Research Paper Volume 13, Issue 12 pp 16381—16403

Cardiac senescence is alleviated by the natural flavone acacetin via enhancing mitophagy

Effects of acacetin on cellular senescence induced by D-galactose in H9C2 cardiac cells. (A) Representative images of senescence-associated-β-galactosidase (SA-β-gal) staining for senescent cells in H9C2 cardiac cells treated without (control) or with D-galactose (D-gal, 20 mg/mL) in the absence and presence of 3 μM acacetin for 72 h. (B) Percentage of SA-β-gal-positive senescent cell number in cardiac cells treated without (control) or with D-gal (20 mg/mL) in the absence and presence of 0.3, 1 or 3 μM acacetin for 72 h. (C) Flow cytometry graphs for determining activity of fluorescein di-β-D-galactopyranoside (FDG) in cells treated as in (A). (D) Relative FDG level in cardiac cells treated as in (B). (E) Western blot and relative level of p53 protein in cells treated as in (B). (F) Western blot and relative level of p21 in cells treated as in (B). (n = 5, **P #P ##P

Figure 3. Effects of acacetin on cellular senescence induced by D-galactose in H9C2 cardiac cells. (A) Representative images of senescence-associated-β-galactosidase (SA-β-gal) staining for senescent cells in H9C2 cardiac cells treated without (control) or with D-galactose (D-gal, 20 mg/mL) in the absence and presence of 3 μM acacetin for 72 h. (B) Percentage of SA-β-gal-positive senescent cell number in cardiac cells treated without (control) or with D-gal (20 mg/mL) in the absence and presence of 0.3, 1 or 3 μM acacetin for 72 h. (C) Flow cytometry graphs for determining activity of fluorescein di-β-D-galactopyranoside (FDG) in cells treated as in (A). (D) Relative FDG level in cardiac cells treated as in (B). (E) Western blot and relative level of p53 protein in cells treated as in (B). (F) Western blot and relative level of p21 in cells treated as in (B). (n = 5, **P < 0.01 vs. control. #P < 0.05, ##P < 0.01 vs. D-gal).