CAJ | 학술논문

目的探讨嘌呤霉素敏感的氨肽酶(PSA)对β-淀粉样蛋白(Aβ25-35)诱导的SH-SY5Y细胞、PC12细胞凋亡的影响. 方法采用MTT法检测Aβ25-35对SH-SY5Y、PC12细胞生长的影响；Hoechst染色检测Aβ25-35对SH-SY5Y、PC12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PC12细胞中瞬时转染PSA重组质粒,加入Aβ25-35作用24h后收集细胞,进行流式细胞术检测细胞凋亡率；Western blotting检测PSA、caspase-3蛋白的表达变化；酶标仪检测caspase-3活性. 结果 Aβ25-35抑制SH-SY5Y、PC12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Aβ25-35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,PC12细胞中过表达的PSA能使Aβ25-35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 Aβ25-35能抑制神经细胞生长,引起神经细胞凋亡；PSA能抑制Aβ25-35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.
목적 탐토표령매소민감적안태매(PSA)대β-정분양단백(Aβ25-35)유도적SH-SY5Y세포、PC12세포조망적영향. 방법 채용MTT법검측Aβ25-35대SH-SY5Y、PC12세포생장적영향；Hoechst염색검측Aβ25-35대SH-SY5Y、PC12세포핵형태적영향；진일보채용지질체전염법재SH-SY5Y세포중순시전염PSA-siRNA,재PC12세포중순시전염PSA중조질립,가입Aβ25-35작용24h후수집세포,진행류식세포술검측세포조망솔；Western blotting검측PSA、caspase-3단백적표체변화；매표의검측caspase-3활성. 결과 Aβ25-35억제SH-SY5Y、PC12세포증식,세포핵발생조망적형태학개변,류식검측세포조망솔증가；재SH-SY5Y세포중,침묵PSA표체능사Aβ25-35유도적세포조망솔승고,촉진caspase-3매원격활,caspase-3활성승고.반지,PC12세포중과표체적PSA능사Aβ25-35유도적세포조망솔하강,억제caspase-3매원격활,caspase-3활성강저. 결론 Aβ25-35능억제신경세포생장,인기신경세포조망；PSA능억제Aβ25-35유도적신경세포조망,대신경원구유보호작용,기궤제가능여억제caspase-3통로적격활유관.
Objective To explore the effect of puromycin-sensitive aminopeptidase (PSA) on apoptosis induced by beta-amyloid peptides 25-35 (Aβ25-35) in SH-SY5Y cells and PC12 cells.Methods The effect of Aβ25-35 on proliferation of SH-SY5Y cells and PC12 cells was measured using MTT assay.Morphological changes of the cell nuclei induced by Aβ25-35 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 Aβ25-35.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 Aβ25-35 inhibited the growth of SH-SY5Y cells and PC 12 cells.Morphological changes indicated that apoptosis happened after cells were treated with Aβ25-35 for 24 h,as shown by flow cytometer.The apoptotic rate induced by Aβ25-35 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 Aβ25-35 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 Aβ25-35 could inhibit growth and induce apoptosis in SH-SY5Y cells and PC12 cells.PSA has protective effect on nerve cells by inhibiting apoptosis induced by Aβ25-35,and this effect may be ascribed to the inhibition ofpro-caspase-3 activation.