【摘要】 目的探讨蛋白酶体抑制剂诱导PC12细胞凋亡的作用通路,为深入研究泛素-蛋白酶体系统(UPS)功能障碍诱发细胞凋亡发病机制提供新线索。方法建立PSI诱导的PC12细胞模型,在48h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)鉴定出差异蛋白。结果实验组与对照组比较,在48h看到细胞凋亡(细胞核呈固缩状或碎裂状),凋亡率达14.86%。实验组钙激活蛋白酶(calpain)与对照组比较表达量显著增高,有统计学意义(P<0.05)。结论首次发现在蛋白酶体抑制PC12细胞模型中calpain蛋白表达显著增高。结合GRP78和GRP94表达上调的发现提示内质网应激(ERS)在UPS功能障碍引起细胞凋亡过程中扮演重要角色。更多还原

【Abstract】 Objective To explore the action pathway of proteasomes inhibitor (PSI) induced PC12 apoptosis to provide new clue for studying pathogenesis of ubiquitin-proteasomes system (UPS) dysfunction induced apoptosis. Methods The protein of PC12 cell model induced by PSI was extracted 48 h later to obtain difference protein site by fluorescence differential gel electrophoresis (DIGE) and to identify difference protein by MALDI-TOF Pro MS. Results In experimental group, cell apoptosis was seen, presenting pyknosis or chipped cellular nucleus, apoptosis rate was 14.86%. Calpain expressions in experimental group were significantly higher than that in control group(P<0.05).Conclusions The increased calpain expressions in PSI-induced PC12 cell model are found firstly, combining the upregulated GRP78 and GRP94 expressions could indicate that endocytoplasmic reticulum stress (ERS) play an important role in cell apoptosis induced by UPS dysfunction.更多还原