Redundancy in regulation of lipid accumulation in skeletal muscle during prolonged fasting in obese men

Morten L Høgild, Anders Gudiksen, Henriette Pilegaard, Hans Stødkilde-Jørgensen, Steen Bønløkke Pedersen, Niels Møller, Jens O L Jørgensen, Niels Jessen, Morten L Høgild, Anders Gudiksen, Henriette Pilegaard, Hans Stødkilde-Jørgensen, Steen Bønløkke Pedersen, Niels Møller, Jens O L Jørgensen, Niels Jessen

Abstract

Fasting in human subjects shifts skeletal muscle metabolism toward lipid utilization and accumulation, including intramyocellular lipid (IMCL) deposition. Growth hormone (GH) secretion amplifies during fasting and promotes lipolysis and lipid oxidation, but it is unknown to which degree lipid deposition and metabolism in skeletal muscle during fasting depends on GH action. To test this, we studied nine obese but otherwise healthy men thrice: (a) in the postabsorptive state ("CTRL"), (b) during 72-hr fasting ("FAST"), and (c) during 72-hr fasting and treatment with a GH antagonist (GHA) ("FAST + GHA"). IMCL was assessed by magnetic resonance spectroscopy (MRS) and blood samples were drawn for plasma metabolomics assessment while muscle biopsies were obtained for measurements of regulators of substrate metabolism. Prolonged fasting was associated with elevated GH levels and a pronounced GHA-independent increase in circulating medium- and long-chain fatty acids, glycerol, and ketone bodies indicating increased supply of lipid intermediates to skeletal muscle. Additionally, fasting was associated with a release of short-, medium-, and long-chain acylcarnitines to the circulation from an increased β-oxidation. This was consistent with a ≈55%-60% decrease in pyruvate dehydrogenase (PDHa) activity. Opposite, IMCL content increased ≈75% with prolonged fasting without an effect of GHA. We suggest that prolonged fasting increases lipid uptake in skeletal muscle and saturates lipid oxidation, both favoring IMCL deposition. This occurs without a detectable effect of GHA on skeletal muscle lipid metabolism.

Trial registration: ClinicalTrials.gov NCT02500095.

Keywords: Growth hormone; fasting; intramyocellular lipid; pyruvate dehydrogenase activity; skeletal muscle.

Conflict of interest statement

The authors have nothing to disclose.

© 2019 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

Figures

Figure 1
Figure 1
CD36 protein content, PDH activity, and PDK4 expression. (a) Geometric mean ± SE for CD36 protein content. (b) Mean ± SE for PDHa activity. (c) Mean ± SE for log mRNA expression of PDK4. N = 9. (d) Stain‐free blot image used for total protein normalization of CD36 (Figure 1a)
Figure 2
Figure 2
mRNA content of PPAR‐regulated genes and PPARα, β, and γ. (a) Mean ± SE for log mRNA content of FABP4. (b) Mean ± SE for log mRNA content of CPT1α. (c) Mean ± SE for log mRNA content of CPT1α. (d) Mean ± SE for log mRNA content of ANGPTL4. (e) Mean ± SE for log mRNA content of UCP2. (f) Mean ± SE for log mRNA content of GLUT4. (g) Mean ± SE for log mRNA content of NRF1. (h) Mean ± SE for log mRNA content of PGC1α. (i) Mean ± SE for log mRNA content of PPARα. (j) Mean ± SE for log mRNA content of PPARβ. (k) Mean ± SE for log mRNA content of PPARγ. 12‐hr fasting (CTRL), 72‐hr fasting (FAST), and 72‐hr fasting + GHA (FAST + GHA). N = 9. Not significant (NS). N = 9
Figure 3
Figure 3
IMCL and EMCL during 72‐hr fasting. (a) Mean ± SE for the relative IMCL content. (b) Mean ± SE for the relative EMCL content. (c) Representative water‐suppressed point‐resolved spectroscopy sequence of the volume of interest in musculus tibialis anterior. (d) Representative image of an oblique‐plane T1‐weighted gradient echo pulse sequences with the voxel positioned in a homogeneous part of the musculus tibialis anterior. 12‐hr fasting (CTRL), 72‐hr fasting (FAST), and 72‐hr fasting + GHA (FAST + GHA). N = 8. Not significant (NS).
Figure 4
Figure 4
Protein content of mitochondrial proteins (a) Geometric mean ± SE for cytochrome c. (b) Geometric mean ± SE for SDHA. (c) Geometric mean ± SE for VDAC. (d) Geometric mean ± SE for PDH‐E1α. 12‐hr fasting (CTRL), 72‐hr fasting (FAST), and 72‐hr fasting + GHA (FAST + GHA). N = 9. Not significant (NS). Representative western blots are for CTRL, FAST, and FAST + GHA are shown below the figures. N = 9. (e) Stain‐free blot image used for total protein normalization of Cytochrom C, SDHA, and VDAC (Figure 4a–c). (f) Stain‐free blot image used for total protein normalization of PDH (Figure 4d)
Figure 5
Figure 5
Plasma metabolite concentrations measured at t = 0 min during 12‐hr fasting, 72‐hr fasting alone, and 72‐hr fasting with concomitant pegvisomant administration. (a–h) Data for each metabolite are presented as box and whiskers plot (maximum value, 75th percentile, median, 25th percentile, minimum value). 12‐hr fasting (CTRL), 72‐hr fasting (FAST), and 72‐hr fasting + GHA (FAST + GHA). N = 9

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