急性缺氧对小鼠大脑皮质和海马的时间依赖性损伤  

Time-dependent injury of mouse cerebral cortex and hippocampus by acute hypoxia

作　　者：石华香 周梦玮 周琥 张晶鑫 史卫国 王丽韫 SHI Hua-Xiang;ZHOU Meng-Wei;ZHOU Hu;ZHANG Jing-Xin;SHI Wei-Guo;WANG Li-Yun(School of Chemical Engineering and Technology,Tianjin University,Tianjin 300350,China;State Key Laboratory of Toxicology and Medical Countermeasures,Institute of Toxicology and Pharmacology,Academy of Military Medicine Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]天津大学化工学院,天津300350 [2]军事科学院军事医学研究院毒物药物研究所,抗毒药物与毒理学国家重点实验室,北京100850

出　　处：《生理学报》2022年第2期145-154,共10页Acta Physiologica Sinica

基　　金：supported by the National Science and Technology Major Project (No.2018ZX09301-006)。

摘　　要：本文旨在研究急性缺氧对小鼠大脑皮质和海马组织的细胞损伤作用及其机制。利用密闭玻璃罐构建急性缺氧小鼠模型,采用激光散斑成像仪检测小鼠不同缺氧时间窗的脑血流变化,用总超氧化物歧化酶(total superoxide dismutase, T-SOD)和丙二醛(malondialdehyde, MDA)试剂盒检测大脑皮质和海马组织的氧化应激,用组织免疫荧光染色法检测小鼠大脑皮质和海马组织神经炎性反应,用一步法TUNEL细胞凋亡试剂盒检测大脑皮质和海马组织中神经元的凋亡。结果显示,与未缺氧(0min)小鼠相比,缺氧30 min小鼠脑血流显著降低,大脑皮质和海马组织CD68^(+)/Iba1^(+)小胶质细胞百分比显著增加,且神经元凋亡显著增多。与缺氧30 min小鼠相比,缺氧60 min小鼠脑血流相对值显著降低,大脑皮质MDA含量显著增加,大脑皮质和海马中CD68;/Iba1;小胶质细胞百分比显著增加,神经元凋亡显著增多。上述结果提示,急性缺氧诱发小鼠脑组织损伤程度具有时间依赖性,而氧化应激和神经炎性作用是重要的损伤机制。The aim of this study was to investigate the harmful effects of acute hypoxia on mouse cerebral cortex and hippocampus and the underlying mechanism. Mouse model of acute hypoxia was constructed by using a sealed glass jar. Laser speckle contrast imaging was used to detect the changes of cerebral blood flow after different time duration of hypoxia. Total superoxide dismutase(T-SOD) and malondialdehyde(MDA) assay kits were used to detect oxidative stress in cerebral cortex and hippocampus. Immunofluorescent staining was used to detect neuroinflammatory response of microglia in the cerebral cortex and hippocampus. One-step TUNEL method was used to detect neuronal apoptosis. The results showed that, compared with non-hypoxia(0 min hypoxia) group,30 min hypoxia group exhibited decreased cerebral blood flow, higher percentage of CD68^(+)/Iba1^(+)microglia, and increased neural apoptosis in the cerebral cortex and hippocampus. Compared with 30 min group, 60 min hypoxia group showed significantly decreased cerebral blood flow, increased MDA content in the cortex, as well as greater percentage of CD68+/Iba1+ microglia and neuronal apoptosis in the cerebral cortex and hippocampus. These results suggest that acute hypoxia damages brain tissue in a time-dependent manner and the oxidative stress and neuroinflammation are important mechanisms.

关 键 词：脑血流 缺氧 激光散斑成像 氧化应激 免疫荧光 凋亡 

分 类 号：Q42[生物学—神经生物学]