中国组织工程研究

中国组织工程研究 ›› 2020, Vol. 24 ›› Issue (35): 5625-5631.doi: 10.3969/j.issn.2095-4344.2922

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

成年小鼠缺血性脑卒中后病理及行为功能的变化

梁彦峰1，郝  鹏1，段红梅1，赵  文1，高钰丹1，李晓光1，2，杨朝阳1   

  1. 1首都医科大学神经生物学系，北京市  100069；2北京航空航天大学生物与医学工程学院，生物材料和神经再生北京市实验室，大数据精准医疗高精尖创新中心，北京市  100191

  • 收稿日期:2020-02-25 修回日期:2020-03-03 接受日期:2020-04-23 出版日期:2020-12-18 发布日期:2020-10-17
  • 通讯作者: 杨朝阳，博士，教授，首都医科大学神经生物学系，北京市 100069
  • 作者简介:梁彦峰，男，1992年生，湖北省安陆市人，汉族，2020年首都医科大学毕业，硕士，主要从成年小鼠脑卒中的病理变化及对行为功能的影响研究。
  • 基金资助:
    国家重点研发计划(2017YFC1104002)；国家重点研发计划(2017YFC1104001)；国家自然科学基金(81941011，31730030，31650001，31320103903)；国家自然科学基金(31670988，31971279)；国家自然科学基金(31771053)；北京市科技计划(Z181100001818007)；北京市教育委员会2018年度科技计划重点项目(KZ201810025030)

Pathological and behavioral changes after ischemic stroke in adult mice

Liang Yanfeng1, Hao Peng1, Duan Hongmei1, Zhao Wen1, Gao Yudan1, Li Xiaoguang1, 2, Yang Chaoyang1   

  1. 1Department of Neurobiology, Capital Medical University, Beijing 100069, China; 2School of Biological and Medical Engineering, Beijing Laboratory of Biomaterials and Nerve Regeneration, Big Data Precision Medical Center for Advanced Innovation, Beihang University, Beijing 100191, China

  • Received:2020-02-25 Revised:2020-03-03 Accepted:2020-04-23 Online:2020-12-18 Published:2020-10-17
  • Contact: Yang Chaoyang, MD, Professor, Department of Neurobiology, Capital Medical University, Beijing 100069, China
  • About author:Liang Yanfeng, Master, Department of Neurobiology, Capital Medical University, Beijing 100069, China
  • Supported by:
    the National Key Research and Development Projects of China, No. 2017YFC1104002 and 2017YFC1104001; the National Natural Science Foundation of China, No. 81941011, 31730030, 31650001, 31320103903, 31670988, 31971279 , 31771053; Beijing Science and Technology Plan, No. Z181100001818007; 2018 Key Science and Technology Plan of Beijing Municipal Education Commission, No. KZ201810025030

PDF

369

摘要：

文题释义：

大脑中动脉末端闭塞模型：该模型最先由TAMURA等在大鼠上开发出来，即开颅手术找到大脑中动脉皮质分支并对其进行结扎或电凝，可以引起稳定的局灶性皮质梗死，在很好模拟人类卒中病理状态的同时死亡率较低。但这种手术方式对手术人员的技术以及设备有较高要求，并且需要后期评估才能确定造模成功与否。

缺血性脑卒中：脑卒中分缺血性和出血性脑卒中，缺血性卒中是由于脑的供血动脉狭窄或闭塞，造成脑组织缺血、缺氧性坏死，是中国致死和致残的主要原因。

背景：大鼠大脑中动脉末端闭塞模型可以引起稳定的局灶性皮质梗死，在很好模拟人类卒中病理状态的同时死亡率较低，但对手术人员的技术以及设备有较高要求，并且需要后期评估才能确定造模成功与否。

目的：建立有效稳定简便的缺血性小鼠脑卒中模型；揭示小鼠脑卒中后梗死区及周围区域的病理变化；探索小鼠脑卒中后的行为学改变。

方法：对小鼠末端大脑中动脉进行永久电凝结扎，24 h后用TTC染色明确该模型梗死范围并统计该模型的成功概率；对小鼠大脑中动脉末端闭塞模型后不同时间点(1，3，7，10，14 d)的脑组织切片进行苏木精-伊红染色，观察缺血坏死区的体积在不同时间点的改变；进行胶质纤维酸性蛋白、Iba-1免疫组织化学染色，检测模型小鼠脑损伤后胶质反应及炎症反应变化；最后应用网格足部错误试验和圆柱体试验评价小鼠卒中后感觉运动功能的缺失情况。

结果与结论：①大脑中动脉末端闭塞模型导致局灶性皮质梗死且该新模型死亡率仅9%，成功率达87%；②梗死区域主要位于M1/S1/S2区，梗死范围在闭塞10 d时趋于稳定；③脑卒中后3 d时炎症反应达到高峰，14 d时损伤区周围形成稳定星形胶质瘢痕；④局灶性皮质梗死后小鼠对侧肢体出现明显的感觉及运动的缺失；⑤结果表明，卒中后实验小鼠立即出现感觉及运动功能缺失，且持续至卒中后12周；实验建立的大脑中动脉末端闭塞模型稳定、可靠，梗死范围明确，适合缺血性脑卒中的研究。

ORCID: 0000-0002-5489-3807(梁彦峰)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 脑卒中, 缺血性脑卒中, 大脑中动脉, 末端闭塞, 病理, 行为学, 模型, 鼠

Abstract:

BACKGROUND: A rat model of middle cerebral artery embolization can cause stable focal cortical infarction, and it has a low mortality rate while simulating the pathological state of human stroke. However, there are high requirements on the skills and equipment and a later evaluation is necessary to determine the success of modeling.

OBJECTIVE: To establish an effective, stable and simple ischemic stroke model, to reveal pathological changes in infarcted and surrounding areas after stroke in mice, and to explore behavioral changes after stroke in mice.

METHODS: Permanent electrocoagulation of the distal middle cerebral artery in mice was condcted. Twenty-four hours later, 2,3,5-triphenyltetrazolium chloride staining was used to determine the infarct size of the model and to calculate the success rate of the model. Hematoxylin-eosin staining was performed on brain tissue sections at different time points (1, 3, 7, 10, 14 days) after occlusion of the distal middle cerebral artery in mice to observe volume changes of the ischemic necrosis area with time. Immunohistochemical staining of glial fibrillary acidic protein and Iba-1 was performed to detect the changes of glial reaction and inflammatory reaction after brain injury in model mice. Finally, grid foot error test and cylinder test were used to evaluate the loss of sensorimotor function after stroke.

RESULTS AND CONCLUSION: Focal cortical infarction was caused after distal middle cerebral artery occlusion. The mortality rate of this new model was only 9% and the success rate was 87%. Infarction was mainly located in M1/S1/S2 area, and the infarcted range was stable after 10 days of occlusion. The inflammatory response peaked on the 3rd day of stroke, and a stable astroglial scar was formed around the injury area on the 14th day. After focal cortical infarction, the contralateral limbs of mice showed obvious loss of sensory and motor functions. To conclude, sensory and motor functions deficits occur immediately after stroke in experimental mice and persist until 12 weeks after stroke. The new model of distal middle cerebral artery occlusion is a stable, reliable model, with a clear infarct range, which is suitable for the study of ischemic stroke.


Key words: stroke, ischemic stroke, middle cerebral artery, distal occlusion, pathological changes, behavior, model, mouse

中图分类号: