Figshare20240613.pdf. Data obtained by American Heart Association Transformational Project Grant, 19TPA34850170.Update

Figure 1. Sirt1 is inactivated by HFD feeding. (A) Sirt1 at protein levels were examined after 1 month of HFD feeding in Sirt1cKO mice, and after 3 months of HFD feeding in wild type mice. (B) Sirt1 at protein level was examined in Sirt1cTG mice. (C) Expression of Sirt1 in 3 months of HFD feeding. (D) Decrease in NAD/NADH ratio by HFD feeding. After 1 month of HFD feeding, NAD and NADH levels were examined. (E-F) HFD-induced protein acetylation and acetylation of p65 at K310 are enhanced in Sirt1cKO (E) but inhibited in Sirt1cTG (F) mice. The numbers of mice examined in each experimental group were: 6 (A), 6-7 (B), 7(C), 8 (D), 7 (acetyl lysine, E), 6-8 (acetyl-RelA at K310, E), 5 (acetyl lysine, F) and 6-8 (acetyl-RelA at K310, E). Statistical significance was determined with Student t test (A and D) and ANOVA (B, E to F).Figure 2. Sirt1 negatively regulates HFD-induced diastolic dysfunction. (A) HFD-induced diastolic dysfunction is exacerbated in Sirt1cKO mice. (B-C) HFD or loss of Sirt1 does not significantly affect cardiac systolic function (B) and cardiac hypertrophy (C). (D) HFD-induced increase in body weight was not significantly changed Sirt1cKO mice. (E) HFD-induced diastolic dysfunction is ameliorated in Sirt1cTG mice. (F-G) HFD or gain of Sirt1 does not significantly affect cardiac systolic function (F) and cardiac hypertrophy (G). (H) HFD-induced increase in body weight was not significantly changed Sirt1cKO mice. PV loop analyses (A and E), echocardiographic measurement (B and F), and heart (C and G) and body weight (D and H) measurements were performed after 3 months of HFD feeding. The numbers of mice examined in each experimental group were: 6-10 (A), 7-8 (B), 14-20 (C and D), 5-8 (E), 7-13 (F), and 10-15 (G and H). Statistical significance was determined with ANOVA (A,B,C,D,E,F, and G) and Kruskall-Wallis (H).Figure 3. Exogenous Sirt1 expression with AAV inhibits HFD-induced diastolic dysfunction, fibrosis and TGFβ1 signaling. After 1 month of HFD feeding, AAV-Sirt1 or AAV-GFP (control) were injected. PV loop analyses (A) and PASR staining (B) were performed after 2 month of AAV injection. The numbers of mice examined in each experimental group were: 5 (A) and 8 (B). Statistical significance was determined with ANOVA.