【摘要】 目的探讨线粒体乙醛脱氢酶前体氧化修饰水平变化在帕金森病(PD)发病机制中的可能作用。方法实验分为实验组(PD细胞模型组)和对照组。PC12细胞孵育24 h后,实验组加入终浓度10μM的蛋白酶体抑制剂PSI(以DMSO溶解),对照组加入DMSO,继续孵育24 h后用于研究。MTT法检测细胞存活率,HE染色观察细胞内包含体的形成。提取细胞总蛋白,一向等电聚焦电泳后通过2,4-二硝基苯肼(DNP)的衍生步骤将含有羰基的氧化蛋白质标记上。而后通过2D凝胶电泳、Western印迹技术与质谱鉴定相结合的方法比较实验组和对照组线粒体乙醛脱氢酶前体氧化修饰水平的变化。结果与对照组比较,PD细胞模型中线粒体乙醛脱氢酶前体氧化修饰水平显著升高。结论线粒体乙醛脱氢酶前体氧化修饰水平变化可能为PD发病机制的研究提供新的线索。更多还原

【Abstract】 Objective To explore the possible effect of the oxidative modificational level of ALDH precursor in the pathogenesis of PD.Methods The cultured PC12 cells were divided into control group and 10μM PSI-treated group.MTT assay were used to test cell viability;HE stains were used to show intracytoplasmic inclusions.The specifically oxidized protein spots were obtained by coupling 2D gel electrophoresis with Western blot techniques,subsequently identified by MALDI-TOF mass spectrometry(MS).Results The oxidative modificational level of ALDH precursor increased evidently in the cell model of PD,compared with the vehicle group.Conclusion The changed oxidative level of ALDH precursor may provided new clue for further explaining the pathogenesis of PD.更多还原