；两组小鼠均于术后2个月进行Morris水迷宫实验，以检测其实验第1～5天游泳距离、逃避潜伏期及实验第6天60 s内穿过平台象限的次数。结果 T2WI显示，术后第1天两组小鼠均未发生急性脑梗死及其他异常改变。术后第1天、1个月、2个月，模型组小鼠CBF少于假手术组，低灌注区占比高于假手术组（P<0.05）；术后1个月、2个月，模型组小鼠CBF多于术后第1天，低灌注区占比低于术后第1天（P<0.05）。实验第3、4、5天，模型组小鼠游泳距离长于假手术组（P<0.05）。实验第1、2、3、4、5天，模型组小鼠逃避潜伏期长于假手术组（P<0.05）。实验第6天，模型组小鼠60 s内穿过平台象限的次数多于假手术组（P<0.05）。结论 通过改良微弹簧BCAS法构建的CCH小鼠模型弥补了微弹簧无法进行活体MRI扫描的缺陷，且能维持低灌注状态，小鼠存在学习和记忆能力受损，这为探究CCH发病机制提供了新的建模思路。

Abstract：

Objective To construct the mouse model of chronic cerebral hypoperfusion（CCH） by modified microcoil bilateral common carotid artery stenosis（BCAS） method, and to observe its cognitive function. Methods From April to May 2022, 30 male C57BL/6J mice were selected and randomly divided into the Sham group and the model group, with 15 in each group. The mouse in the model group was constructed the CCH mouse model by the modified microcoil BCAS method. Common carotid artery（CCA） and a steel wire with a diameter of 0.18 mm were firmly ligated with a non-absorbable surgical suture with a diameter of 0.08 mm, and then the steel wire was slowly removed. The mice in the Sham group only separated the CCA on both sides, and then sutured and disinfected. MRI scan was performed on the 1st day after operation; 5 mice were randomly selected from the Sham group and the model group on the 1st day and at 1 month and 2 months after operation, respectively, and their cerebral blood flow（CBF） and percentage of the hypoperfused area were detected. The two groups of mice were underwent Morris water maze test at 2 months after operation to detect the swimming distance and escape latency from the 1st to the 5th day of the experiment and the times of crossing the platform quadrant within 60 s on the 6th day of the experiment. Results T2WI scan showed that acute cerebral infarction and other abnormal changes did not occur in both groups of mice on the 1st day after operation. On the 1st day and at 1 month and 2 months after operation, the CBF of mice in the model group was less than that in the Sham group, and the percentage of the hypoperfused area was higher than that in the Sham group（P < 0.05）; at 1 month and 2 months after operation, the CBF of mice in the model group was more than that on the 1st day after operation, and the percentage of the hypoperfused area was lower than that on the 1st day after operation（P < 0.05）. On the 3rd, 4th and 5th day of the experiment, swimming distance in the model group was longer than that in the Sham group（P < 0.05）. On the 1st, 2nd, 3th, 4th and 5th day of the experiment, escape latency in the model group was longer than that in the Sham group（P < 0.05）. On the 6th day of the experiment, the times of crossing the platform quadrant within 60 s were more than those in the Sham group（P < 0.05）. Conclusion The CCH mouse model constructed by the modified microcoil BCAS method makes up for the defect that the micro-spring cannot perform in vivo MRI scanning, and can maintain a low perfusion state. The mice have impaired learning and memory ability, which provides a new modeling idea for exploring the pathogenesis of CCH.

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