FLZ Inhibited γ-secretase Selectively and Decreased Aβ Mitochondrial Production in APP-SH-SY5Y Cells

Amyloid precursor protein (APP) metabolism is a key factor in the pathogenesis of Alzheimer's disease (AD). Amyloid-beta (Aβ) in mitochondria comes from APP mitochondrial metabolism or from the uptake Aβ from outside of mitochondria. It has been recently proposed that mitochondria are involved in the biochemical pathways through which Aβ causes neuronal dysfunction. The accumulated Aβ in mitochondria decreases the level of cytochrome c oxidase (COX IV) and attenuates the ATP production consequently. FLZ is a synthetic cyclic derivative of squamosamide from Annona glabra. In this study, the effect of FLZ on APP processing in mitochondria was investigated in SH-SY5Y cells over-expressing APP695 (wt/Swe). FLZ treatment attenuated APP processing and decreased Aβ production in mitochondria. The mitochondrial function was increased with the upregulation of COX IV both at protein and activity levels. ATP production was also increased after FLZ treatment. The mechanistic study showed that FLZ inhibited γ-secretase activity by decreasing C-terminal fragment protein level of presenilin, the active center of γ-secretase. The effect of FLZ differs from DAPT (a non-selective γ-secretase inhibitor), suggesting FLZ is a selective γ-secretase inhibitor. FLZ selectively inhibited γ-secretase in the cleavage of recombinant C terminus of APP in vitro, without specifically modulating the processing of recombinant Notch intracellular domain. These results indicate that FLZ decreases Aβ accumulation in mitochondria by selectively inhibiting γ-secretase. We propose that FLZ is a potential anti-AD drug candidate, and its mechanism may be improving mitochondrial function by reducing the Aβ burden in mitochondria.