法医学杂志

法医学杂志 ›› 2019, Vol. 35 ›› Issue (2): 136-142.DOI: 10.12116/j.issn.1004-5619.2019.02.002

所属专题： 损伤时间推断

• 专题 • 上一篇    下一篇

小鼠脑挫伤后CB2R的表达变化与损伤时间的关系

陈京伟1，2， 王鹏飞1，2， 张孟周1，2， 张忠铎1，2， 程浩1，2， 孙英富1，2， 温书恒1，2， 郭相伸1，2， 赵锐1，2， 官大威1，2   

  1. 1. 中国医科大学法医学院 法医司法鉴定中心，辽宁 沈阳 110122； 2. 智慧检务创新研究院 智慧司法鉴定联合实验室，辽宁 沈阳 110122
  • 发布日期:2019-04-25 出版日期:2019-04-28
  • 通讯作者: 官大威，男，博士，教授，博士研究生导师，主要从事器官组织损伤修复分子生物学机制与损伤时间推断研究；E-mail：dwguan@cmu.edu.cn
  • 作者简介:陈京伟（1990—），男，硕士研究生，主要从事颅脑损伤机制及损伤时间推断研究；E-mail：1334789226@qq.com
  • 基金资助:
    国家自然科学基金资助项目（81671862，81772023，81801874，81871529）；国家重点研发项目子课题资助项目（2018YFC 0807204）

Relationship between Expression Changes of CB2R and Wound Age of Brain Contusion in Mice

CHEN Jing-wei1,2, WANG Peng-fei1,2, ZHANG Meng-zhou1,2, ZHANG Zhong-duo1,2, CHENG Hao1,2, SUN Ying-fu1,2, WEN Shu-heng1,2, GUO Xiang-shen1,2, ZHAO Rui1,2, GUAN Da-wei1,2   

  1. 1. Center of Medico-legal Investigation, School of Forensic Medicine, China Medical University, Shenyang 110122, China; 2. Collaborative Laboratory of Intelligentized Forensic Science, Shenyang 110122, China
  • Online:2019-04-25 Published:2019-04-28

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摘要： 目的 检测小鼠脑挫伤后不同时间点大麻素2型受体（cannabinoid type 2 receptor，CB2R）的表达情况，并探索其与损伤时间的相关性。 方法 通过PCI3000精密颅脑损伤撞击器建立小鼠脑挫伤模型，利用免疫组织化学染色、免疫荧光染色和Western印迹法检测不同损伤时间点损伤区周围CB2R的表达变化。 结果 免疫组织化学染色结果显示，假手术组脑皮质中仅少量细胞呈CB2R阳性表达，脑挫伤后CB2R阳性细胞率逐渐升高，于伤后12 h和伤后7 d两次达到高峰，随后逐渐下降，至伤后28 d基本恢复至正常水平。Western印迹法检测结果与免疫组织化学染色结果一致。免疫荧光染色结果显示，脑挫伤后CB2R阳性神经元、CB2R阳性单核细胞、CB2R阳性星形胶质细胞数与总细胞数的比值变化均呈单峰模式，分别于伤后12 h、1 d、7 d达到高峰。 结论 小鼠脑挫伤后CB2R在神经元、单核细胞及星形胶质细胞中的表达提示，其可能参与调节这些细胞的生物学功能。脑挫伤后CB2R的动态表达存在时间规律性，有望成为法医学脑挫伤损伤时间推断的生物学指标。

关键词: 法医病理学, 脑损伤, 免疫组织化学染色, 免疫荧光技术, 印迹法, 蛋白质, 大麻素2型受体, 损伤时间推断, 小鼠

Abstract: Objective To investigate the expression of cannabinoid type 2 receptor （CB2R） at different time points after brain contusion and its relationship with wound age of mice. Methods A mouse brain contusion model was established with PCI3000 Precision Cortical Impactor. Expression changes of CB2R around the injured area were detected with immunohistochemical staining, immunofluorescent staining and Western blotting at different time points. Results Immunohistochemical staining results showed that only a few cells in the cerebral cortex of the sham operated group had CB2R positive expression. The ratio of CB2R positive cells gradually increased after injury and reached the peak twice at 12 h and 7 d post-injury, followed by a decrease to the normal level 28 d post-injury. The results of Western blotting were consistent with the immunohistochemical staining results. Immunofluorescent staining demonstrated that the changes of the ratio of CB2R positive cells in neurons, CB2R positive cells in monocytes and CB2R positive cells in astrocytes to the total cell number showed a single peak pattern, which peaked at 12 h, 1 d and 7 d post-injury, respectively. Conclusion The expression of CB2R after brain contusion in neurons, monocytes and astrocytes in mice suggests that it is likely to be involved in the regulation of the biological functions of those cells. The changes in CB2R are time-dependent, which suggests its potential applicability as a biological indicator for wound age estimation of brain contusion in forensic practice.

Key words: forensic pathology, brain injuries, immunohistochemistry staining, immunofluorescence technique, blotting, Western, cannabinoid type 2 receptor, wound age estimation, mice