轻中度闭合型创伤性脑损伤小鼠模型建立及评价  

Establishment and evaluation of a mild-to-moderate closed traumatic brain injury mouse model

作　　者：卜苑静 付文亮 邢微微 徐东刚 BU Yuanjing;FU Wenliang;XING Weiwei;XU Donggang(School of Basic Medicine,Anhui Medical University,Hefei 230032,China;Institute of Military Cognition and Brain Sciences,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850)

机构地区：[1]安徽医科大学基础医学院,合肥230032 [2]军事科学院军事医学研究院军事认知与脑科学研究所,北京100850

出　　处：《中国实验动物学报》2023年第9期1140-1147,共8页Acta Laboratorium Animalis Scientia Sinica

摘　　要：目的 建立稳定的轻中度闭合型创伤性脑损伤小鼠模型并对其进行评价。方法 将170只SPF级ICR小鼠分为假手术组(Sham组,n=34)和模型组(TBI组,n=136),模型组分为TBI-A组、TBI-B组、TBI-C组和TBI-D组,其致伤高度均为20 cm,下落砝码重量分别为60 g、80 g、100 g和120 g。按照以上条件对小鼠进行脑损伤敲击,每组34只。造模后通过mNSS评分、疲劳转棒实验、Morris水迷宫实验评估小鼠平衡运动能力和学习记忆能力;脑含水量实验、伊文思蓝实验、苏木精-伊红(HE)染色、GFAP免疫组化以及TUNEL免疫荧光分析脑组织病理改变。结果 与Sham组相比,TBI组的mNSS评分值和水迷宫逃避潜伏期出现不同程度的升高,持续在转棒上行走的时间明显下降,表明TBI组小鼠出现严重的神经功能障碍。TBI组小鼠脑组织内的含水量和伊文思蓝含量明显高于Sham组,提示该组小鼠血脑屏障受损并出现脑水肿。组织病理学检测显示,TBI组小鼠脑组织神经元出现明显变性皱缩,星形胶质细胞显著增生且凋亡细胞比例显著增加,损伤程度随打击物重量的增加而加重。综合分析表明,在下落高度为20 cm,打击重量为60 g和80 g时表现轻度颅脑损伤,打击重量为100 g和120 g时可模拟出为中度颅脑损伤。结论 基于自由落体原理建立了稳定的轻中度闭合型小鼠脑损伤模型,为轻中度创伤性脑损伤的机制和治疗研究奠定了基础。Objective To establish a stable mouse model of mild-to-moderate closed head injury and evaluate its effectiveness.Methods A total of 170 SPF grade ICR mice were divided into a sham group(n=34)and TBI group(n=136).The TBI group was further divided into four subgroups,TBI-A,TBI-B,TBI-C,and TBI-D,with a drop height of 20 cm and drop weights of 60 g,80 g,100 g,and 120 g,respectively.According to the above conditions,the brain injury percussion was performed on the mice,34 mice in each group.After modeling,the mNSS,fatigue rotating test,and Morris water maze test were used to assess balance and learning abilities of the mice.Brain water content analysis,evens blue staining,HE staining,GFAP immunohistochemistry,and TUNEL immunofluorescence were used to analyze pathological changes in brain tissue.Results Compared with the sham group,the mNSS and water maze escape latency in the TBI group increased to various degrees,whereas continuous walking time on the ratating rod decreased significantly,indicating severe neurological dysfunction in the TBI group.Brain water content and evens blue staining in brain tissue of TBI group mice were significantly higher than those in the sham group,suggesting blood-brain barrier damage and brain edema.Histopathological examination showed that neurons in brain tissue of the TBI group showed obvious degeneration and contraction,astrocyte proliferation,and the proportion of apoptotic cells was increased significantly.The damage degree increased with the increase in the weight of the impact.Comprehensive analysis revealed that weights of 60 g or 80 g with a height of injury at 20 cm simulated mild closed head injury in mice,whereas weights of 100 g or 120 g simulated moderate closed head injury in mice.Conclusions A stable mouse model of mild-to-moderate closed head injury was established using the principle of free fall,which lays the foundation to study the mechanism and treatment strategies for mild-to-moderate traumatic brain injury.

关 键 词：自由落体撞击 组织学分析 轻中度颅脑损伤 小鼠模型 神经功能障碍 

分 类 号：Q95-33[生物学—动物学]