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目的通过比较鱼藤酮和1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)两种慢性帕金森病(Parkinson’s disease,PD)模型小鼠与对照组小鼠黑质蛋白表达谱,找出两种PD模型小鼠共同的差异蛋白,探索筛选出PD的疾病特异性蛋白。方法构建鱼藤酮和MPTP慢性PD小鼠模型组以及其各自的对照组1和对照组2,并进行行为学评价以及黑质酪氨酸羟化酶(tyrosine hydroxylase,TH)免疫组化。分别挖取黑质,提取总蛋白,进行双向凝胶电泳。运用PDQuest 8.0图像分析软件对鱼藤酮组与对照组1、MPTP与对照组2的电泳图谱分别进行分析,找出各自的差异表达蛋白。差异蛋白酶切后,MALDI-TOF-MS进行质谱鉴定。最后将所得到的肽质量指纹图(PMF)进行数据库搜索以及生物信息学分析。结果鱼藤酮组与对照组1、MPTP组与对照组2小鼠间行为学差异具有统计学意义(P<0.05),且黑质TH阳性细胞计数差异也具有统计学意义(P<0.05)。鱼藤酮组与对照组1比较发现有22个差异蛋白,MPTP组与对照组2比较发现有28个差异蛋白,其中有7个为共同的差异蛋白。我们成功鉴定其中3个共同的差异蛋白:PPP2R1A、吡哆醛激酶、peroxiredoxin-2。结论鉴定出3个在PD蛋白质组学中少见报道的蛋白。这些蛋白水平的上调或下调可能与PD的发病密切相关,可作为PD的疾病特异性蛋白。
Objective To compare the effects of rotenone and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) Parkinson’s disease (PD) model mice and control mice were used to find out the common proteins in the two PD mice and explore the disease-specific proteins of PD. Methods The rotenone and MPTP chronic PD mice model group and their control group 1 and control group 2 were constructed and their behavioral evaluation and immunohistochemistry of tyrosine hydroxylase (TH) were performed. Separately dig the substantia nigra and extract the total protein for two-dimensional gel electrophoresis. The PDQuest 8.0 image analysis software was used to analyze the electrophoresis patterns of rotenone group and control group 1, MPTP and control group 2, respectively, to find their respective differentially expressed proteins. After different protease digestion, MALDI-TOF-MS was identified by mass spectrometry. Finally, the obtained peptide mass fingerprint (PMF) database search and bioinformatics analysis. Results The rotenone group and the control group 1, MPTP group and control group 2 mice behavioral differences were statistically significant (P <0.05), and the substantia nigra TH positive cell count was also statistically significant difference (P <0.05). There were 22 differential proteins in rotenone group and control group 1, 28 differential proteins in MPTP group and control group 2, of which 7 were common differential proteins. We successfully identified three of these proteins in common: PPP2R1A, pyridoxal kinase, peroxiredoxin-2. Conclusion Three rare proteins were identified in PD proteomics. The up-regulation or down-regulation of these protein levels may be closely related to the pathogenesis of PD and may serve as disease-specific proteins of PD.