Effects of 12 Months of Caloric Restriction on Muscle Mitochondrial Function in Healthy Individuals

Abstract

Context: The effects of caloric restriction (CR) on in vivo muscle mitochondrial function in humans are controversial.

Objective: We evaluated muscle mitochondrial function and associated transcriptional profiles in nonobese humans after 12 months of CR.

Design: Individuals from an ancillary study of the CALERIE 2 randomized controlled trial were assessed at baseline and 12 months after a 25% CR or ad libitum (control) diet.

Setting: The study was performed at Pennington Biomedical Research Center in Baton Rouge, LA.

Participants: Study participants included 51 (34 female subjects, 25 to 50 years of age) healthy nonobese individuals randomized to 1 of 2 groups (CR or control).

Intervention: This study included 12 months of a 25% CR or ad libitum (control) diet.

Main outcomes: In vivo mitochondrial function [maximal ATP synthesis rate (ATPmax), ATPflux/O2 (P/O)] was determined by 31P-magnetic resonance spectroscopy and optical spectroscopy, and body composition was determined by dual-energy X-ray absorptiometry. In a subset of individuals, a muscle biopsy was performed for transcriptional profiling via quantitative reverse transcription polymerase chain reaction and microarrays.

Results: Weight, body mass index (BMI), fat, and fat-free mass (P < 0.001 for all) significantly decreased at month 12 after CR vs control. In vivo ATPmax and P/O were unaffected by 12 months of CR. Targeted transcriptional profiling showed no effects on pathways involved in mitochondrial biogenesis, function, or oxidative stress. A subgroup analysis according to baseline P/O demonstrated that a higher (vs lower) P/O was associated with notable improvements in ATPmax and P/O after CR.

Conclusions: In healthy nonobese humans, CR has no effect on muscle mitochondrial function; however, having a "more coupled" (versus "less coupled") phenotype enables CR-induced improvements in muscle mitochondrial function.

Trial registration: ClinicalTrials.gov NCT00427193 NCT02695511.

Figure 1.

Global gene expression profiling. GSEA of Illumina transcription arrays in muscle tissue mRNA was performed in a subset of individuals owing to tissue limitations and/or biopsy refusal. In the Reactome gene set list, 2 sets were significantly different between groups: (A) lipid transport and lipogenesis and (B) NO-second messenger signaling, which were downregulated in the CR group from baseline to month 12 (CR, n = 12; control, n = 10). Heat map of NOS pathway was generated via the ComplexHeatmap library in R Bioconductor (https://github.com/jokergoo/ComplexHeatmap) using a per-gene scaling metric for delta expression changes (see Materials and Methods).

Figure 2.

Subgroup analysis of in vivo mitochondrial function after CR. (A) Maximal ATP synthesis rate (ATPmax), (B) resting ATP flux/O₂ uptake or P/O, (C) resting ATPflux, and (D) O₂ uptake were measured by ³¹P-MRS and optical spectroscopy in a cohort of healthy nonobese individuals separated by their in vivo mitochondrial coupling (P/O) (coupled versus uncoupled) after 12 months of CR (excluding individuals from the control group) (uncoupled, n = 13; coupled, n = 10). *P < 0.05.