脑缺血再灌注损伤导致大鼠神经血管单元破坏  

Cerebral ischemia-reperfusion injury results in the destructionof neurovascular units in rats

作　　者：张海默 高啸天 张孝祖 肖培伦 于淼 蒋景英 王晓莉 ZHANG Haimo;GAO Xiaotian;ZHANG Xiaozu;XIAO Peilun;YU Miao;JIANG Jingying;WANG Xiaoli(School of Medical Imaging,Shandong Second Medical University;Department of Human Anatomy,School of Basic Medical Sciences,Shandong Second Medical University,Weifang 261053;Faculty of Medical Sciences and Engineering,Beijing University of Aeronautics and Astronautics,Beijing 100191,China)

机构地区：[1]山东第二医科大学医学影像学院 [2]山东第二医科大学基础医学院人体解剖学教研室,潍坊261053 [3]北京航空航天大学医学科学与工程学院,北京100191

出　　处：《神经解剖学杂志》2024年第6期724-732,共9页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(82071888);山东省自然科学基金(ZR2024MH067);潍坊市科技发展计划项目(2021YX062);山东省教育厅2019年高等学校青创人才引育计划(鲁教人字[2019]6号)。

摘　　要：目的:基于多模态MRI结合多种组织学技术评估脑缺血再灌注损伤(CIRI)大鼠神经血管单元(NVU)损伤,为临床上CIRI提供可视化依据。方法:随机将28只健康雄性Sprague-Dawley大鼠分为假手术(Sham)组、CIRI组。模型建立24 h后,大鼠进行头部T2加权成像、弥散加权成像、水提取相位对比自旋标记-磁共振成像(MRI)及脑血流血氧监测。HE染色观察大脑皮质神经细胞形态变化。免疫荧光染色及透射电镜观察CIRI后NVU的变化。免疫组化染色与Western Blot检测大鼠大脑皮质claudin-5、ZO-1及基质金属蛋白酶9(MMP-9)表达。结果:MRI结果显示,CIRI组脑组织水肿,血流灌注量显著降低、BBB通透性较Sham组显著增加(P<0.01)。脑血流血氧监测结果显示,损伤侧大脑皮质血红蛋白氧饱和度显著低于Sham组(P<0.05)。HE染色结果显示,损伤侧大脑皮质神经细胞排列紊乱,核固缩。免疫荧光染色结果显示损伤侧大脑皮质NeuN、CD31阳性细胞数显著低于Sham组(P<0.01),大脑皮质及纹状体TUNEL阳性细胞数显著多于Sham组(P<0.05)。透射电镜观察结果显示损伤侧大脑皮质内皮细胞紧密连接破坏,神经元核固缩、星形胶质细胞胞体水肿。免疫组化染色结果显示损伤侧大脑皮质claudin-5、ZO-1阳性细胞数显著低于Sham组,MMP-9表达显著高于Sham组(P<0.01)。结论:大鼠CIRI可导致NVU损伤,破坏BBB,加重脑损伤,多模态MRI结合多种组织病理学技术可有效评估CIRI后NVU损伤。Objective:The neurovascular unit(NVU)damage in rats with cerebral ischemia-reperfusion injury(CIRI)was evaluated based on multimodal MRI combined with multiple histological techniques to provide visual evidence for clinical assessment of CIRI.Methods:28 healthy male Sprague-Dawley rats were randomly divided into sham-operated(Sham)group and CIRI group.At 24 h after modeling,rats underwent head T2-weighted imaging,diffusion-weighted imaging,water extraction with phase-contrast arterial spin labeling magnetic resonance imaging(MRI),and cerebral blood bleeding oxygen monitoring.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of nerve cells in cerebral cortex.Immunofluorescence staining and transmission electron microscopy were used to observe the changes of NVU after CIRI.The endothelial tight junction proteins(claudin-5 and ZO-1)and matrix metalloproteinase-9(MMP-9)were detected by immunohistochemical staining and Western Blot analysis,to evaluate blood-brain-barrier(BBB)disruption in CIRI rats.Results:The MRI results showed that the brain tissue of the CIRI group was edematous,and the blood perfusion significantly decreased and the BBB permeability significantly increased compared with that of the Sham group(P<0.01).The results of cerebral blood hemoglobin oxygen monitoring showed that the hemoglobin oxygen saturation of the cerebral cortex on the injured side was significantly lower than that of the Sham group(P<0.05).HE staining showed the neuronal cells of the injured cerebral cortex were disordered and the nuclei shrank.Immunofluorescence staining results showed the number of NeuN and CD31 positive cells in the cerebral cortex of the injured side was significantly lower than that in the Sham group(P<0.01),and the number of TUNEL positive cells in the cerebral cortex and striatum was significantly higher than that in the Sham group(P<0.05).Transmission electron microscopy revealed the tight junctions of cerebral cortical endothelial cells were damaged,neuronal nucleus contraction,and

关 键 词：多模态磁共振成像 脑缺血再灌注损伤 神经血管单元 血脑屏障 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]