Ingesting a Combined Carbohydrate and Essential Amino Acid Supplement Compared to a Non-Nutritive Placebo Blunts Mitochondrial Biogenesis-Related Gene Expression after Aerobic Exercise

Lee M Margolis, Nancy E Murphy, Christopher T Carrigan, Holly L McClung, Stefan M Pasiakos, Lee M Margolis, Nancy E Murphy, Christopher T Carrigan, Holly L McClung, Stefan M Pasiakos

Abstract

Background: Whether load carriage (LC), an endurance exercise mode composed of the aerobic component of traditional endurance exercise [e.g., cycle ergometry (CE)] and contractile forces characteristic of resistive-type exercise, modulates acute mitochondrial adaptive responses to endurance exercise and supplemental nutrition [carbohydrate + essential amino acids (CHO+EAA)] is not known. Objective: The aim of this study was to examine the effects of LC and CE, with or without CHO+EAA supplementation, on acute markers of mitochondrial biogenesis. Methods: Twenty-five adults performed 90 min of metabolically matched LC (treadmill walking, wearing a vest equal to 30% of body mass) or CE exercise during which CHO+EAA (46 g carbohydrate and 10 g essential amino acids) or non-nutritive control (CON) drinks were consumed. Muscle biopsy samples were collected at rest (pre-exercise), post-exercise, and after 3 h of recovery to assess citrate synthase activity and the expression of mRNA (reverse transcriptase-quantitative polymerase chain reaction) and protein (Western blot). Results: Citrate synthase and phosphorylated p38 mitogen-activated protein kinase (p38 MAPK)Thr180/Tyr182 were elevated postexercise compared with pre-exercise (time main effect, P < 0.05). Peroxisome proliferator-activated γ-receptor coactivator 1α (PGC-1α) expression was highest after recovery for CE compared with LC (exercise-by-time effect, P < 0.05). Sirtuin 1 (SIRT1) expression postexercise was higher for CON than for CHO+EAA treatments (drink-by-time, P < 0.05). Tumor suppressor p53 (p53), mitochondrial transcription factor A (TFAM), and cytochrome c oxidase subunit IV (COXIV) expression was greater for CON than for CHO+EAA treatments (drink main effect, P < 0.05). PGC-1α and p53 expressions were positively associated (P < 0.05) with TFAM (r = 0.629 and 0.736, respectively) and COXIV (r = 0.465 and 0.461, respectively) expressions. Conclusions: Acute mitochondrial adaptive responses to endurance exercise appear to be largely driven by exogenous nutrition availability. Although CE upregulated PGC-1α expression to a greater extent than LC, downstream signaling was the same between modes, suggesting that LC, in large part, elicits the same acute mitochondrial response as traditional, non-weight-bearing endurance exercise. This trial was registered at clinicaltrials.gov as NCT01714479.

Keywords: COXIV; PGC-1α; SIRT1; TFAM; concurrent exercise; p53.

Figures

FIGURE 1
FIGURE 1
PGC-1α pathway schematic. AMPK, AMP-activated protein kinase; PGC-1α, peroxisome proliferator-activated γ-receptor coactivator 1α p38 MAPK, p38 mitogen-activated protein kinase; p53, tumor suppressor p53; RER, respiratory exchange ratio; SIRT1, sirtuin 1; TFAM, mitochondrial transcription factor A.
FIGURE 2
FIGURE 2
Citrate synthase activity (A) and p-p38 MAPKThr180/Tyr182 and p-AMPKThr172 (B). Representative bands correspond to x axis labels below in panel B. Phosphorylation status was normalized to GAPDH, with data presented as fold changes relative to the pre-exercise time period for each group. Values are means ± SEMs. *Different from pre-exercise, P < 0.05. CE, cycle ergometry; CHO+EAA, carbohydrate plus essential amino acids; CON, control; LC, load carriage; p-AMPK, phosphorylated AMP-activated protein kinase; p-p38 MAPK, phosphorylated p38 mitogen-activated protein kinase; POST, postexercise; PRE, pre-exercise.
FIGURE 3
FIGURE 3
mRNA expressions of SIRT1, PGC-1α, and p53 (A) and TFAM and COXIV (B). Data are presented as fold changes relative to mean pre-exercise values for each group. Values are means ± SEMs. *Different from pre-exercise, P < 0.05. +Different from postexercise, P < 0.05. ‡Different from LC, P < 0.05. †Different from CHO+EAA, P < 0.05. CE, cycle ergometry; CHO+EAA, carbohydrate plus essential amino acids; CON, control; COXIV, cytochrome c oxidase subunit IV; LC, load carriage; PGC-1α, peroxisome proliferator-activated γ-receptor coactivator 1α POST, postexercise; PRE, pre-exercise; p53, tumor suppressor p53; REC, recovery; SIRT1, sirtuin 1; TFAM, mitochondrial transcription factor A.
FIGURE 4
FIGURE 4
Correlation of PGC-1α to TFAM (A) and COXIV (B) expression and of p53 to TFAM (C) and COXIV (D) expression. Data are reported as log2 values for correlations to maintain equal scale between positive and negative values. COXIV, cytochrome c oxidase subunit IV; PGC-1α, peroxisome proliferator-activated γ-receptor coactivator 1α p53, tumor suppressor p53; TFAM, mitochondrial transcription factor A.

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