Two 18‐norspirostane steroidal saponins as novel mitophagy enhancers improve Alzheimer's disease

Furthermore, the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) results indicated that DTCA&ETCA enhanced the viability of transfected neuronal cells (Figure S7). [...]the flow cytometry analysis revealed that DTCA&ETCA not only decreased GFP intensity but also reduced the proportion of propidium iodide (PI)-positive cells (Figure S8). The transmission electron microscopy images revealed increased engulfment of mitochondria (mt) by autophagosomes (av) in HT-22 cells treated with DTCA, ETCA, or urolithin A (UA, a positive mitophagy activator), suggesting the potential induction of mitophagy by DTCA&ETCA (Figure 2G). Cotreatment with compound C (CC) and SBI0206965 (SBI) attenuated LC3 conversion and GFP-LC3 puncta induced by DTCA&ETCA (Figure S12). [...]SBI significantly reversed the elevated colocalization of GFP-LC3 with mitochondria and the reduced GFP/RFP ratio in DTCA&ETCA-treated HT-22 cells transfected with Mito-QC, a construct expressing a mCherry-GFP tag attached to the outer mitochodrial membrane FIS1 (residues 101-152) (Figure 2I-L). Subsequently, CL2331, CL4176 and BR5270 strains were employed to investigate the anti-AD effects of DTCA&ETCA.8 Figure 3G,H demonstrated that DTCAT&ETCA significantly reduced Aβ aggregations in the anterior region of CL2331worms. [...]in CL4176 worms, DTCA&ETCA exhibited a delay in Aβ-induced paralysis (Figure 3G,I), while in BR5270 worms, it alleviated the food-searching deficit (Figure 3J).