中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (4): 724-730.doi: 10.4103/1673-5374.266917

• 原著:退行性病与再生 • 上一篇    下一篇

多巴胺耗竭下纹状体中星形胶质细胞的特征性反应

  

  • 出版日期:2020-04-15 发布日期:2020-05-29
  • 基金资助:

    国家自然科学基金项目（81471288），国家高技术研究发展计划（863计划）（2017YFA0104704）

Characteristic response of striatal astrocytes to dopamine depletion

Yao-Feng Zhu1, 2, Wei-Ping Wang3, Xue-Feng Zheng1, Zhi Chen1, Tao Chen1, Zi-Yun Huang1, Lin-Ju Jia1, Wan-Long Lei1   

  1. 1 Department of Anatomy, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong Province, China
    2 Institute of Medicine, College of Medicine, Jishou University, Jishou, Hunan Province, China
    3 Periodical Center, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, China
  • Online:2020-04-15 Published:2020-05-29
  • Contact: Wan-Long Lei, PhD,leiwl@mail.sysu.edu.cn.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 81471288 (to WLL); the National High Technology Research and Development Program of China (863 Program), No. 2017YFA0104704 (to WLL).

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摘要：

星形胶质细胞和星形胶质细胞相关蛋白在维持正常脑功能以及调节脑疾病和脑损伤的病理过程中起重要作用，但是星形胶质细胞在多巴胺耗竭的纹状体中的作用尚不清楚。因此实验在大鼠右侧内侧前脑束中注射10μL6-羟基多巴胺（2.5μg/μL）诱导建立帕金森病模型，以免疫组化染色检测纹状体中胶质纤维酸性蛋白、钙结合蛋白B和信号转录因子3的免疫阳性反应，以及胶质纤维酸性蛋白与钙结合蛋白B和信号转录因子3的共表达情况，以Western blot检测纹状体中胶质纤维酸性蛋白、钙结合蛋白B和信号转录因子3的蛋白表达情况。结果发现：①纹状体星形胶质细胞为胶质纤维酸性蛋白阳性反应，多巴胺剥夺明显致使胶质纤维酸性蛋白阳性细胞呈现激活和增生性变化，其形态学变化明显；②在正常纹状体中可见钙结合蛋白B和信号转录因子3阳性反应，且钙结合蛋白B阳性细胞在纹状体分布均匀，胞体圆形，分支稀少。信号转录因子3阳性细胞呈圆球状，分支稀疏，可见一些染色加深和体积大的阳性细胞。多巴胺剥夺明显诱导纹状体钙结合蛋白B和信号转录因子3反应增强；③纹状体星形胶质细胞均表达钙结合蛋白B和信号转录因子3蛋白，而且纹状体多巴胺剥夺明显导致胶质纤维酸性蛋白/钙结合蛋白B和胶质纤维酸性蛋白/信号转录因子3双标记细胞数量增加；④6-羟基多巴胺所致的纹状体多巴胺剥夺明显诱导胶质纤维酸性蛋白、钙结合蛋白B以及信号转录因子3蛋白发生高表达。提示星形胶质细胞的形态变化以及星形胶质细胞的相关蛋白表达与纹状体多巴胺耗竭的病理过程有关。动物实验于2014年9月22日经中国中山大学动物护理与使用委员会批准，批准号中山医医伦2014第23号。

orcid: 0000-0002-1344-5215 (Wan-Long Lei)

关键词: 多巴胺耗竭, 星形胶质细胞, 钙结合蛋白B, 信号转录因子3, 帕金森病, 6-羟基多巴胺, 纹状体, 多巴胺能神经元, 神经再生

Abstract: Astrocytes and astrocyte-related proteins play important roles in maintaining normal brain function, and also regulate pathological processes in brain diseases and injury. However, the role of astrocytes in the dopamine-depleted striatum remains unclear. A rat model of Parkinson’s disease was therefore established by injecting 10 μL 6-hydroxydopamine (2.5 μg/μL) into the right medial forebrain bundle. Immunohistochemical staining was used to detect the immunoreactivity of glial fibrillary acidic protein (GFAP), calcium-binding protein B (S100B), and signal transducer and activator of transcription 3 (STAT3) in the striatum, and to investigate the co-expression of GFAP with S100B and STAT3. Western blot assay was used to measure the protein expression of GFAP, S100B, and STAT3 in the striatum. Results demonstrated that striatal GFAP-immunoreactive cells had an astrocytic appearance under normal conditions, but that dopamine depletion induced a reactive phenotype with obvious morphological changes. The normal striatum also contained S100B and STAT3 expression. S100B-immunoreactive cells were uniform in the striatum, with round bodies and sparse, thin processes. STAT3-immunoreactive cells presented round cell bodies with sparse processes, or were darkly stained with a large cell body. Dopamine deprivation induced by 6-hydroxydopamine significantly enhanced the immunohistochemical positive reaction of S100B and STAT3. Normal striatal astrocytes expressed both S100B and STAT3. Striatal dopamine deprivation increased the number of GFAP/S100B and GFAP/STAT3 double-labeled cells, and increased the protein levels of GFAP, S100B, and STAT3. The present results suggest that morphological changes in astrocytes and changes in expression levels of astrocyte-related proteins are involved in the pathological process of striatal dopamine depletion. The study was approved by Animal Care and Use Committee of Sun Yat-sen University, China (Zhongshan Medical Ethics 2014 No. 23) on September 22, 2014.

Key words: 6-hydroxydopamine, astrocyte, dopamine depletion, dopaminergic neurons, Parkinson’s disease, S100B, STAT3, striatum