AAV介导的α-synuclein基因过表达致多巴胺能神经元损伤——一种制作轻度帕金森病大鼠模型的新方法  

Overexpression of α-Synuclein Gene Caused Dopaminergic Neuron Damage in Substantia Nigra of Rats

作　　者：鲁玲玲[1] 周爱霞[1] 杨慧[1] 

机构地区：[1]首都医科大学北京神经科学研究所北京市神经再生修复研究重点实验室教育部神经变性病学重点实验室,北京100069

出　　处：《中国生物工程杂志》2009年第1期1-6,共6页China Biotechnology

基　　金：国家“973”计划(2006CB500706);国家自然科学基金(30670655);北京市教育委员会科技发展计划(KM200610025002);教育部高等学校博士学科点专项科研基金(20060025004);北京市优秀人才培养专项经费(20081d0501800210)资助项目

摘　　要：目的:研究过表达α-synuclein基因是否导致大鼠黑质纹状体选择性损伤,为帕金森病(parkinson’s disease,PD)大鼠模型的制备提供一种新的方法。方法:用腺相关病毒(adeno-associated virus,AAV)做载体,将人野生型α突触核蛋白(α-synuclein,NACP)引入大鼠脑内,观察大鼠行为学的改变,通过免疫组织化学染色观察其对黑质多巴胺能神经元细胞的影响,高效液相色谱(HPLC)检测纹状体多巴胺(DA)的含量。结果:α-synuclein基因过表达后大鼠出现自发性活动减少、爬行活动减慢、暂时性躯干震颤、竖毛等类似PD初期的症状和体征;大鼠脑黑质TH阳性神经元细胞随时间的延长出现数目减少,并且纹状体DA含量也出现减少,并且出现α-synuclein的积聚。结论:上述结果表明α-synuclein基因的过表达引起黑质多巴胺能神经元细胞的死亡,对大鼠的运动行为有一定的影响,产生类似于PD早期的症状与体征,与化学毒素(如6-OHDA,MPTP)诱导的动物模型相比,此法制作的动物模型可模拟PD缓慢发展的进程,为研究PD的病程进展及发病机制提供一个理想的动物模型。synuclein in rat method to make Objectiive：The present study was designed to explore whether overexpression of human wild α- brain caused selective dopaminergic neuron loss in substantia nigra and aimed to find out a new a rat model of Parkinson＇ s disease （PD）. Methods：The human wild α-synuclein gene was induced into the rat brain by Adeno-Associated Virus （AAV） vector. The overexpression of α-synuclein was detected by reahime PCR. The behavior of rats were recorded every 4 weeks after the viral particle injection. TH immunohistochemistry were performed at 4, 8, 12 and 16 weeks post-injection as well as the dopamine （DA）, 3,4- dihydroxypheny- lacetic acid （DOPAC） of striatum were determined by high performance liquid chromatography coupled with electrochemical detection. Results： Reahime PCR results revealed a significant overexpression of α-synuclein in the injected hemisphere. By 8 weeks post injection, a significant loss of the dopaminergic neurons was observed. 34% of the dopaminergic neurons were lost after 12 weeks, and about 60% cells loss after 16 weeks. The DA and DOPAC levels in the striatum decreased about 15% 12 weeks after injecting viral particle carried α-synuclein gene and 30% decreased after 16 weeks. The AAV-α-synuclein- treated rats developed a type of motor impairment, i. e. , nigrostriatal damage. Conclusion：All the results showed head position bias, compatible with this magnitude of that overexpression of human wild α-synuclein caused selective dopaminergic neuron loss and mimic a symptom of human PD in rats. This may be a new methed to make rat PD model which can offer new opportunities for the study of pathogenetic mechanismsand exploration of new therapeutic targets of particular relevance to human PD.

关 键 词：帕金森病 Α-突触核蛋白 动物模型 

分 类 号：Q789[生物学—分子生物学]