Involvement of Sirtuins and Klotho in Cardioprotective Effects of Exercise Training Against Waterpipe Tobacco Smoking-Induced Heart Dysfunction

FIGURE 1. Effects of waterpipe smoke inhalation on serum cardiac troponin I (A) and cotinine levels (B) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = 7.*P < 0.05; ***P < 0.001 vs. CTL; #P < 0.05, ###P < 0.001 vs. S; &&&P < 0.001 vs. Ex.

FIGURE 2. Effects of waterpipe smoke inhalation on left ventricular systolic pressure (LVSP) (A), left ventricular end diastolic pressure (LVEDP) (B), and heart rate (C) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = 7. *P < 0.05; **P < 0.01; ***P < 0.001 vs. CTL; &&P < 0.01, &&&P<0.001 vs. Ex.

FIGURE 3. Effects of waterpipe smoke inhalation on positive dP/dt max (+dP/dt max) and negative dP/dt max (-dP/dt max) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = 7. **P < 0.01; ***P < 0.001 vs. CTL; ##P < 0.01 vs. S; &&P < 0.01; &&&P < 0.001 vs. Ex.

FIGURE 4. Effects of waterpipe smoke inhalation on contractility index (A) and Tau index (B) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = 7. ***P < 0.001 vs. CTL; ###P < 0.001 vs. S; &&&P < 0.001 vs. Ex.

FIGURE 5. Effects of waterpipe smoke inhalation on sirtuin 1 (SIRT1) levels in serum (A) and in the heart (B), and sirtuin 3 (SIRT3) levels in serum (C) and in the heart (D) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = (6–7). *P < 0.05; ***P < 0.001 vs. CTL; #P < 0.05 vs. S; &P < 0.05; &&P < 0.01; &&&P < 0.001 vs. Ex.

FIGURE 6. Effects of waterpipe smoke inhalation on α-Klotho levels in serum (A) and in the heart (B) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = (6–7). *P < 0.05; ***P < 0.001 vs. CTL; ###P < 0.001 vs. S; &&P < 0.01 vs. Ex.

FIGURE 7. Effects of waterpipe smoke inhalation on Bax levels in serum (A) and in the heart (B), and Bcl-2 levels in serum (C) and in the heart (D) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n = (6–7). *P < 0.05, ***P < 0.001 vs. CTL; #P < 0.05, ###P < 0.001 vs. S; &&P < 0.01, &&&P < 0.001 vs. Ex.

FIGURE 8. Effects of waterpipe smoke inhalation on Bax/Bcl-2 ratio in serum (A) and in the heart (B) in experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. The results are presented as mean ± SEM, n (6–7). *P < 0.05, ***P < 0.001 vs. CTL; #P < 0.05, ###P < 0.001 vs. S; &&&P < 0.001 vs. Ex.

FIGURE 9. Effects of waterpipe smoke inhalation on the expression of cardiac pS9-GSK-3β proteins (A) and pS9-GSK-3β/ GAPDH ratio (B) in different experimental groups. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inahaltion for 8 weeks. The results are presented as mean ± SEM, n = (6–7). *P < 0.05 vs. CTL.

FIGURE 10. Histological changes of the heart in experimental groups. (A) CTL, (B) Ex, (C) S, and (D) Ex + S. Heart tissues were stained with hematoxylin and eosin and visualized under a light microscope. CTL, control group; Ex, group subjected to exercise training for 8 weeks; S, group subjected to waterpipe tobacco smoke inhalation for 8 weeks; Ex + S, group subjected to exercise training and waterpipe tobacco smoke inhalation for 8 weeks. Histopathological examination was performed under light microscopy at a magnification of ×40. Yellow arrow: interstitial edema, red arrow: congestion, brown arrow: cellular degeneration.