Notch信号通路对脑缺血大鼠神经干细胞移植后神经再生的影响

doi:10.16098/j.issn.0529-1356.2020.01.003

解剖学报 ›› 2020, Vol. 51 ›› Issue (1): 15-20.doi: 10.16098/j.issn.0529-1356.2020.01.003

Notch信号通路对脑缺血大鼠神经干细胞移植后神经再生的影响

刘宗秀^1,2 张紫微² 成媛² 靳晓飞² 周晓红² 高维娟^2*

1. 承德医学院病理生理学教研室，河北承德 067000; 2.河北中医学院河北省心脑血管病中医药防治重点实验室，石家庄 050091

收稿日期:2019-01-09 修回日期:2019-02-16 出版日期:2020-02-06 发布日期:2020-04-21
通讯作者: 高维娟 E-mail:gwj6088@163.com
基金资助:
河北省政府资助临床医学优秀人才培养和基础课题研究项目;河北省教育厅青年基金;河北省中医药管理局课题

Effect of Notch signaling pathway on nerve regeneration after cerebral ischemia in rats with neural stem cells transplantation#br#

LIU Zong-xiu^1,2 ZHANG Zi-wei² CHENG Yuan² JIN Xiao-fei²ZHOU Xiao-hong² GAO Wei-juan^2*

1. Department of Pathophysiology, Chengde Medical College, Hebei Chengde 067000, China; 2. Hebei Key Laboratory of Chinese Medicine Basic Research on Prevention and Treatment of Cardio-cerebrovascular Disease, Hebei University of Chinese Medicine, Shijiazhuang 050091, China

Received:2019-01-09 Revised:2019-02-16 Online:2020-02-06 Published:2020-04-21
Contact: GAO Wei-juan E-mail:gwj6088@163.com

摘要/Abstract

摘要：

目的探讨抑制Notch信号通路促进脑缺血大鼠神经干细胞（NSCs）移植后神经再生的机制。方法采用大脑中动脉阻塞MCAO）方法构建局灶性脑缺血模型，体外培养NSCs，移植入纹状体缺血区。将40只SD大鼠随机分为假手术组、模型组、移植组（移植神经干细胞）、氮-［氮-(3,5-二氟苯乙酰)-L-丙氨酰］-S-苯基甘氨酸丁酯］（DAPT）+移植组。采用HE染色观察各组大鼠神经元损伤程度，利用免疫组织化学、Western blotting检测各组大鼠脑组织中Notch1、Hes1及Hes5的表达情况。结果与假手术组相比，模型组神经元损伤严重，出现核固缩和核溶解，Notch1、Hes1及Hes5阳性细胞表达明显增多，Notch1、Hes1及Hes5蛋白表达显著上调（P＜0.05）。与模型组相比，移植组和DAPT+移植组神经元损伤均不同程度缓解，Notch1、Hes1及Hes5阳性细胞均有部分表达，各项蛋白表达均有所降低（P＜0.05）；DAPT+移植组神经元损伤明显恢复，较移植组各项蛋白阳性细胞和蛋白表达进一步降低(P＜0.05)。结论抑制Notch信号通路可促进脑缺血大鼠神经干细胞移植后的神经再生，其机制主要与下调Notch1、Hes1及Hes5的表达有关。

关键词: 脑缺血, 神经干细胞移植, 神经再生, Notch信号通路, HE染色, 大鼠

Abstract:

Objective To investigate the mechanism of inhibiting the Notch signaling pathway in promoting neural regeneration after neural stem cells (NSCs) transplantation in rats with cerebral ischemia. Methods The model of focal cerebral ischemia was established by middle cerebral artery occlusion (MCAO) method . NSCs were cultured in vitro and transplanted into the striatum ischemic area. In the experiment, 40 SD rats were divided into sham operation group, model group, transplantation group (transplanted neural stem cells), and N-［N-(3,5-difluorohenacetyl)-L-alanyl］-S-phenylglycinet-butyl ester（DAPT）+ transplantation group. The degree of neuronal damage in each group was observed by HE staining. The expressions of Notch1, Hes1 and Hes5 in the brain tissue of each group were detected by immunohistochemistry and Western blotting. Results Compared with the sham operation group, the neuron injury in the model group was severe, and nuclear pyknosis and nuclear lysis were observed. The Notch1, Hes1 and Hes5 positive cells increased significantly, and the expression of Notch1, Hes1 and Hes5 proteins were significantly up-regulated (P<0.05). Compared with the model group, the neuronal damage in the transplantation group and the DAPT+ transplantation group were all relieved, and the Notch1, Hes1 and Hes5 positive cells were partially expressed, and the expression of each protein decreased (P<0.05). The DAPT+ transplantation group neurons were compared. The damage was obviously restored, and the expression of protein-positive cells and protein further decreased (P<0.05). Conclusion Inhibition of Notch signaling pathway can promote nerve regeneration after neural stem cell transplantation in rats with cerebral ischemia. The mechanism is mainly related to down-regulation of Notch1, Hes1 and Hes5 expression.

Key words: Cerebral ischemia, Neural stem cell transplantation, Nerve regeneration, Notch signaling pathway, HE staining, Rat

中图分类号:

R743
R363

刘宗秀张紫微成媛靳晓飞周晓红高维娟. Notch信号通路对脑缺血大鼠神经干细胞移植后神经再生的影响[J]. 解剖学报, 2020, 51(1): 15-20.

LIU Zong-xiu ZHANG Zi-wei CHENG Yuan JIN Xiao-fei ZHOU Xiao-hong GAO Wei-juan. Effect of Notch signaling pathway on nerve regeneration after cerebral ischemia in rats with neural stem cells transplantation#br#[J]. Acta Anatomica Sinica, 2020, 51(1): 15-20.

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Notch信号通路对脑缺血大鼠神经干细胞移植后神经再生的影响

Effect of Notch signaling pathway on nerve regeneration after cerebral ischemia in rats with neural stem cells transplantation#br#

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