MPTP慢性帕金森病小鼠纹状体差异蛋白质的研究

目的分离、鉴定MPTP诱导慢性帕金森病模型小鼠纹状体差异表达的蛋白质,对MPTP慢性PD动物模型的特异性蛋白质组进行初步探讨,为PD的发病机制提供一定的蛋白质组学依据。方法成功建立MPTP诱导慢性帕金森病小鼠模型,提取模型组和对照组小鼠脑纹状体蛋白质,分别以固相pH梯度等电聚焦为第一向,SDS-PAGE垂直电泳为第二向进行2-DE。图像分析软件PDQUEST8.0分析电泳图谱找出差异表达蛋白,运用MALDI-TOF MS质谱鉴定;其肽质量指纹图（PMF）经MS Fit检索。结果比较MPTP诱导慢性PD模型小鼠和正常对照小鼠纹状体二向电泳图,发现12个蛋白表达异常,最终鉴定出其中4个蛋白质：线粒体裂殖调节因子1（mitochondrial fission regulator 1）、类泛素样蛋白3前体（ubiquitin-like protein 3 precursor）表达下调;S100蛋白A10（proteinS100-A10）、Lin-7 homolog B为新出现点。结论初步鉴定出MPTP慢性PD模型小鼠纹状体部分差异表达蛋白,所发现4个表达异常的蛋白质与帕金森病线粒体的损伤和兴奋性神经毒性密切相关,与PD的发病机制相符,为深入研究帕金森病病理机制奠定了基础。

Proteomic Analysis of Striatum in the MPTP-Induced Mouse Model of Parkinson's Disease

Objective To compare the striatal proteins between a mouse model of Parkinson＇s disease（PD） induced by 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP） administration and the control by proteomics technology,and to lay a foundation for understanding the disease-specific proteins in Parkinson＇s disease.Methods To establish a mouse model of Parkinsonism induced by low dose of MPTP injection.Using urea-based buffer to extract the striatal proteins and run through immobilized pH gradient（IPG） isoelectric focusing electrophoresis as the first dimension,and then run through vertical SDS-PAGE electrophoresis as the second dimension.Protein profiles in the tissue of control and treated mice were compared by the PDQuest 2D-gel analysis software（BIO-Rad laboratory）.The protein spots showing differential expression were analysed by matrix assisted laser desorption/ionizing time of flight（MALDI-TOF） mass spectrometry.Results In the striatum,12 proteins were differentially expressed in PD mouse compared with normal control mice.There were 4 spots identified definitely： mitochondrial fission regulator 1 and ubiquitin-like protein 3 precursor were significantly decreased,and protein S100-A10 and Lin-7 homolog B were only found in the PD mouse group.Conclusion Four proteins are differentially expressed in the striatum of MPTP-induced mouse models of Parkinson＇s disease by proteomic analysis. Our findings further support our previous studies describing the same proteins as a result of the damage of mitochondrial function and the excitatory neurotoxicity in PD.The present study laid a foundation for the further investigation of pathogenesis of Parkinson＇s disease.