A Newly Synthesized Rhamnoside Derivative Alleviates Alzheimer’s Amyloid-β-Induced Oxidative Stress, Mitochondrial Dysfunction, and Cell Senescence through Upregulating SIRT3

Figure 4

PL171 promoted SIRT3 levels and its activity. (a) SK-N-SH cells were incubated with PL171 at different dosages as indicated for 24 h. Mitochondria were isolated and the lysates were prepared for the measurement of total acetylation of mitochondrial proteins using the anti-acetylation antibody (Ac-K) by western blotting. ATP5A was used as a loading control. (b) SK-N-SH cells were treated with 30 μM PL171 at different time points as indicated. Mitochondrial lysates were prepared for the detection of mitochondrial protein acetylation using the anti-acetylation antibody (Ac-K) by western blotting. ATP5A was used as a loading control. (c) After the incubation with PL171 at indicated concentrations for 24 h, mitochondrial lysates were prepared and analyzed using western blotting with indicated antibodies. (d–f) Quantification of relative acetylated MnSOD (AcK68-MnSOD/MnSOD, d), relative acetylated OSCP (AcK139-OSCP/OSCP, e), and relative SIRT3 level (SIRT3/ATP5A, f). (g) Representative blot showing the acetylation of MnSOD and OSCP in the mitochondrial lysates of SK-N-SH cells, challenged of PL171 (30 μM) with or without SIRT3 inhibitor (3-TYP, 20 μM) for 24 h. (h) Quantification of relative MnSOD acetylation in (g). (i) Quantification of relative OSCP acetylation in (g). (j) Representative blot presenting the acetylation of MnSOD in the mitochondrial lysates of SK-N-SH cells pretreated with 30 μM PL171 for 4 h followed by stimulation with 10 μM Aβ₄₂O for 24 h. (k) Quantification of relative MnSOD acetylation in (j). The data are presented as , independent experiments, , , analyzed by one-way ANOVA followed by Bonferroni’s test.

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