嘌呤霉素敏感的氨肽酶对β-淀粉样蛋白诱导的细胞凋亡的影响  

Influence of puromycin-sensitive aminopeptidase in cell apoptosis induced by beta-amyloid peptides

作　　者：刁雪芹[1] 王苗苗[1] 庄云龙[2] 田克立[1] 徐霞[1] 褚倩倩[1] 周楠[1] 任桂杰[1] 

机构地区：[1]山东大学医学院生物化学与分子生物学研究所,济南250012 [2]山东省血液中心,济南250014

出　　处：《中华神经医学杂志》2013年第4期347-353,共7页Chinese Journal of Neuromedicine

摘　　要：目的探讨嘌呤霉素敏感的氨肽酶（PSA）对p一淀粉样蛋白（A1325-35）诱导的SH—SY5Y细胞、PCI2细胞凋亡的影响。方法采用MTT法检测AB。3，对SH．SY5Y、PCI2细胞生长的影响；Hoechst染色检测A对SH．SY5Y、PCI2细胞核形态的影响；进一步采用脂质体转染法在SH．SY5Y细胞中瞬时转染PSA—siRNA，在PCI2细胞中瞬时转染PSA重组质粒，加入A125-35作用24h后收集细胞．进行流式细胞术检测细胞凋亡率；Westemblotting检测PSA、caspase一3蛋白的表达变化；酶标仪检测caspase．3活性。结果A[325。，抑制SH．SY5Y、PCI2细胞增殖，细胞核发生凋亡的形态学改变．流式检测细胞凋亡率增加；在SH．SY5Y细胞中，沉默PSA表达能使A132，诱导的细胞凋亡率升高，促进caspase-3酶原激活，caspase-3活性升高。反之，PCI2细胞中过表达的PSA能使A132；诱导的细胞凋亡率下降，抑制caspase-3酶原激活，caspase-3活性降低。结论A。能抑制神经细胞生长，引起神经细胞凋亡；PSA能抑制A，诱导的神经细胞凋亡，对神经元具有保护作用，其机制可能与抑制caspase-3通路的激活有关。Objective To explore the effect of pttromycin-sensitive aminopeptidase （PSA） on apoptosis induced by beta-amyloid peptides 25-35 （AI325.35） in SH-SY5Y cells and PC12 cells. Methods The effect of AI3~35 on proliferation of SH-SY5Y cells and PC12 cells was measured using MTT assay. Morphological changes of the cell nuclei induced by A^25_35 were observed after staining with Hoechst 33342. PSA-siRNA was transfected into SH-SY5Y cells and recombinant PSA plasmid was transfected into PC12 cells using liposome method. The transfected cells were collected at 24 h after being treated with A^25-35. The percentage of apoptosis was determined by flow cytometer. The expressions of PSA and caspase-3 were detected by Western blotting, and caspase-3 activity was measured by microplate reader. Results AI325-35 inhibited the growth of SH-SY5Y cells and PC12 cells. Morphological changes indicated that apoptosis happened after cells were treated with A1325_35 for 24 h, as shown by flow cytometer. The apoptotic rate induced by A1~35 was increased by PSA-siRNA in SH-SY5Y cells, the activation of pro-caspase-3 was promoted, and the activity of caspase-3 was enhanced. The apoptotic rate induced by AI3^-35 was declined by over expression of PSA in PC12 cells, the activation of pro-caspase-3 was inhibited, and the activity of caspase-3 was decreased. Conclusion AI325.35 could inhibit growth and induce apoptosis in SH-SY5Y cells and PC 12 cells. PSA has protective effect on nerve cells by inhibiting apoptosis induced by A125-35, and this effect may be ascribed to the inhibition ofpro-caspase-3 activation.

关 键 词：嘌呤霉素敏感的氨肽酶 神经细胞 B-淀粉样蛋白 细胞凋亡 天冬氨酸 半胱氨酸蛋白酶-3 

分 类 号：R749.16[医药卫生—神经病学与精神病学]