Table 1.
Clinical and echocardiographic characteristics of heart failure patients.
Table 2.
Mitochondrial proteins differentially regulated in dilated cardiomyopathy vs. controls.
Figure 1.
Mitochondrial protein overexpression in dilated human hearts and relationship between PRDX3 and left ventricular function.
(a–g) The influence of dilated cardiomyopathy on the amount of each representative protein involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and stress response (PRDX3) analyzed using western blotting techniques. As shown, all proteins were significantly increased in the DCM group (n = 17) compared with the CNT group (n = 8). The values from the controls were set to 100. Values were normalized to COX IV and finally to the CNT group. The data are expressed as mean+SEM in arbitrary units (optical density). Images are representative of the results obtained for all of the patients with DCM and the CNT included in the study. (h) Scatter plots showing the relationship between PRDX3 protein levels and left ventricular function, specifically with fractional shortening, left ventricular end systolic diameter, and left ventricular end diastolic diameter. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase; FA, fractional shortening; LVESD, left ventricular end systolic diameter; LVEDD, left ventricular end diastolic diameter. *p value<0.05, **p value<0.01, ***p value<0.0001.
Figure 2.
Mitochondrial protein overexpression in dilated human hearts according to immunofluorescence techniques.
Influence of dilated cardiomyopathy on the amount of each representative protein involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and the stress response (PRDX3). Immunofluorescence of (a) ODPA, (b) ETFD, (c) DLDH, (d) AL4A1, (e) ATPA, (f) EFTU, and (g) PRDX3 were significantly increased in patients with dilated cardiomyopathy compared with the control group. Here we show the nucleus co-stained with DAPI (blue). All of the micrographs are representative of the results obtained in four independent experiments for each group and protein studied, DCM (n = 4) and CNT (n = 4). The bar represents 100 µm. The bar graph shows the relative fluorescence intensity in dilated compared to control hearts. The data are expressed as mean ± SEM. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase. *p value<0.05, **p value<0.01.
Figure 3.
Mitochondrial protein localization and overexpression in dilated human hearts analyzed using transmission electron microscopy.
Influence of dilated cardiomyopathy on the amount and localization of the representative proteins involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and stress response (PRDX3). We observed an increase in immunogold labeling in all proteins studied in DCM hearts. We also confirmed the location of all proteins analyzed and observed a similar distribution of each protein upon comparing pathological with control samples. The bar represents 100 nm. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase.
Figure 4.
Levels of mRNA expression determined by RNAseq.
(a) mRNA levels of the ODPA gene (PDHA1), ETFD gene (ETFDH), AL4A1 gene (ALDH4A1), ATPA gene (ATP5A1), and EFTU gene (TUFM) were increased in the dilated group compared to the control group. The data are expressed as mean ± SEM in mRNA relative expression. (b) The scatter plots show the relationship between the mRNA expression of molecules altered involved in substrate utilization (PDHA1 and ETFDH) and also with other altered protein implicated in metabolic processes (ALDH4A1) and protein synthesis (TUFM). These genes showed a relevant correlation with ATP5A1, which is involved in energy production. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu. *p value<0.05, **p value<0.01.