CAJ | 학술논문

目的：探讨染料木素(genistein,GEN)通过激活 JNK调控Fas通路抑制Aβ(25-35)诱导的PC12细胞损伤凋亡的作用及分子机制。方法建立Aβ(25-35)诱导的PC12细胞模型,MTT法和流式细胞仪法测定细胞活力和凋亡率,荧光定量 PCR 检测 Fas 凋亡通路相关基因 Fas、FasL、caspase-3和caspase-8 mRNA相对表达情况,分光光度法检测caspase-3和caspase-8酶活性,Western blot检测JNK和p-JNK蛋白表达水平变化。结果 GEN下调Aβ(25-35)诱导引起的Fas、FasL、caspase-3和caspase-8 mRNA水平的增加,抑制Aβ(25-35)诱导的caspase-3和caspase-8酶活性,且明显降低Aβ(25-35)诱导的JNK磷酸化水平。结论GEN通过降低Aβ(25-35)诱导的 JNK磷酸化激活,调控JNK依赖的 Fas 凋亡通路,从而抑制 Aβ(25-35)诱导的PC12细胞凋亡,发挥神经保护作用。
목적：탐토염료목소(genistein,GEN)통과격활 JNK조공Fas통로억제Aβ(25-35)유도적PC12세포손상조망적작용급분자궤제。방법건립Aβ(25-35)유도적PC12세포모형,MTT법화류식세포의법측정세포활력화조망솔,형광정량 PCR 검측 Fas 조망통로상관기인 Fas、FasL、caspase-3화caspase-8 mRNA상대표체정황,분광광도법검측caspase-3화caspase-8매활성,Western blot검측JNK화p-JNK단백표체수평변화。결과 GEN하조Aβ(25-35)유도인기적Fas、FasL、caspase-3화caspase-8 mRNA수평적증가,억제Aβ(25-35)유도적caspase-3화caspase-8매활성,차명현강저Aβ(25-35)유도적JNK린산화수평。결론GEN통과강저Aβ(25-35)유도적 JNK린산화격활,조공JNK의뢰적 Fas 조망통로,종이억제 Aβ(25-35)유도적PC12세포조망,발휘신경보호작용。
Aim To investigate the effect of genistein ( GEN) against Aβ( 25 -35 )-induced PC12 cells in regulation of Fas pathway through the activation of JNK. Methods Aβ( 25 -35 )-induced PC12 cells model was established. MTT and fluorescence activated cell sorting to analyze cell viability and apoptotic rate. Fluorescence quantitative PCR was used to detect Fas apoptotic pathways related gene Fas, FasL, caspase-3 and caspase-8 mRNA relative expression. Spectropho-tometry was used to detect caspase-3 and caspase-8 en-zyme activity. Western blot was adopted to detect JNK and p-JNK protein expression level changes. Results GEN attenuated Aβ( 25-35 )-induced upregulation of Fas and FasL, caspase-3 and caspase-8 mRNA lev-el, caspase-3 and caspase-8 enzyme activity, and sig-nificantly reduced Aβ(25-35) induced JNK phospho-rylation level. Conclusion GEN can protect PC12 cells from Aβ(25-35)-induced apoptosis via reducing Aβ( 25 -35 )-induced phosphorylation of JNK activa-tion, and then inhibit the JNK dependent Fas apoptotic pathway.