(–)-Epicatechin Modulates Mitochondrial Redox in Vascular Cell Models of Oxidative Stress

Figure 1

(a–d) Mitochondrial and cellular superoxide measurement in both HG and AA perturbations: Superoxide concentrations were measured in HUVECs by electron paramagnetic resonance spectroscopy using two different spin probes to differentiate mitochondrial (Mito-TEMPO-H) and total cell (CMH) superoxide. CM or mito-TEMPO nitroxide radicals concentration was obtained by simulating the spectra using the SpinFit module incorporated in the Xenon software of the bench-top EMXnano EPR spectrometer followed by the SpinCount module (Bruker). Nitroxide concentrations were normalized to total protein. Mitochondrial superoxide (a–c) and MnSOD protein expression (e) was assessed in cells exposed to 0.1 μM and 1.0 μM EPICAT , in control, and HG- and AA-treated cells. Representative spectra is shown (c), and total cellular superoxide during a HG perturbation is shown (d), . For analysis of superoxide, control data from all experiments was pooled , and separate tests were run on each experiment of different EPICAT concentrations. AA experiments, AA effect, both experiments, EPICAT effect for 1.0 μM concentration only (a). HG experiments, glucose effect, EPICAT effect, 1.0 μM concentration only (b), two-way ANOVA, Tukey multiple comparisons analysis. A long horizontal bar over the entire graph indicates an interaction effect, smaller bars over the EPICAT groups indicates an EPICAT effect, while bars with tabs indicate a single main effect of either AA or HG. Post hoc analyses are described as as compared to control, as compared to treatment, as compared to EPICAT control. Data are expressed as .

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