MHY2233 Attenuates Replicative Cellular Senescence in Human Endothelial Progenitor Cells via SIRT1 Signaling

Figure 3

Roles of MHY2233 in SIRT1 signaling and rescue of replicative senescence in EPCs: (a) EPCs were cultured with DMSO or MHY2233 (10 nM, 100 nM, or 1 μM) for replicative senescence, and SA-β-gal staining was performed. Representative images of SA-β-gal stain in senescent EPCs. Cells stained green are SA-β-gal-positive cells. (b) Quantification of SA-β-gal-positive cells ( per group). (c) Cells were harvested, and total cell lysates were used in western blot analysis for protein expression of SIRT1 in senescent EPCs upon treatment with different concentrations of MHY2233. (d) After chronic treatment of senescent EPCs with DMSO, 10 nM MHY2233, 100 nM resveratrol, or 100 nM EX527, mRNA and total protein were isolated. Measurement of relative mRNA levels of SIRT1, (e) p16, (f) p21, and (g) p53 was analyzed using qRT-PCR ( per group). (h) Whole cell lysates were subjected to western blot analysis for determining protein levels of FOXO3a, SIRT1, acetylated p53, p16^INK4a, and p21 in senescent EPCs. , , and vs. DMSO (control) by a one-way ANOVA test.

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