【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。更多还原

【Abstract】 Objectiive:The present study was designed to explore whether overexpression of human wild α-synuclein in rat brain caused selective dopaminergic neuron loss in substantia nigra and aimed to find out a new method to make 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 realtime 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:Realtime 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.,head position bias,compatible with this magnitude of nigrostriatal damage.Conclusion:All the results showed 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.更多还原