中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (3): 452-461.doi: 10.4103/1673-5374.245469

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

局灶性脑缺血再灌注：造模前3h Notch信号通路抑制剂DAPT预处理的神经保护

  

  • 出版日期:2019-03-15 发布日期:2019-03-15
  • 基金资助:

    国家自然科学基金(81660243); 中国贵州省科技厅社会发展科技计划项目(SY [2015] 3041); 中国贵州省科技厅项目（LG [2012] 028）

Neuroprotective effect of Notch pathway inhibitor DAPT against focal cerebral ischemia/reperfusion 3 hours before model establishment

Jun-Jie Wang, Jun-De Zhu , Xian-Hu Zhang, Ting-Ting Long, Guo Ge, Yan Yu   

  1. Department of Anatomy, School of Basic Medicine, Guizhou Medical University, Guian New District, Guizhou Province, China
  • Online:2019-03-15 Published:2019-03-15
  • Contact: Jun-De Zhu, MS, jdzhu73@126.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81660243; a grant from the Social Development Science and Technology Plan Project of Science and Technology Department of Guizhou Province of China, No. SY [2015] 3041; a grant from the Science and Technology Department of Guizhou Province of China, No. LG [2012] 028.

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摘要：

研究显示，Notch信号通路抑制剂N-[N-（3,5-二氟苯乙酰基）-1-丙氨酰] -S-苯基甘氨酸叔丁酯（DAPT）可减轻脑组织缺血性损伤。为探讨脑缺血再灌注损伤后DAPT的神经保护作用，实验在阻塞小鼠右侧大脑中动脉建立局灶性脑缺血再灌注模型前3h腹腔注射DAPT溶液。以Longa评估小鼠神经功能；以Nissl染色和TUNEL染色检测右前额叶皮质神经元损伤和细胞凋亡，以免疫荧光染色用于检测右前额叶皮质中的胶质纤维酸性蛋白和Notch1阳性细胞，以Western blot法检测检测右前额叶皮质中Hes1和Hes5蛋白的表达。见DAPT可明显改善局灶性脑缺血再灌注小鼠行为，并减轻神经元形态损伤，DAPT可降低右前额叶皮质胶质纤维酸性蛋白和Notch1阳性细胞数，减少凋亡细胞数量，降低白细胞介素6和肿瘤坏死因子α含量，同时下调Hes1和Hes5蛋白表达。结果说明DAPT可减轻脑缺血再灌注损伤后的脑组织损害，增强抗炎能力，有潜力成为预防性减轻脑缺血再灌注损伤的药物。

orcid: 0000-0001-9666-8010(Jun-De Zhu)

关键词: DAPT, 脑缺血, 胶质纤维酸性蛋白, Notch1, HES1, HES5

Abstract:

As an inhibitor of the Notch signaling pathway, N-[N-(3,5-difluorohenacetyl)-l-alanyl]-S-phenylglycine tert-butyl ester (DAPT) may protect brain tissue from serious ischemic injury. This study aimed to explore neuroprotection by DAPT after cerebral ischemia/reperfusion (I/R) injury. DAPT was intraperitoneally injected 3 hours before the establishment of a focal cerebral I/R model in the right middle cerebral artery of obstructed mice. Longa scores were used to assess neurological changes of mice. Nissl staining and TdT-mediated dUTP-biotin nick-end labeling staining were used to examine neuronal damage and cell apoptosis in the right prefrontal cortex, while immunofluorescence staining was used to detect glial fibrillary acidic protein- and Notch1-positive cells. Protein expression levels of Hes1 and Hes5 were detected by western blot assay in the right prefrontal cortex. Our results demonstrated that DAPT significantly improved neurobehavioral scores and relieved neuronal morphological damage. DAPT decreased the number of glial fibrillary acidic protein- and Notch1-positive cells in the right prefrontal cortex, while also reducing the number of apoptotic cells and decreasing interleukin-6 and tumor necrosis factor-α contents, and simultaneously downregulating Hes1 and Hes5 protein expression. These findings verify that DAPT alleviates pathological lesions and strengthens the anti-inflammatory response after cerebral I/R injury. Thus, DAPT might be developed as an effective drug for the prevention of cerebral I/R injury.

Key words: nerve regeneration, DAPT, cerebral ischemia, glial fibrillary acidic protein, Notch1, Hes1, Hes5, neural regeneration