利用FRET技术在活细胞内研究红景天甙对Aβ_(25-35)诱导PC12细胞凋亡的抑制作用

INHIBITIONS OF SALIDROSIDE ON THE Aβ_(25-35)-INDUCED APOPTOSIS BY FRET IN LIVING PC12 CELLS

为研究红景天甙(salidroside)对-淀粉样肽25-35(.amyloidpeptide25-35,A/25-3)5诱导PC12细胞凋亡的抑制作用,采用CellCountingKit-8(CCK-8)分析细胞的存活率,通过光镜检测细胞形态并配以Hoechst染色检测细胞核固缩,利用荧光共振能量转移(fluorescenceresonanceenergytransfer,FRET)技术在单个活细胞中检测caspase-3和caspase-8活性的动态变化。结果表明,红景天甙可剂量依赖性抑制A025-35引起的细胞凋亡,提高细胞的存活率;红景天甙对caspase-3的活性有明显的抑制作用,而且A125-35诱导细胞凋亡不依赖于caspase-8的激活。这些结果提示抑制caspase-3的活性是红景天甙抑制A225-35诱导PC12细胞凋亡的机制之一。

To investigate the inhibitions of salidroside on the apoptosis induced by Aβ25-35 in PC12 cells, cell survival was analyzed with cell counting Kit-8 （CCK-8）, cell morphology was observed by optical microscope, nuclear condensation was shown by Hoechst staining, the dynamics of caspase-3 and caspase-8 activation in single living cell were monitored by fluorescence resonance energy transfer （FRET） respectively. The results showed that salidroside elevated the cell survival by inhibiting the Aβ25-35-induced apoptosis in a dose-dependent manner, and that salidroside greatly inhibited the caspase-3 activation. Furthermore, the apoptosis induced by Aβ25-35 was independent of the caspase-8 activation. In conclusion, these data indicate that salidroside can protect the PC12 cells from Aβ25-35-induced apoptosis by inhibiting the activation of caspase-3.