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中华脑科疾病与康复杂志(电子版) ›› 2021, Vol. 11 ›› Issue (04) : 215 -220. doi: 10.3877/cma.j.issn.2095-123X.2021.04.006

基础研究

CR2-Crry预处理诱导神经干细胞在颅脑损伤中发挥神经保护作用
高谋1, 徐如祥2,(), 董勤3, 郭莉丽2   
  1. 1. 100853 北京,解放军总医院第一医学中心神经外科医学部
    2. 610072 成都,四川省医学科学院·四川省人民医院神经外科
    3. 100038 北京,首都医科大学附属复兴医院神经内科
  • 收稿日期:2021-05-13 出版日期:2021-08-15
  • 通信作者: 徐如祥
  • 基金资助:
    国家自然科学基金(81671189、81971295、82001293); 全军"十三五"军事医学创新工程(16CXZ001)

Transplantation of induced neural stem cells receiving CR2-Crry pre-treatment mediates neuroprotection following brain injury

Mou Gao1, Ruxiang Xu2,(), Qin Dong3, Lili Guo2   

  1. 1. Department of Neurosurgery, The First Medical Center of PLA General Hospital, Beijing 100853, China
    2. Department of Neurosurgery, Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, Chengdu 610072, China
    3. Department of Neurology, Fu Xing Hospital, Capital Medical University, Beijing 100038, China
  • Received:2021-05-13 Published:2021-08-15
  • Corresponding author: Ruxiang Xu
引用本文:

高谋, 徐如祥, 董勤, 郭莉丽. CR2-Crry预处理诱导神经干细胞在颅脑损伤中发挥神经保护作用[J]. 中华脑科疾病与康复杂志(电子版), 2021, 11(04): 215-220.

Mou Gao, Ruxiang Xu, Qin Dong, Lili Guo. Transplantation of induced neural stem cells receiving CR2-Crry pre-treatment mediates neuroprotection following brain injury[J]. Chinese Journal of Brain Diseases and Rehabilitation(Electronic Edition), 2021, 11(04): 215-220.

目的

探讨CR2-Crry预处理诱导神经干细胞(iNSCs)对颅脑损伤的神经保护作用。

方法

18只雄性成年C57BL/6小鼠制备颅脑损伤模型,神经功能缺损评分(NSS)4~8分的小鼠为颅脑损伤组,按照随机数字表法分为:CR2-Crry预处理iNSCs(CR2-Crry-iNSCs)组(6只)、磷酸盐缓冲液(PBS)预处理iNSCs(PBS-iNSCs)组(6只)和PBS组(6只)。于颅脑损伤后12 h分别将1×106个CR2-Crry-iNSCs、PBS-iNSCs或等体积PBS经立体定向移植到颅脑损伤小鼠脑内。于颅脑损伤后14 d处死小鼠,制备脑组织冰冻切片,并行双重免疫荧光和原位末端标记(TUNEL)染色,观察CR2-Crry-iNSCs和PBS-iNSCs对颅脑损伤小鼠脑内Crry、NeuN和TUNEL阳性细胞数量的影响。

结果

双重免疫荧光染色:与PBS组比较,PBS-iNSCs组和CR2-Crry-iNSCs组颅脑损伤小鼠脑内Crry和NeuN双阳性的神经细胞数量明显增加,差异具有统计学意义(P<0.05)。此外,与PBS-iNSCs组比较,CR2-Crry-iNSCs组颅脑损伤小鼠脑内Crry和NeuN双阳性的神经细胞数量明显增加,差异具有统计学意义(P<0.05)。TUNEL染色:与PBS组比较,PBS-iNSCs组和CR2-Crry-iNSCs组颅脑损伤小鼠脑内TUNEL和NeuN双阳性的神经细胞数量明显下降,差异具有统计学意义(P<0.05)。此外,与PBS-iNSCs组比较,CR2-Crry-iNSCs组颅脑损伤小鼠脑内TUNEL和NeuN双阳性的神经细胞数量明显下降,差异具有统计学意义(P<0.05)。

结论

CR2-Crry预处理iNSCs移植可增加颅脑损伤小鼠脑内神经细胞Crry表达,减轻补体介导的神经损伤。

Objective

To study the effects of transplanted induced neural stem cells (iNSCs) receiving CR2-Crry pre-treatment on neuroprotection following brain injury.

Methods

Healthy adult male C57BL/6 mouse brain injury models were established and mice having an neurological severity scores (NSS) of 4-8 were enrolled in brain injury group (n=18). These brain injury animals were randomly divided into 3 groups: the CR2-Crry-iNSC group (n=6), the phosphate buffered solution (PBS)-iNSC group (n=6) and the PBS group (n=6). At 12 h after brain injury, CR2-Crry-iNSCs, PBS-iNSCs or PBS were separately injected into the brains of brain injury mice via stereotactic method. On day 14 after brain injury, mice were sacrificed for morphological analysis. Immunofluorescent staining and TdT-mediated dUTP nick and labeling (TUNEL) staining were utilized to determine the effects of CR2-Crry-iNSC and PBS-iNSC grafts on Crry+/NeuN+ and NeuN+/TUNEL+ cells in the brains of brain injury mice.

Results

Double-labeling experiments revealed that the level of Crry+/NeuN+ neurons in the brains of the PBS group was significantly lower than that in CR2-Crry-iNSC and PBS-iNSC groups (P<0.05). Furthermore, the level of Crry+/NeuN+ neurons was substantially higher in the CR2-Crry-iNSC group than in the PBS-iNSC group (P<0.05). In contrast, TUNEL staining showed that the level of NeuN+/TUNEL+ neurons in the brains of the PBS group was significantly higher than that in CR2-Crry-iNSC and PBS-iNSC groups (P<0.05). Moreover, the level of NeuN+/TUNEL+ neurons was substantially lower in the CR2-Crry-iNSC group than in the PBS-iNSC group (P<0.05).

Conclusion

Transplantation of iNSCs receiving CR2-Crry pre-treatment could increase the levels of Crry expression in neurons and alleviate complement-mediated neuronal damage following brain injury.

图1 颅脑损伤14 d后的各组小鼠脑内Crry和NeuN双阳性细胞的分布
表1 颅脑损伤14 d后的3组小鼠脑内Crry、NeuN和TUNEL阳性细胞统计(Mean±SD,%)
图2 颅脑损伤14 d后的各组小鼠脑内TUNEL和NeuN双阳性细胞的分布
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