Ectopic Overexpression of PPARγ2 in the Heart Determines Differences in Hypertrophic Cardiomyopathy After Treatment With Different Thiazolidinediones in a Mouse Model of Diabetes

Figures

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  FIGURE 1. Obesity was associated with cardiac dysfunction and ectopically upregulated PPARγ2 expression in the heart. (A) The ratio of the heart weight to tibia length. (B) and (C) The representative 2D echocardiograph and M-mode image of lean and obese mice. (D) Quantification of ejection fraction (EF) and fractional shortening (FS). (E) Measurements of the left ventricle wall thickness in systole (LVPWs) and diastole (LVPWd). (F) The left ventricle internal diameters in systole (LVIDs) and diastole (LVIDd). (G) Representative PPARγ2 bands of Western blot in heart tissues. (H) Quantified protein levels of PPARγ2 in the heart of the LFD group or obese mice. Mean ± SEM, n = 5–8 mice per group, *p < 0.05, **p < 0.01 vs. LFD.

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  FIGURE 2. Overexpression of PPARγ2 is sufficient to lead to hypertrophy in cardiac myocytes. (A) Measurement of PPARγ2 mRNA levels in cardiomyocytes. (B) Analysis of protein levels with Western blot after overexpressing DDK-PPARγ2 in cardiomyocytes. (C) mRNA levels of the hypertrophic marker gene BNP after overexpressing PPARγ2 in cardiomyocytes. (D) representative images of α-SMA staining with CY3 (red) and nuclei with DAPI (blue) in cardiomyocytes. (E) quantified cell surface areas in cardiomyocytes. CON represents transfection of empty PCMV6 plasmid; OE represents transfection of the PCMV6-PPARγ2 plasmid. Means ± SEM for three independent experiments, *p < 0.05, **p < 0.01 vs. CON.

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  FIGURE 3. Rosiglitazone and CMHX008 displayed different changes in cardiac weight and overview morphology. (A) The body weight of mice treated with rosiglitazone and CMHX008 under LFD and HFD conditions; (B) Heart weight to body weight in LFD and HFD after treatment with rosiglitazone and CMHX008 under LFD and HFD conditions; (C) Macroscopic view of representative H&E–stained transverse sections from heart tissues. Means ± SEM, n = 6–10 mice per group, *p < 0.05, **p < 0.01 vs. L-V; # p < 0.05 vs. H-V.

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  FIGURE 4. Rosiglitazone and CMHX008 ameliorate HFD-induced myocardial function but have different effects on echocardiographical indexes. (A) Representative 2D echocardiograph and, (B) M-mode image of mice treated with CMHX008, rosiglitazone or vehicle; (C) Quantification of ejection fraction (EF), (D) fractional shortening (FS), and (E) the ratio of E to A velocity (E/A); (F) Measurements of the left ventricle internal diameters in systole (LVIDs) and diastole (LVIDd), (G) left ventricle wall thickness in systole (LVPWs) and diastole (LVPWd), and (H) the interventricular septal thickness in systole (IVSs) and diastole (IVSd). Means ± SEM, n = 4–6 mice per group, *p < 0.05 vs. L-V, # p < 0.05 vs. H-V.

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  FIGURE 5. Cardiomyocyte changes following treatment with CMHX008 and rosiglitazone in vitro. (A) Representative images of α-SMA staining with CY3 (red) in cardiomyocytes; Measurement of the mRNA levels of the hypertrophic marker genes BNP (B) and ANP (C) in cardiomyocytes. Means ± SEM, from five independent experiments, *p < 0.05 vs. PPARγ2 (−); # p < 0.05 vs. PPARγ2 (+).

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  FIGURE 6. Cardiomyocyte changes in HFD mice following treatment with CMHX008 and rosiglitazone. (A) Representative WGA staining in the heart from HFD mice following treatment with CMHX008 and rosiglitazone; (B) Quantified cardiomyocyte size; Measurement of the mRNA levels of the hypertrophic marker genes BNP (C), ANP (D), and β-MHC (E) in heart tissues. Means ± SEM, n = 5–8 mice per group, *p < 0.05 vs. L-V, # p < 0.05 vs. H-V.