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目的观察热休克蛋白27(Hsp27)在蛋白酶体抑制剂PSI诱导PC12细胞帕金森病(PD)模型早期的表达变化,为深入研究PD的发病机制提供理论依据。方法在PC12细胞中加入终浓度为10μmol/L的PSI,建立PD细胞模型,通过吖啶橙(AO)和溴化乙啶(EB)及HE染色进行鉴定;PSI作用3h后提取蛋白,应用荧光差异凝胶电泳(DIGE)系统获得差异蛋白点,运用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOFProMS)鉴定差异蛋白。结果 PSI作用PC12细胞24h后,细胞内嗜酸性类Lewy小体形成,并发生细胞凋亡。PSI作用3h后,有6个蛋白点表达量显著变化,其中4个蛋白点表达量增加,2个蛋白点表达量降低。蛋白点1经质谱鉴定为Hsp27,表达量增加了1.74倍。结论在泛素-蛋白酶体系统(UPS)功能障碍引起PD细胞模型的早期病理变化过程中,可能通过诱导Hsp27表达量明显增加,从而抑制蛋白酶体抑制剂所引起的细胞毒性。
Objective To observe the expression changes of heat shock protein 27 (Hsp27) in the early stage of Parkinson’s disease (PD) induced by proteasome inhibitor PSI in PC12 cells and to provide a theoretical basis for further study on the pathogenesis of PD. Methods PC12 cells were treated with PSI at a final concentration of 10 μmol / L to establish a PD cell model and identified by acridine orange (AO), ethidium bromide (EB) and HE staining. After PSI for 3 hours, Differential protein spots were obtained by differential gel electrophoresis (DIGE) system, and differential proteins were identified by using MALDI-TOF-MS (matrix-assisted laser desorption / ionization time-of-flight mass spectrometry). Results After treated with PSI for 24h, PC12 cells formed eosinophilic Lewy bodies and apoptosis occurred. After PSI for 3h, the expression of 6 protein spots changed significantly, of which 4 protein spots increased and 2 spots decreased. Protein spot 1 was identified as Hsp27 by mass spectrometry, an increase of 1.74 fold. Conclusion During the early pathological changes of PD cell model induced by dysfunction of ubiquitin-proteasome system (UPS), the expression of Hsp27 may be significantly increased and the cytotoxicity induced by proteasome inhibitor may be inhibited.