The Effects of Caloric Restriction on Mitochondrial Bionenergetics

<p>(A and B) Each box plot shows the distribution of expression levels from 25th to 75th percentile and the lines inside the boxes denote the medians. The whiskers denote the interval between the 10th and 90th percentiles. The filled circles mark the data points outside the 10th and 90th percentiles. (A) Caloric deficit–induced mitochondrial biogenesis in the CR group (35% ± 5%, ^*<i>p</i> = 0.005) and the CREX group (21% ± 4%, ^#<i>p</i> < 0.004), with no change in the control group (2% ± 2%). The y-axis represents the relative change from baseline in mtDNA for each study group. (B) Analysis of mitochondrial enzyme activity; β-HAD (β-oxidation); CS (TCA cycle), and COX (electron transport chain). The y-axis represents the relative change from baseline in mitochondrial enzyme activity for each study group. (C) Linear correlation between the change from baseline in <i>SIRT1</i> and <i>PPARGC1A</i> mRNAs from baseline in control (○), <i>r</i> = 0.83, <i>p</i> < 0.05; CR (□), <i>r</i> = 0.95, <i>p</i> < 0.01; and CREX participants (▵), <i>r</i> = 0.76, <i>p</i> < 0.05). The linear correlation between the change in <i>SIRT1</i> mRNA and <i>PPARGC1A</i> mRNA from baseline in the CR group (□) remained significant after exclusion of the outlier (<i>r</i> = 0.81, <i>p</i> < 0.01). Changes from baseline to month 6 were analyzed by analysis of variance with baseline values included as covariates.</p

Statistical Dot Plots Showing the Effects of DETA-NO and Adiponectin Treatment on Mitochondrial Content and SIRT1 Protein in Primary Human Myotubes

<div><p>(A–C) Effects of 96 h of 50 μM DETA-NO treatment on mitochondrial content (using MitoTracker Green, <i>p</i> = 0.002) (A), electron transport chain activity (COX, <i>p</i> = 0.018) (B), and mitochondrial membrane potential (TMRE, <i>n</i> = 6, <i>p</i> = 0.042) (C). Treatment effect was determined using independent sample t-test. OD, optical density.</p> <p>(D) Effects of 50 μM DETA-NO on SIRT1 and β-actin protein (top blots); effects of 0.5 μg/ml of globular adiponectin (gAD) and adiponectin receptor R1- and R2-siRNA on SIRT1 and β-actin protein expression (bottom blots). Immunoblotting was undertaken in three participants and data are shown as a representative blot. Means are denoted by the solid black bars.</p></div

Changes in Skeletal Muscle Gene Expression for Key Mitochondrial Proteins

<div><p>(A) TFAM: ^*CR, <i>p</i> = 0.001; ^#CREX, <i>p</i> = 0.014.</p> <p>(B) PPARGC1A: ^*CR, <i>p</i> = 0.004; ^#CREX, <i>p</i> = 0.002.</p> <p>(C) SIRT1: ^*CR, <i>p</i> = 0.016; ^#CREX, <i>p</i> = 0.023.</p> <p>(D) eNOS: ^*CR, <i>p</i> = 0.002; ^#CREX, <i>p</i> = 0.039.</p> <p>Graphs show six-month changes in enzyme expression in response to each intervention. The y-axis represents the relative gene expression change from baseline for each study group. Each box plot shows the distribution of expression levels from 25th to 75th percentile, and the lines inside the boxes denote the medians. The whiskers denote the interval between the 10th and 90th percentiles. The filled circles mark the data points outside the 10th and 90th percentiles. Molecular analysis was performed in 11 of 12 volunteers per group from whom there was sufficient pre- and postintervention sample for this determination. Changes from baseline to month 6 were analyzed by analysis of variance with baseline values included as covariates.</p></div