Sex and age differences in AMPK phosphorylation, mitochondrial homeostasis, and inflammation in hearts from inflammatory cardiomyopathy patients

Figures

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  FIGURE 1. Alterations in the expression of Sirt1 and AMPK. Statistics from western blot analysis of (a and b) Sirt1, (c and d) total AMPK, and (e and f) phosphorylated AMPK (Thr172) performed with lysates of cardiac tissue from diseased and non‐diseased, younger, and older men and women. Data are shown as the means ± SEM (n = 3‐10/group). Mann–Whitney Test; *p < 0.05 versus corresponding control. All data were normalized to the corresponding control and expressed in relative units (r.u.).

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  FIGURE 2. Alterations in the expression of PGC‐1α and mitochondrial proteins. Statistics from western blot expression analysis of (a and b) PGC‐1α, (c and d) TOM40, and (e and f) TIM23, performed with lysates of cardiac tissue from diseased and non‐diseased, younger and older men and women. (g and h) Relative mRNA expression of NRF1 and ERRα in diseased and non‐diseased cardiac tissue in older men and women. Data are shown as the means ± SEM (n = 3‐9/group). Mann–Whitney Test; *p < 0.05 versus corresponding control. All data were normalized to the corresponding control and expressed in relative units (r.u.).

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  FIGURE 3. Alterations in the expression of mitochondrial genes and mitochondrial mass. (a–f) Relative mRNA expression of mitochondria‐ and nucleus‐encoded genes in diseased and non‐diseased cardiac tissue in younger and older men and women. (g‐h) Mitochondrial mass was analyzed as the mt‐RNR2/β‐globin ratio. Data are shown as the means ± SEM (n = 3‐8/group). Mann–Whitney Test; *p < 0.05 versus corresponding control. All data were normalized to the corresponding control.

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  FIGURE 4. Alterations in the expression of autophagy markers. Statistics from expression analyses of (a and b) ATG5, (c and d) SQSTM1 at the mRNA and protein levels, (e) LC3II/LC3I ratio, and (f) LAMP2 performed with lysates of cardiac tissue from diseased and non‐diseased older men and women. Data are shown as the means ± SEM (n = 3‐8/group). Mann–Whitney Test; *p < 0.05, **p < 0.01, ***p < 0.001 versus corresponding control. All data were normalized to the corresponding control and expressed in relative units (r.u.).

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  FIGURE 5. Alterations in mitochondrial acetylation state and expression of anti‐oxidative enzymes. Statistics from western blot analyses of (a and b) Sirt3, (c and d) acetylated SOD2 (acSOD2), (e and f) total SOD2, and (g and h) catalase performed with lysates of cardiac tissue from diseased and non‐diseased, younger and older men and women. Data are shown as the means ± SEM (n = 3‐5/group). Mann–Whitney Test; *p < 0.05, **p < 0.01 versus corresponding control. All data were normalized to the corresponding control and expressed in relative units (r.u.).

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  FIGURE 6. Alterations in pro‐ and anti‐inflammatory markers. Statistics from western blot analyses of (a) NF‐κB and real‐time PCR analyses of (b) TLR4, (c) IL‐12, (d) IL‐18, and (g) IL‐10 mRNA expression performed with lysates of cardiac tissue from diseased and non‐diseased, younger and older men and women. Data are shown as the means ± SEM (n = 3‐9/group). All data were normalized to the corresponding control and western blot data expressed in relative units (r.u.). (e and f) Representative images and statistics of IL‐18 staining of cardiac tissue from younger and older men and women with DCMI (n = 3‐5/group). Magnification 200×. Scale bar = 100 μm. Mann–Whitney Test; *p < 0.05 versus corresponding control.

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  FIGURE 7. Alterations in the expression of senescence markers. Statistics from western blot analyses of (a) lamin B1, (b) p53, (c) phospho‐H2A.X, and (d) MMP3 and real‐time PCR analyses of (e) VEGF, (f) IL‐6, and (g) TGF‐β mRNA expression performed with lysates of cardiac tissue from diseased and non‐diseased, older men and women. Data are shown as the means ± SEM (n = 3‐9/group). Mann–Whitney Test; *p < 0.05 versus corresponding control. All data were normalized to the corresponding control and expressed in relative units (r.u.). (H) Absolute telomere length measurement performed with DNA of cardiac tissue from diseased and non‐diseased, older men and women. Data are shown as the means ± SEM (n = 4‐7/group).