Effects of different courses of electrotherapy on learning, memory, and Notch signaling in a mouse model of radiation-induced brain injury

LI Yanhui; WU Xin; WANG Tan; WANG Baiyan; ZHANG Han; NIU Tingyuan; NIU Yuqi; GAO Jianfeng

doi:10.11889/j.1000-3436.2023-0004

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Effects of different courses of electrotherapy on learning, memory, and Notch signaling in a mouse model of radiation-induced brain injury

RADIOBIOLOGY AND RADIOMEDICINE | 更新时间：2023-10-27

- Effects of different courses of electrotherapy on learning, memory, and Notch signaling in a mouse model of radiation-induced brain injury
  封面论文
- Journal of Radiation Research and Radiation Processing Vol. 41, Issue 5, Pages: 50301(2023)
- 作者机构：
  
  1.河南中医药大学医学院郑州 450046
- 作者简介：
  
  LI Yanhui (male) was born in December 1996, Now he is a master student, research direction: prevention and treatment of brain injury with traditional Chinese medicine, E-mail: 651148324@qq.com
  GAO Jianfeng, medical doctor, professor, master's supervisor, E-mail: jc.gjfeng@hotmail.com
- 基金信息：
  
  Natural Science Foundation of Henan Province(202300410267);Key R&D and Promotion Project of Henan Province(212102311082);The Young Key Teachers Training Program of Henan Province(2021GGJS085);2021 Postgraduate Research Innovation Project of Henan University of Traditional Chinese Medicine(2021KYCX043);2019 Graduate Innovation and Entrepreneurship Team of Henan University of Chinese Medicine(2019CXCY001)
- DOI：10.11889/j.1000-3436.2023-0004
  CLC：
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LI Yanhui, WU Xin, WANG Tan, et al. Effects of different courses of electrotherapy on learning, memory, and Notch signaling in a mouse model of radiation-induced brain injury. [J]. Journal of Radiation Research and Radiation Processing 41(5):050301(2023)
DOI：

LI Yanhui, WU Xin, WANG Tan, et al. Effects of different courses of electrotherapy on learning, memory, and Notch signaling in a mouse model of radiation-induced brain injury. [J]. Journal of Radiation Research and Radiation Processing 41(5):050301(2023) DOI： 10.11889/j.1000-3436.2023-0004.

摘要

探讨不同疗程的电针干预对放射性脑损伤小鼠学习记忆的影响及其作用机制。60只4周龄雄性C57/BL6J小鼠随机分为空白组、模型组、电针组1（7 d）、电针组2（14 d）、电针组3（21 d）、电针组4（28 d）。除空白组外，其余各组给予X射线（8 Gy，2 min）照射，构建放射性脑损伤模型，电针组给予针刺“百会”、“风府”及双侧“肾俞”穴，并依据分组分别干预7、14、21、28 d。电针结束后，Y迷宫检测各组小鼠学习记忆功能，苏木精-伊红染色法、透射电镜观察海马齿状回（DG）神经元形态、突触超微结构，Western blot检测Notch信号通路Notch 1、Hes 1、ASCL 1的蛋白表达。结果显示：与空白组相比，模型组小鼠学习记忆功能显著下降（,p,<,0.01）；海马DG区神经元数量减少，排列紊乱；神经元突触前膜突触囊泡数量减少，突触界面率减小、突触后致密区（PSD）厚度降低、突触间隙增大均有显著差异（,p<,0.01）；与模型组相比，电针各组小鼠的学习记忆功能均显著提高（,p<,0.05，,p<,0.01）；电针组小鼠DG区神经元数量增多，排列整齐；各组小鼠DG区突触界面率均有显著改善（,p<,0.01），电针组2、3和4小鼠PSD厚度显著增加（,p<,0.01），电针组2、3和4的突触间隙显著减小（,p<,0.01）。与空白组相比，模型组小鼠Notch 1蛋白表达显著升高（,p,<,0.05），Hes 1蛋白表达显著升高（,p,<,0.01），ASCL 1蛋白表达显著降低（,p,<,0.01）；与模型组相比，电针组2、3和4的Notch 1、Hes 1蛋白表达显著降低（,p<,0.05，,p,<,0.01），电针组3和4的ASCL 1蛋白表达显著升高（,p<,0.05，,p,<,0.01）。提示不同疗程的电针可以改善放射线照射对小鼠学习记忆功能造成的损伤，该作用与改善DG区突触超微结构及调控Notch信号通路相关蛋白的表达有关。

Abstract

Herein, we investigated the effects of different courses of electroacupuncture on learning and memory in mice with radiation-induced brain injury, and elucidated the underlying mechanisms. Sixty 4-week-old male C57/BL6J mice were randomly divided into the blank group, model group, electroacupuncture group 1 (7 d), electroacupuncture group 2 (14 d), electroacupuncture group 3 (21 d), and electroacupuncture group 4 (28 d). Except the blank group, all groups were irradiated with X-ray (8 Gy, 2 min) to construct the radioactive brain injury model. The electroacupuncture group was injected with "Baihui," "Fengfu," and bilateral "Shenshu" points, and the intervention was performed for 7, 14, 21 and 28 days according to the groups, respectively. After electroacupuncture, the Y-maze test was conducted to assess the learning and memory of mice in each group. Neuron morphology and synaptic ultrastructure in the hippocampal DG region were observed via Hematoxylin-eosin staining and transmission electron microscopy. Protein expression levels of the Notch signaling pathway components Notch 1, Hes 1, and ASCL 1 were detected via Western blotting. The results revealed that, compared to those in the blank group, the learning and memory function of mice decreased significantly in the model group (,p,<,0.01). The number of neurons in the hippocampal dentategyrus (DG) region decreased and their arrangement was disordered. The number of synaptic vesicles in the presynaptic membrane decreased, the rate of synaptic interface decreased, the thickness of the postsynaptic density (PSD) decreased, and the synaptic cleft increased significantly (,p,<,0.01). Compared to those of the model group, the learning and memory function of mice in electroacupuncture group was significantly improved (,p,<,0.05,p,<,0.01). In the electroacupuncture group, the number of neurons in the DG region increased, with neurons neatly arranged. The rate of synaptic interface in the DG region improved significantly in all groups (,p,<,0.01), the PSD thickness of mice in groups 2, 3, and 4 was significantly increased (,p,<,0.01), and the synaptic cleft in groups 2, 3, and 4 was significantly decreased (,p,<,0.01). Compared to the blank group, Notch 1 (,p,<,0.05) and Hes 1 (,p,<,0.01) protein expressions were significantly increased in the model group, while ASCL 1 protein expression was significantly decreased (,p,<,0.01); compared those in the model group, the expressions of Notch 1 and Hes 1 proteins in electroacupuncture groups 2, 3, and 4 were significantly decreased (,p,<,0.05, p,<,0.01), and the expression of ASCL 1 protein was significantly increased in electroacupuncture groups 3 and 4 (,p,<,0.05 and ,p,<,0.01, respectively ). In conclusion, different courses of electroacupuncture can ameliorate the damage in learning and memory function caused by irradiation in mice, which is associated with improving the ultrastructure of synapses in the DG region and regulating the expression of related proteins of the Notch signaling pathway.

关键词

放射性脑损伤电针学习记忆Notch信号通路

Keywords

Radiation-induced brain injuryElectropunctureLearning and memoryNotch signaling pathway

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