中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (19): 3009-3015.doi: 10.3969/j.issn.2095-4344.3538

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

放散式冲击波通过Notch1/Hes1通路调节脑缺血后海马组织中神经干细胞的增殖与分化

苏明珠1，马跃文1，2   

  1. 1中国医科大学附属第一医院，辽宁省沈阳市  110001；2中国医科大学附属第一医院康复医学科，辽宁省沈阳市  110001
  • 收稿日期:2020-05-07 修回日期:2020-05-13 接受日期:2020-07-17 出版日期:2021-07-09 发布日期:2021-01-13
  • 通讯作者: 马跃文，博士，教授，主任医师，中国医科大学附属第一医院康复医学科，辽宁省沈阳市 110001
  • 作者简介:苏明珠，女，1993年生，湖南省平江县人，汉族，2019年中国医科大学毕业，硕士，医师，主要从事神经康复方面的研究。
  • 基金资助:
    辽宁省重点研发计划指导计划项目(2017225013)，项目负责人：马跃文

Radial extracorporeal shock wave therapy regulates the proliferation and differentiation of neural stem cells in the hippocampus via Notch1/Hes1 pathway after cerebral ischemia

Su Mingzhu1, Ma Yuewen1, 2   

  1. 1The First Hospital of China Medical University, Shenyang 110001, Liaoning Province, China; 2Department of Rehabilitation Medicine, The First Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
  • Received:2020-05-07 Revised:2020-05-13 Accepted:2020-07-17 Online:2021-07-09 Published:2021-01-13
  • Contact: Ma Yuewen, MD, Professor, Chief physician, The First Hospital of China Medical University, Shenyang 110001, Liaoning Province, China; Department of Rehabilitation Medicine, The First Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
  • About author:Su Mingzhu, Master, Physician, The First Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
  • Supported by:
    the Liaoning Provincial Key Research & Development Guidance Program, No. 2017225013 (to MYW)

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摘要：

文题释义：
冲击波：冲击波治疗仪是以压缩空气驱使子弹体撞击治疗探头产生弹道式冲击波应用于机体来治疗疾病。冲击波作用于意识障碍患者的额叶，可以促进患者苏醒；作用于人头部三叉神经节体面投影处可以有效缓解三叉神经痛；作用于脑缺血大鼠头部，可以降低炎症反应，减少缺血体积。
Notch1信号通路：Notch信号通路在哺乳动物中有4个同源受体，分别为 Notch1-Notch4，Notch配体有Dill、Jagged1等。脑缺血后，Notch1和其配体Jagged1在神经干细胞的增殖与分化过程中作用更为明显。激活Notch1信号通路能诱导神经干细胞表现出更大的分化潜能，修复缺血后神经元损伤。NICD是Notch信号通路的胞内活化因子，脑缺血后室管膜下区NICD水平明显升高，可促进神经干细胞增殖和迁移。Hes1是Notch1信号通路下游的一个关键基因，在维持神经干细胞未分化状态中有重要作用。脑缺血后，Hes1 受上游信号和细胞因子的共同调控，影响神经干细胞增殖和分化，发挥修复作用。 

背景：作者前期研究发现，放散式冲击波在体外可以促进神经干细胞增殖和向神经元分化，其具体机制与上调Notch1信号通路有关，但在体内是否也可以通过该途径促进神经干细胞增殖和分化，目前尚未见相关研究报道。
目的：探究放散式冲击波在体内是否可以通过Notch1/Hes1信号通路影响脑缺血后大鼠海马组织中神经干细胞的增殖与分化。
方法：采用大脑中动脉栓塞法建立大鼠右侧脑缺血模型，造模成功后随机分成2组：治疗组和对照组。治疗组：大脑中动脉栓塞后72 h采用放散式冲击波直接作用于大鼠右侧头部，治疗剂量：表面探头，0.1 MPa，10 Hz，200点，每3 d治疗1次；对照组：大脑中动脉栓塞后72 h，给予同样剂量的放散式冲击波，但探头不接触大鼠头部，只有声音的刺激。大脑中动脉栓塞后第12，21，30天通过mNSS评分观察神经功能的恢复情况，通过免疫组化、Western blot、RT-PCR检测大鼠缺血侧海马组织中Nestin、NSE、Notch1和Hes1的表达。
结果与结论：①治疗组在第12，21，30天的mNSS评分明显低于对照组，说明放散式冲击波可以明显改善大鼠脑缺血后的神经功能障碍；②与对照组相比，治疗组大鼠缺血侧海马组织内Nestin和NSE的表达明显增高，提示放散式冲击波能促进脑缺血后海马组织中神经干细胞的增殖与分化；③与对照组相比，治疗组大鼠缺血侧海马组织内Notch1和Hes1的表达增高，提示放散式冲击波可以上调海马区Notch1/ Hes1信号通路的表达；④结果表明，放散式冲击波可以促进脑缺血后海马区神经干细胞的增殖与分化，从而改善神经功能，其具体机制与上调Notch1/Hes1信号通路有关。

关键词: 干细胞, 神经干细胞, 放散式冲击波, Notch1, 信号通路, 海马, 大鼠

Abstract: BACKGROUND: Our previous research found that radial extracorporeal shock wave therapy can promote the proliferation and differentiation of neural stem cells in vitro, and the specific mechanism is related to the upregulation of Notch1 signaling pathway. However, there is no relevant research report on whether the proliferation and differentiation of neural stem cells can also be promoted through this pathway in vivo.
OBJECTIVE: To explore whether radial extracorporeal shock wave therapy can promote the proliferation and differentiation of neural stem cells in hippocampus after cerebral ischemia through Notch1/Hes1 signaling pathway in vivo. 
METHODS:  Rat models of right cerebral ischemia were established by middle cerebral artery occlusion. After successful modeling, the rats were randomly divided into two groups: the radial extracorporeal shock wave therapy group (treatment group) and the control group. In the treatment group, 72 hours after middle cerebral artery occlusion, radial extracorporeal shock wave therapy was directly applied to the right side of the rats’ head at a dose of 0.1 MPa, 200 impulses, and 10 Hz, every three days. In the control group, 72 hours after middle cerebral artery occlusion, the same dose of radial extracorporeal shock wave therapy was given, but the transmitter probe did not touch the head of the rats, only the sound stimulation. Both groups were observed at 12, 21 and 30 days after middle cerebral artery occlusion. Expression levels of nestin, NSE, Notch1 and Hes1 in the ischemic hippocampus of rats were detected by immunohistochemistry, western blot assay, and RT-PCR.
RESULTS AND CONCLUSION: (1) The mNss score of the treatment group was significantly lower than the control group at 12, 21, and 30 days, which indicated that radial extracorporeal shock wave therapy could significantly improve neurological function in rats with cerebral ischemia. (2) Compared with the control group, the expression of Nestin and NSE in ischemic hippocampus of rats increased significantly in the treatment group, which indicated that radial extracorporeal shock wave therapy could promote the proliferation and differentiation of neural stem cells in hippocampus after cerebral ischemia. (3) Compared with the control group, the expression of Notch1 and Hes1 in the ischemic hippocampus of rats in the treatment group was higher than that in the control group, which suggested that radial extracorporeal shock wave therapy could upregulate the expression of Notch1/Hes1 signaling pathway in the hippocampus. (4) The results confirm that radial extracorporeal shock wave therapy can promote the proliferation and differentiation of neural stem cells in the hippocampus after cerebral ischemia, and improve neurological function. The precise mechanism is related to the upregulation of Notch1/Hes1 signaling pathway. 


Key words: stem cells, neural stem cells, radial extracorporeal shock wave therapy, Notch1, signaling pathway, hippocampus, rat

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