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. 2015:2015:232836.
doi: 10.1155/2015/232836. Epub 2015 Feb 24.

Direct effects of (-)-epicatechin and procyanidin B2 on the respiration of rat heart mitochondria

Dalia M Kopustinskiene  1 , Arunas Savickas  1 , David Vetchý  2 , Ruta Masteikova  2 , Arturas Kasauskas  3 , Jurga Bernatoniene  1
Affiliations

Direct effects of (-)-epicatechin and procyanidin B2 on the respiration of rat heart mitochondria

Dalia M Kopustinskiene et al. Biomed Res Int. 2015.

Abstract

Flavonol (-)-epicatechin and its derived dimer procyanidin B2, present in high amounts in cocoa products, have been shown to exert beneficial effects on the heart and cardiovascular system; however, their mechanism of action has not been fully elucidated. We studied effects of (-)-epicatechin and procyanidin B2 on the oxidative phosphorylation of isolated rat heart mitochondria. (-)-Epicatechin and procyanidin B2 had stimulating effect (up to 30% compared to control) on substrate-driven (State 2) mitochondrial respiration. Their effect was dependent on the respiratory substrates used. (-)-Epicatechin at higher concentrations (from 0.27 µg/mL) significantly decreased (up to 15%) substrate- and ADP-driven (State 3) mitochondrial respiration in case of pyruvate and malate oxidation only. Procyanidin B2 (0.7-17.9 ng/mL) inhibited State 3 respiration rate up to 19%, the most profound effect being expressed with succinate as the substrate. (-)-Epicatechin at concentrations of 0.23 µg/mL and 0.46 µg/mL prevented loss of the cytochrome c from mitochondria when substrate was succinate, supporting the evidence of membrane stabilizing properties of this flavonol. Thus, both (-)-epicatechin and procyanidin B2 directly influenced mitochondrial functions and the observed effects could help to explain cardiometabolic risk reduction ascribed to the consumption of modest amounts of cocoa products.

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Figures

Figure 1
Figure 1
Chemical structure of (−)-epicatechin (1) and procyanidin B2 (2).
Figure 2
Figure 2
Effect of (−)-epicatechin (a) and procyanidin B2 (b) on the respiration rate of isolated rat heart mitochondria in the State 2. Effect was expressed in % of initial respiration rate, which was 76 ± 2 nmol O/min/mg protein (substrates, pyruvate and malate, P + M), 200 ± 14 nmol O/min/mg protein (substrate succinate in the presence of amytal, S(+A)), and 89 ± 6 nmol O/min/mg protein (substrates, palmitoyl-L-carnitine and malate, Pcarn + M). * P < 0.05 versus control, n = 5. The results were analysed with one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test.
Figure 3
Figure 3
Effect of (−)-epicatechin (a) and procyanidin B2 (b) on the respiration rate of isolated rat heart mitochondria in State 3. Effect was expressed in % of substrates- and ADP-driven respiration rate, which was 455 ± 16 nmol O/min/mg protein (substrates, pyruvate and malate, P + M), 453 ± 7 nmol O/min/mg protein (substrate succinate in the presence of amytal, S(+A)), and 392 ± 10 nmol O/min/mg protein (substrates, palmitoyl-L-carnitine and malate, Pcarn + M). * P < 0.05 versus control, n = 5. The results were analysed with one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test.
Figure 4
Figure 4
Effect of (−)-epicatechin (1) on the cytochrome c release from isolated rat heart mitochondria oxidizing succinate in the presence of amytal. VADP+Cyt  c, cytochrome c (32 μM), stimulated State 3 respiration rate of heart mitochondria, VADP, State 3 respiration rate of heart mitochondria. * P < 0.05 versus control, n = 5. The results were analysed with one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test.
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