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电针对不同剂量X射线照射C57小鼠神经干细胞增殖和分化的影响及机制

武鑫, 苏少华, 孙宁宁, 吕明惠, 张松江, 高剑峰^*

河南中医药大学基础医学院，郑州 450046

摘要

本文旨在探讨电针对不同剂量X射线照射C57小鼠海马区内源性神经干细胞增殖、分化及Notch信号通路的影响。将30日龄C57BL/6J小鼠随机分为对照组、放射线照射组和电针组。对照组不接受照射处理，放射线照射组小鼠接受10 min的4、8或16 Gy X射线照射，电针组在相应照射后接受3个疗程的电针(百会、风府和双侧肾俞)治疗。用免疫组织化学方法检测海马区内源性神经干细胞增殖和分化情况，用RT-PCR和Western blot分别检测海马区Notch信号通路相关基因mRNA和蛋白的表达。结果显示，与对照组比较，放射线照射组海马BrdU阳性细胞(4、8 Gy亚组)和BrdU/NeuN双标阳性细胞(3个剂量亚组)数目均显著减少，而电针治疗可逆转以上变化。放射线照射组各剂量亚组BrdU/GFAP双标阳性细胞数目相对对照组均显著减少，而电针治疗可以逆转4和8 Gy剂量亚组的这一变化。此外，与对照组比较，放射线照射组各剂量亚组小鼠海马Notch1 mRNA及蛋白表达均显著上调，而Mash1基因及蛋白表达显著下降；而与放射线照射组比较，电针组各剂量亚组海马Notch1 mRNA及蛋白表达均显著下降，4和8 Gy亚组Mash1 mRNA和蛋白表达均显著增加。上述结果提示，放射线照射可以抑制小鼠海马区神经干细胞增殖和分化，电针(百会、风府和双侧肾俞)能够显著提高放射线照射小鼠海马区神经干细胞的增殖和分化，这一作用可能与Notch蛋白信号通路相关。

关键词： 放射线照射; 电针; 神经干细胞; Notch信号通路

分类号：R338

Effects and mechanisms of electro-acupuncture on proliferation and differentiation of neural stem cells in C57 mice exposed to different doses of X-ray radiation

WU Xin, SU Shao-Hua, SUN Ning-Ning, LYU Ming-Hui, ZHANG Song-Jiang, GAO Jian-Feng^*

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

Abstract

The present study was aimed to investigate the effects and mechanisms of electro-acupuncture (EA) on proliferation and differentiation of neural stem cells in the hippocampus of C57 mice exposed to different doses of X-ray radiation. Thirty-day-old C57BL/6J mice were randomly divided into control, irradiation, and EA groups. The control group was not treated with irradiation. The irradiation groups were exposed to different doses of X-ray (4, 8 or 16 Gy) for 10 min. The EA groups were electro-acupunctured at Baihui, Fengfu and bilateral Shenyu for 3 courses of treatment after X-ray radiation. Immunohistochemistry was used to evaluate proliferation and differentiation of the hippocampal neural stem cell. RT-PCR and Western blot were used to detect mRNA and protein expressions of Notch1 and Mash1 in the hippocampus, respectively. The results showed that, compared with the control group, the numbers of BrdU positive cells (4, 8 Gy subgroup) and BrdU/NeuN double-labeling positive cells (3 dose subgroups) were decreased significantly in the irradiation group, but the above changes could be reversed by EA. Compared with the control group, the number of BrdU/GFAP double-labeling positive cells in each dose subgroup of irradiation group was decreased significantly, while EA could reverse the change of 4 and 8 Gy dose subgroups. In addition, compared with the control group, the expression levels of Notch1 mRNA and protein in hippocampus were up-regulated, and the expression levels of Mash1 mRNA and protein were significantly decreased in each dose subgroup of irradiation group. Compared with irradiation group, the expression levels of Notch1 mRNA and protein in hippocampus of EA group were decreased significantly in each dose subgroup, and the expression levels of Mash1 mRNA and protein were increased significantly in 4 and 8 Gy subgroups. These results suggest that irradiation affects the proliferation and differentiation of neural stem cells in hippocampus of mice, whereas EA may significantly increase the proliferation and differentiation of hippocampal neural stem cells via the regulation of Notch signaling pathway. 

Key words: irradiation; electro-acupuncture; neural stem cells; Notch signaling pathway

收稿日期：2018-07-03　　录用日期：2019-01-11

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

DOI: 10.13294/j.aps.2019.0036

引用本文：

武鑫, 苏少华, 孙宁宁, 吕明惠, 张松江, 高剑峰. 电针对不同剂量X射线照射C57小鼠神经干细胞增殖和分化的影响及机制[J]. 生理学报 2019; 71 (3): 431-438.

WU Xin, SU Shao-Hua, SUN Ning-Ning, LYU Ming-Hui, ZHANG Song-Jiang, GAO Jian-Feng. Effects and mechanisms of electro-acupuncture on proliferation and differentiation of neural stem cells in C57 mice exposed to different doses of X-ray radiation. Acta Physiol Sin 2019; 71 (3): 431-438 (in Chinese with English abstract).