解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (2 ): 176-181.doi: 10.3969/j.issn.0529-1356.2013.02.006

• 神经生物学 • 上一篇    下一篇

别异烯醇酮对阿尔茨海默病小鼠黑质多巴胺神经元的影响

王琼仨1  戚双双2  周鹏3  张驰豪1  崔怀瑞3  陈世新3  孙臣友3*    

  1. 1.温州医学院第一临床医学院，浙江 温州 325035; 2. 温州医学院附属第二医院药剂科，浙江 温州 325000; 3.温州医学院人体解剖学教研室，浙江 温州 325035
  • 收稿日期:2012-07-05 修回日期:2012-08-24 出版日期:2013-04-06 发布日期:2013-04-06
  • 通讯作者: 孙臣友 E-mail:sunchenyou1972@yahoo.com.cn
  • 基金资助:

    省自然科学基金资助项目;自然科学基金资助项目

Effect of allopregnanolone on the dopaminergic neurons in the substantia nigra of Alzheimer’s disease mice 

WANG Qiong-sa1 QI Shuang-shuang2 ZHOU Peng3 ZHANG Chi-hao1 CUI Huai-rui3 CHEN Shi-xin3 SUN Chen-you 3*    

  1. 1. the First Clinical Medical College, Wenzhou Medical College, Zhejiang Wenzhou 325035, China; 2. Department of Pharmacy, Second Affiliated Hospital of Wenzhou Medical College, Zhejaing Wenzhou 325000, China; 3. Department of Anatomy, Wenzhou Medical College, Zhejiang Wenzhou 325035, China
  • Received:2012-07-05 Revised:2012-08-24 Online:2013-04-06 Published:2013-04-06

摘要：

目的 通过分析黑质致密部（SNpc）多巴胺神经元和新生的多巴胺神经元的数目，评估别异烯醇酮（APα）对阿尔茨海默病（AD）小鼠多巴胺神经元的增生是否有影响。方法 选用3月龄雄性淀粉样前体蛋白/早老蛋白（APP/PS1）双转基因AD小鼠（2xTg-AD）作为实验组，背景非转基因小鼠（Non-Tg）作为对照组。2xTg-AD小鼠与Non-Tg小鼠分别行APα和PBS处理，24d后，每组6只小鼠做行为学检测；每组另6只小鼠取出脑组织后固定，通过免疫组织化学、体视测量学和吸光度测定，观察和分析各组小鼠SNpc标记酪氨酸羟化酶（TH）的多巴胺神经元的数目及其表达情况；并通过免疫荧光染色技术计算各组小鼠双标记的TH/5-溴脱氧尿嘧啶核苷（BrdU）阳性神经元的数目。结果 与Non-Tg溶剂对照组小鼠相比，2xTg-AD溶剂对照组小鼠开野实验移动格子的数目明显减少（P ＜0.01）；SNpc区域TH的免疫活性强度明显降低（P ＜0.05）；多巴胺神经元的数目明显减少（ P ＜0.05）；BrdU/TH双标记神经元的数目均明显降低（ P ＜0.05）。与2xTg-AD溶剂对照组小鼠相比，经APα处理的2xTg-AD小鼠移动格子的数目明显增加（ P ＜0.05）；SNpc TH阳性神经元的数目明显增加（ P ＜0.05）；BrdU/TH双标记的阳性神经元的数目亦明显增多（P ＜0.01）。但是，TH免疫活性强度无显著性变化（ P ＞0.05）。结论 APα通过增加AD小鼠SNpc新生的多巴胺的数目达到改善其多巴胺神经元减少和行为学下降的作用，表明APα是一种潜在性的神经增生的促进剂。

关键词: 别异烯醇酮 , 多巴胺神经元 , 神经增生 , 阿尔茨海默病 , 免疫组织化学 , 免疫荧光染色 ,  淀粉样前体蛋白/早老蛋白双转基因AD小鼠

Abstract:

Objective The present study was undertaken to evaluate the effects of allopregnanolone (APα) on neurogenesis of dopaminergic neurons by analyzing the numbers of dopaminergic neurons and newly formed dopaminergic neurons in the substantia nigra pars compacta (SNpc) of Alzheimer’s disease (AD) mice. Methods Experiments were performed using 3-month-old male double transgenic amyloid precursor protein with presenilin (APP/PS1) mouse model of AD (2xTg-AD) and non-transgenic (non-Tg) background mice. These mice were administered with APα or phosphate buffer saline (PBS), respectively. Following 24 days, six mice in each group were performed behavior testing by open-field test. The brain tissue from another 6 mice in each group was taken out and post-fixed and the numbers of tyrosine hydroxylase (TH) expressing dopaminergic neurons and TH expression were observed and analyzed in the SNpc region by the immunohistochemistry, unbiased stereology and optical intensity. In addition, the numbers of TH/bromodeoxyuridine (BrdU) double labeled dopaminergic neurons were counted by the immunofluorescent method. Results As compared with the Non-Tg+PBS mice, the data for 2xTg-AD+PBS mice were obviously decreased in the ambulation of open-field test (P ＜0.01), the intensities of TH immunohistochemical activity (P ＜0.05), the numbers of dopaminergic neurons and BrdU/TH double labeled neurons in the SNpc regions (P ＜0.05). As compared with the 2xTg-AD+PBS mice, the data for 2xTg-AD+APα mice was obviously increased in the ambulation of openfield test (P ＜0.05), the numbers of dopaminergic neurons ( P ＜0.05) and BrdU/TH double labeled neurons in the SNpc regions ( P ＜0.01). However, the intensities of TH immunohistochemical activity didn’t alter significantly between the 2xTg-AD+PBS and 2xTg-AD+APα mice (P＞0.05). Conclusion These results indicate that APα may ameliorate the loss of numbers of dopaminergic neurons by increasing the numbers of newly formed dopaminergic neurons in the SNpc of 2xTg-AD mice. These results suggest the APα may have a potential therapeutic efficacy in neurogenesis.

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