Research Paper Volume 13, Issue 12 pp 16381—16403

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

Sirt6 mediates acacetin-induced reversal of the downregulation of pLKB1, pAMPK and mitophagy signaling in H9C2 cells by D-galactose. (A) Western blots and relative levels of Sirt6 protein in cells treated without (V, vehicle) or with 0.3, 1 or 3 μM) for 72 h. (B) Western blots and relative levels of Sirt6 in cells treated without (control) or with D-galactose (D-gal, 20 mg/mL) or D-galactose plus acacetin (0.3, 1 or 3 μM) for 72 h. (C) Western blots and relative levels of Sirt6 protein in cells transfected with control siRNA or Sirt6 siRNA in the absence (V, vehicle) or presence of 3 μM acacetin. (D) Western blots and relative levels of pLKB1 in cells treated as in (B). (E) Western blots and relative levels of pAMPK in cells treated as in (B). (F) Western blots and relative levels of pAMPK in cells treated as in (C). (G) Western blots and relative levels of PINK1 in cells treated as in (C). (H) Western blots and relative levels of Parkin in cells treated as in (C). (I) Western blots and relative levels of LC3II/LC3I ratio in cells treated as in (C). (n = 5, *P **P #P ##P

Figure 6. Sirt6 mediates acacetin-induced reversal of the downregulation of pLKB1, pAMPK and mitophagy signaling in H9C2 cells by D-galactose. (A) Western blots and relative levels of Sirt6 protein in cells treated without (V, vehicle) or with 0.3, 1 or 3 μM) for 72 h. (B) Western blots and relative levels of Sirt6 in cells treated without (control) or with D-galactose (D-gal, 20 mg/mL) or D-galactose plus acacetin (0.3, 1 or 3 μM) for 72 h. (C) Western blots and relative levels of Sirt6 protein in cells transfected with control siRNA or Sirt6 siRNA in the absence (V, vehicle) or presence of 3 μM acacetin. (D) Western blots and relative levels of pLKB1 in cells treated as in (B). (E) Western blots and relative levels of pAMPK in cells treated as in (B). (F) Western blots and relative levels of pAMPK in cells treated as in (C). (G) Western blots and relative levels of PINK1 in cells treated as in (C). (H) Western blots and relative levels of Parkin in cells treated as in (C). (I) Western blots and relative levels of LC3II/LC3I ratio in cells treated as in (C). (n = 5, *P < 0.05, **P < 0.01 vs. vehicle, control or vehicle of control siRNA; #P < 0.05, ##P < 0.01 vs. D-gal or control siRNA with acacetin).