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

doi:10.3969/j.issn.0529-1356.2013.02.006

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

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* 

作者信息 +

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

WANG Qiong-sa¹ QI Shuang-shuang² ZHOU Peng³ZHANG Chi-hao¹ CUI Huai-rui³ CHEN Shi-xin³ SUN Chen-you ^3* 

Author information +

文章历史 +

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。

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.

导出引用

王琼仨戚双双周鹏张驰豪崔怀瑞陈世新孙臣友*. 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[J]. 解剖学报. 2013, 44(2): 176-181 https://doi.org/10.3969/j.issn.0529-1356.2013.02.006

WANG Qiong-sa, QI Shuang-shuang, ZHOU Peng, ZHANG Chi-hao, CUI Huai-rui, CHEN Shi-xin, SUN Chen-you*. Effect of allopregnanolone on the dopaminergic neurons in the substantia nigra of Alzheimer’s disease mice [J]. Acta Anatomica Sinica. 2013, 44(2): 176-181 https://doi.org/10.3969/j.issn.0529-1356.2013.02.006

参考文献

[1] Scarmeas N, Hadjigeorgiou GM, Papadimitriou A, et al. Motor signs during the course of Alzheimer disease ［J］. Neurology, 2004, 63 (6): 975-982. 

[2] Oddo S, Caccamo A, Shepherd JD, et al. Triple-transgenic model of Alzheimer’s disease with plaques and tangles: intracellular abeta and synaptic dysfunction ［J］. Neuron, 2003, 39 (3): 409- 421. 

[3] Burns JM, Galvin JE, Roe CM, et al. The pathology of the substantia nigra in Alzheimer disease with extrapyramidal signs ［J］. Neurology, 2005, 64 (8): 1397-1403. 

[4] Brinton RD, Wang JM. Therapeutic potential of neurogenesis for prevention and recovery from Alzheimer’s disease: allopregnanolone as a proof of concept neurogenic agent ［J］. Curr Alzheimer Res, 2006, 3 (3): 185-190. 

［5］ Kawai H, Kotake Y, Ohta S, et al. Dopamine transporter and catechol-O-methyltransferase activities are required for the toxicity of

1-(3’,4’-dihydroxybenzyl)-1,2,3, 4-tetrahydroisoquinoline ［J］. Chem Res Toxicol, 2000, 13 (12): 1294-1301.

［6］ Wu LSh, Wang L, Ma YQ, et al. The innervations of rat pineal body: study by using immunohistochemical and morphometric methods ［J］. Acta Anatomica Sinica, 2005, 36 (3): 288-291. (in Chinese)

吴亮生，王蕾，马玉琼，等. 大鼠松果体的神经支配—免疫组织化学及光镜体视学研究［J］. 解剖学报，2005, 36 (3): 288-291. 

［7］ Klunk WE, Engler H, Nordberg A, et al. Imaging brain amyloid in Alzheimer’s disease with the Pittsburgh compound-B ［J］. Ann Neurol, 2004, 55 (3): 306-319. 

［8］ Jimenez-Escrig A, Rabano A, Guerrero C, et al. New V272A presenilin 1 mutation with very early onset subcortical dementia and parkinsonism ［J］. Eur J Neurol, 2004, 11 (10): 663-669. 

［9］ Perez SE, Lazarov O, Koprich JB, et al. Nigrostriatal dysfunction in familial Alzheimer’s disease-linked APPswe/PS1DeltaE9 transgenic mice ［J］. J Neurosci, 2005, 25 (44): 10220-10229. 

［10］ O’ Neil JN, Mouton PR, Tizabi Y, et al. Catecholaminergic neuronal loss in locus coeruleus of aged female dtg APP/PS1 mice ［J］. J Chem Neuroanat, 2007, 34 (3-4): 102-107. 

［11］ German DC, Nelson O, Liang F, et al. The PDAPP mouse model of Alzheimer’s disease: locus coeruleus neuronal shrinkage ［J］. J Comp Neurol, 2005, 492 (4): 469- 476.［12］ Arias-Carrión O, Yamada E, Freundlieb N, et al. Neurogenesis in substantia nigra of parkinsonian brains ［J］? J Neural Transm Suppl, 2009, (73): 279-285. 

［13］ Clinton LK, Blurton-Jones M, Myczek K, et al. Synergistic interactions between abeta, tau, and alpha-synuclein: acceleration of neuropathology and cognitive decline ［J］. J Neurosci, 2010, 30 (21): 7281-7289. 

［14］ Wang JM, Johnston PB, Ball BG, et al. The neurosteroid allopregnanolone promotes proliferation of rodent and human neural progenitor cells and regulates cell cycle gene and protein expression ［J］. J Neurosci, 2005, 25 (19): 4706-4718.

［15］ Wang JM, Sun C. Calcium and neurogenesis in Alzheimer’s disease ［J］. Front Neurosci, 2010, 4: 194. 

［16］ Brinton RD, Wang JM. Preclinical analyses of the therapeutic potential of allopregnanolone to promote neurogenesis in vitro and in vivo in transgenic mouse model of Alzheimer’s disease ［J］. Curr Alzheimer Res, 2006, 3 (1): 11-17. 

［17］ Bjugstad KB, Teng YD, Redmond DE Jr, et al. Human neural stem cells migrate along the nigrostriatal pathway in a primate model of Parkinson’s disease ［J］. Exp Neurol, 2008, 211 (2): 362-369. 