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不同疗程电针对放射性脑损伤小鼠突触超微结构及突触功能相关蛋白的影响

武鑫, 李炎辉, 张文靖, 张松江, 高剑峰^*

河南中医药大学基础医学院，郑州 450046

摘要

本研究旨在探讨不同疗程电针穴位干预对放射线照射所致脑损伤模型小鼠突触结构及突触功能相关蛋白表达的影响。60只1月龄C57BL/6J雄性小鼠随机分为对照组、放射性脑损伤模型组、电针1周组、电针2周组、电针3周组和电针对照组。模型组小鼠接受X-射线照射(8 Gy, 10 min)建立放射性脑损伤模型，电针组在X-射线照射后分别接受1周、2周和3周电针穴位(百会、风府、双侧肾俞)干预，电针对照组在照射后进行非穴位电针干预。免疫组织化学染色检测海马CA1区突触超微结构，RT-qPCR和Western blot检测脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)、突触蛋白-1 (synapsin-1)和突触后致密蛋白95 (postsynaptic density 95, PSD95)的基因和蛋白表达。结果显示，与对照组比较，模型组和电针对照组小鼠神经元核间隙明显减小，神经元核质比显著增大，突触间隙和突触后致密物(postsynaptic density, PSD)厚度则明显降低，线粒体表面积密度、体密度和比表面均显著降低。与模型组比较，电针2周组小鼠的神经元核质比显著减小，PSD厚度显著增加，线粒体体密度显著增加；电针3周组小鼠核间隙显著增加，核质比显著减小，突触间隙和PSD厚度均显著增大，线粒体表面积密度和比表面均显著增加。此外，与对照组比较，模型组和电针对照组小鼠海马CA1区BDNF、synapsin-1、PSD95基因及蛋白表达均不同程度地下降。而与模型组比较，电针各组synapsin-1基因表达均显著上调，电针1和2周组BDNF基因表达显著上调，电针2周组PSD95基因表达显著上调，而电针各组3种蛋白表达均显著上调。 以上结果表明，放射线照射损伤小鼠海马CA1区突触超微结构及突触功能相关蛋白表达，电针干预可显著改善放射线照射所导致的突触结构和功能损伤。

关键词： 电针; 海马区; 突触结构; 突触可塑性

分类号：R338

Effects of electroacupuncture with different courses on the synaptic structure and synaptic function-related proteins in mice with radiation-induced brain injury 

WU Xin, LI Yan-Hui, ZHANG Wen-Jing, ZHANG Song-Jiang, GAO Jian-Feng^*

School of Basic Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, China

Abstract

The aim of the present study was to investigate the effects of different courses of electroacupuncture on synaptic structure and synaptic function-related proteins expression in the hippocampal CA1 region of radiation-induced brain injury mice. Sixty C57BL/6J male mice were randomly divided into control group, radiation-induced brain injury model group, 1-week electroacupuncture group (EA1), 2-week electroacupuncture group (EA2), 3-week electroacupuncture group (EA3), and electroacupuncture-control (EA-Ctrl) group. The mice in model group were exposed to X-ray irradiation (8 Gy, 10 min) to establish radiation-induced brain injury model. The mice in EA groups were acupunctured at electroacupuncture points (Baihui, Fengfu and bilateral Shenshu) for 1 week, 2 weeks and 3 weeks respectively after radiation. Immunohistochemistry was used to observe synaptic structure in hippocampal CA1 region. The expressions of brain-derived neurotrophic factor (BDNF), synapsin-1 and postsynaptic density 95 (PSD95) in the hippocampal CA1 region of each group were detected by RT-PCR and Western blotting. The results showed that the nuclear gap in model and EA-Ctrl groups was significantly decreased compared to control group, however nucleus to cytoplasm ratio was significantly increased. The synaptic cleft, postsynaptic density (PSD) thickness, the mitochondrial surface density, volume density and specific surface area were significantly reduced. Compared with model group, the nucleus to cytoplasm ratio of EA2 group was significantly decreased, the PSD thickness and mitochondrial volume density were significantly increased; the nuclear gap of EA3 group was significantly increased, nucleus to cytoplasm ratio was significantly decreased, synaptic cleft and PSD thickness were significantly increased, and the mitochondrial surface density and specific surface area were all increased significantly. In addition, compared with the control group, the gene and protein expressions of BDNF, synapsin-1 and PSD95 in the hippocampal CA1 region of the model group and EA-Ctrl group were significantly decreased. However, compared with the model group, the gene expression of synapsin-1 in EA groups was significantly up-regulated, the gene expression of BDNF in EA1 and EA2 groups was significantly up-regulated, and the gene expression of PSD95 in EA2 group was significantly up-regulated. Moreover, the protein expressions of BDNF, synapsin-1 and PSD95 of EA groups were significantly up-regulated compared with the model group. These results indicate that the synaptic structure and the expression of synaptic function-related proteins in hippocampal CA1 region were injured by radiation exposure, whereas electroacupuncture intervention can significantly improve the synaptic structure and function damage caused by radiation. 

Key words: Electroacupuncture; hippocampus;;

收稿日期：2021-01-16　　录用日期：2021-05-13

通讯作者：高剑峰　　E-mail: jc.gjfeng@hotmail.com

引用本文：

武鑫, 李炎辉, 张文靖, 张松江, 高剑峰. 不同疗程电针对放射性脑损伤小鼠突触超微结构及突触功能相关蛋白的影响[J]. 生理学报 2021; 73 (6): 909-916.

WU Xin, LI Yan-Hui, ZHANG Wen-Jing, ZHANG Song-Jiang, GAO Jian-Feng. Effects of electroacupuncture with different courses on the synaptic structure and synaptic function-related proteins in mice with radiation-induced brain injury . Acta Physiol Sin 2021; 73 (6): 909-916 (in Chinese with English abstract).