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Calorie restriction increases muscle mitochondrial biogenesis in healthy humans.

Civitarese AE, Carling S, Heilbronn LK, Hulver MH, Ukropcova B, Deutsch WA, Smith SR, Ravussin E, CALERIE Pennington Te - PLoS Med. (2007)

Bottom Line: In the controls, 24-h EE was unchanged, but in CR and CREX it was significantly reduced from baseline even after adjustment for the loss of metabolic mass (CR, -135 +/- 42 kcal/d, p = 0.002 and CREX, -117 +/- 52 kcal/d, p = 0.008).DNA damage was reduced from baseline in the CR (-0.56 +/- 0.11 arbitrary units, p = 0.003) and CREX (-0.45 +/- 0.12 arbitrary units, p = 0.011), but not in the controls.In primary cultures of human myotubes, a nitric oxide donor (mimicking eNOS signaling) induced mitochondrial biogenesis but failed to induce SIRT1 protein expression, suggesting that additional factors may regulate SIRT1 content during CR.

View Article: PubMed Central - PubMed

Affiliation: Pennington Biomedical Research Center, Baton Rouge, Louisiana, United States of America. CivitaAE@pbrc.edu

ABSTRACT

Background: Caloric restriction without malnutrition extends life span in a range of organisms including insects and mammals and lowers free radical production by the mitochondria. However, the mechanism responsible for this adaptation are poorly understood.

Methods and findings: The current study was undertaken to examine muscle mitochondrial bioenergetics in response to caloric restriction alone or in combination with exercise in 36 young (36.8 +/- 1.0 y), overweight (body mass index, 27.8 +/- 0.7 kg/m(2)) individuals randomized into one of three groups for a 6-mo intervention: Control, 100% of energy requirements; CR, 25% caloric restriction; and CREX, caloric restriction with exercise (CREX), 12.5% CR + 12.5% increased energy expenditure (EE). In the controls, 24-h EE was unchanged, but in CR and CREX it was significantly reduced from baseline even after adjustment for the loss of metabolic mass (CR, -135 +/- 42 kcal/d, p = 0.002 and CREX, -117 +/- 52 kcal/d, p = 0.008). Participants in the CR and CREX groups had increased expression of genes encoding proteins involved in mitochondrial function such as PPARGC1A, TFAM, eNOS, SIRT1, and PARL (all, p < 0.05). In parallel, mitochondrial DNA content increased by 35% +/- 5% in the CR group (p = 0.005) and 21% +/- 4% in the CREX group (p < 0.004), with no change in the control group (2% +/- 2%). However, the activity of key mitochondrial enzymes of the TCA (tricarboxylic acid) cycle (citrate synthase), beta-oxidation (beta-hydroxyacyl-CoA dehydrogenase), and electron transport chain (cytochrome C oxidase II) was unchanged. DNA damage was reduced from baseline in the CR (-0.56 +/- 0.11 arbitrary units, p = 0.003) and CREX (-0.45 +/- 0.12 arbitrary units, p = 0.011), but not in the controls. In primary cultures of human myotubes, a nitric oxide donor (mimicking eNOS signaling) induced mitochondrial biogenesis but failed to induce SIRT1 protein expression, suggesting that additional factors may regulate SIRT1 content during CR.

Conclusions: The observed increase in muscle mitochondrial DNA in association with a decrease in whole body oxygen consumption and DNA damage suggests that caloric restriction improves mitochondrial function in young non-obese adults.

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Statistical Dot Plots Showing the Effects of DETA-NO and Adiponectin Treatment on Mitochondrial Content and SIRT1 Protein in Primary Human Myotubes(A–C) Effects of 96 h of 50 μM DETA-NO treatment on mitochondrial content (using MitoTracker Green, p = 0.002) (A), electron transport chain activity (COX, p = 0.018) (B), and mitochondrial membrane potential (TMRE, n = 6, p = 0.042) (C). Treatment effect was determined using independent sample t-test. OD, optical density.(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.
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pmed-0040076-g003: Statistical Dot Plots Showing the Effects of DETA-NO and Adiponectin Treatment on Mitochondrial Content and SIRT1 Protein in Primary Human Myotubes(A–C) Effects of 96 h of 50 μM DETA-NO treatment on mitochondrial content (using MitoTracker Green, p = 0.002) (A), electron transport chain activity (COX, p = 0.018) (B), and mitochondrial membrane potential (TMRE, n = 6, p = 0.042) (C). Treatment effect was determined using independent sample t-test. OD, optical density.(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.

Mentions: To determine whether nitric oxide (NO) (mimicking eNOS activity) was associated with mitochondrial biogenesis in human skeletal muscle, we treated primary human myotubes (see Methods for clinical characteristics of the donors) with the NO donor DETA-NO. A 4-d treatment with 50 μM of DETA-NO increased the mRNA expression of TFAM (0.62 ± 0.1 versus 1.3 ± 0.1, p < 0.001) and induced mitochondrial biogenesis (indicated by MitotrackerGreen, Figure 3A). DETA-NO treatment decreased cytochrome C oxidase II activity (structural component of complex IV; Figure 3B) and increased mitochondrial membrane potential using TMRE (Figure 3C).


Calorie restriction increases muscle mitochondrial biogenesis in healthy humans.

Civitarese AE, Carling S, Heilbronn LK, Hulver MH, Ukropcova B, Deutsch WA, Smith SR, Ravussin E, CALERIE Pennington Te - PLoS Med. (2007)

Statistical Dot Plots Showing the Effects of DETA-NO and Adiponectin Treatment on Mitochondrial Content and SIRT1 Protein in Primary Human Myotubes(A–C) Effects of 96 h of 50 μM DETA-NO treatment on mitochondrial content (using MitoTracker Green, p = 0.002) (A), electron transport chain activity (COX, p = 0.018) (B), and mitochondrial membrane potential (TMRE, n = 6, p = 0.042) (C). Treatment effect was determined using independent sample t-test. OD, optical density.(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.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1808482&req=5

pmed-0040076-g003: Statistical Dot Plots Showing the Effects of DETA-NO and Adiponectin Treatment on Mitochondrial Content and SIRT1 Protein in Primary Human Myotubes(A–C) Effects of 96 h of 50 μM DETA-NO treatment on mitochondrial content (using MitoTracker Green, p = 0.002) (A), electron transport chain activity (COX, p = 0.018) (B), and mitochondrial membrane potential (TMRE, n = 6, p = 0.042) (C). Treatment effect was determined using independent sample t-test. OD, optical density.(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.
Mentions: To determine whether nitric oxide (NO) (mimicking eNOS activity) was associated with mitochondrial biogenesis in human skeletal muscle, we treated primary human myotubes (see Methods for clinical characteristics of the donors) with the NO donor DETA-NO. A 4-d treatment with 50 μM of DETA-NO increased the mRNA expression of TFAM (0.62 ± 0.1 versus 1.3 ± 0.1, p < 0.001) and induced mitochondrial biogenesis (indicated by MitotrackerGreen, Figure 3A). DETA-NO treatment decreased cytochrome C oxidase II activity (structural component of complex IV; Figure 3B) and increased mitochondrial membrane potential using TMRE (Figure 3C).

Bottom Line: In the controls, 24-h EE was unchanged, but in CR and CREX it was significantly reduced from baseline even after adjustment for the loss of metabolic mass (CR, -135 +/- 42 kcal/d, p = 0.002 and CREX, -117 +/- 52 kcal/d, p = 0.008).DNA damage was reduced from baseline in the CR (-0.56 +/- 0.11 arbitrary units, p = 0.003) and CREX (-0.45 +/- 0.12 arbitrary units, p = 0.011), but not in the controls.In primary cultures of human myotubes, a nitric oxide donor (mimicking eNOS signaling) induced mitochondrial biogenesis but failed to induce SIRT1 protein expression, suggesting that additional factors may regulate SIRT1 content during CR.

View Article: PubMed Central - PubMed

Affiliation: Pennington Biomedical Research Center, Baton Rouge, Louisiana, United States of America. CivitaAE@pbrc.edu

ABSTRACT

Background: Caloric restriction without malnutrition extends life span in a range of organisms including insects and mammals and lowers free radical production by the mitochondria. However, the mechanism responsible for this adaptation are poorly understood.

Methods and findings: The current study was undertaken to examine muscle mitochondrial bioenergetics in response to caloric restriction alone or in combination with exercise in 36 young (36.8 +/- 1.0 y), overweight (body mass index, 27.8 +/- 0.7 kg/m(2)) individuals randomized into one of three groups for a 6-mo intervention: Control, 100% of energy requirements; CR, 25% caloric restriction; and CREX, caloric restriction with exercise (CREX), 12.5% CR + 12.5% increased energy expenditure (EE). In the controls, 24-h EE was unchanged, but in CR and CREX it was significantly reduced from baseline even after adjustment for the loss of metabolic mass (CR, -135 +/- 42 kcal/d, p = 0.002 and CREX, -117 +/- 52 kcal/d, p = 0.008). Participants in the CR and CREX groups had increased expression of genes encoding proteins involved in mitochondrial function such as PPARGC1A, TFAM, eNOS, SIRT1, and PARL (all, p < 0.05). In parallel, mitochondrial DNA content increased by 35% +/- 5% in the CR group (p = 0.005) and 21% +/- 4% in the CREX group (p < 0.004), with no change in the control group (2% +/- 2%). However, the activity of key mitochondrial enzymes of the TCA (tricarboxylic acid) cycle (citrate synthase), beta-oxidation (beta-hydroxyacyl-CoA dehydrogenase), and electron transport chain (cytochrome C oxidase II) was unchanged. DNA damage was reduced from baseline in the CR (-0.56 +/- 0.11 arbitrary units, p = 0.003) and CREX (-0.45 +/- 0.12 arbitrary units, p = 0.011), but not in the controls. In primary cultures of human myotubes, a nitric oxide donor (mimicking eNOS signaling) induced mitochondrial biogenesis but failed to induce SIRT1 protein expression, suggesting that additional factors may regulate SIRT1 content during CR.

Conclusions: The observed increase in muscle mitochondrial DNA in association with a decrease in whole body oxygen consumption and DNA damage suggests that caloric restriction improves mitochondrial function in young non-obese adults.

Show MeSH
Related in: MedlinePlus