Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPAR mediated mechanism

Tom Ashmore; Lee D. Roberts; Andrea J. Morash; Aleksandra O. Kotwica; John Finnerty; James A. West; Steven A. Murfitt; Bernadette O. Fernandez; Cristina Branco; Andrew S. Cowburn; Kieran Clarke; Randall S. Johnson; Martin Feelisch; Julian L. Griffin; Andrew J. Murray

doi:10.1186/s12915-015-0221-6

Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPAR mediated mechanism

Tom Ashmore, Lee D. Roberts, Andrea J. Morash, Aleksandra O. Kotwica, John Finnerty, James A. West, Steven A. Murfitt, Bernadette O. Fernandez, Cristina Branco, Andrew S. Cowburn, Kieran Clarke, Randall S. Johnson, Martin Feelisch, Julian L. Griffin, Andrew J. Murray^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

39 Citations (Scopus)

38 Downloads (Pure)

Abstract

Background: Insulin sensitivity in skeletal muscle is associated with metabolic flexibility, including a high capacity to increase fatty acid (FA) oxidation in response to increased lipid supply. Lipid overload, however, can result in incomplete FA oxidation and accumulation of potentially harmful intermediates where mitochondrial tricarboxylic acid cycle capacity cannot keep pace with rates of β-oxidation. Enhancement of muscle FA oxidation in combination with mitochondrial biogenesis is therefore emerging as a strategy to treat metabolic disease. Dietary inorganic nitrate was recently shown to reverse aspects of the metabolic syndrome in rodents by as yet incompletely defined mechanisms. Results: Herein, we report that nitrate enhances skeletal muscle FA oxidation in rodents in a dose-dependent manner. We show that nitrate induces FA oxidation through a soluble guanylate cyclase (sGC)/cGMP-mediated PPARβ/δ- and PPARaα-dependent mechanism. Enhanced PPARβ/δ and PPARaα expression and DNA binding induces expression of FA oxidation enzymes, increasing muscle carnitine and lowering tissue malonyl-CoA concentrations, thereby supporting intra-mitochondrial pathways of FA oxidation and enhancing mitochondrial respiration. At higher doses, nitrate induces mitochondrial biogenesis, further increasing FA oxidation and lowering long-chain FA concentrations. Meanwhile, nitrate did not affect mitochondrial FA oxidation in PPARaα^-/- mice. In C2C12 myotubes, nitrate increased expression of the PPARaα targets Cpt1b, Acadl, Hadh and Ucp3, and enhanced oxidative phosphorylation rates with palmitoyl-carnitine; however, these changes in gene expression and respiration were prevented by inhibition of either sGC or protein kinase G. Elevation of cGMP, via the inhibition of phosphodiesterase 5 by sildenafil, also increased expression of Cpt1b, Acadl and Ucp3, as well as CPT1B protein levels, and further enhanced the effect of nitrate supplementation. Conclusions: Nitrate may therefore be effective in the treatment of metabolic disease by inducing FA oxidation in muscle.

Original language	English
Article number	110
Number of pages	17
Journal	BMC Biology
Volume	13
DOIs	https://doi.org/10.1186/s12915-015-0221-6
Publication status	Published - 22 Dec 2015
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2015 Ashmore et al.

Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.

Keywords

Fatty acid oxidation
Metabolism
Mitochondria
Muscle
Nitrate
Nitric oxide

ASJC Scopus subject areas

Biotechnology
Structural Biology
Ecology, Evolution, Behavior and Systematics
Physiology
General Biochemistry,Genetics and Molecular Biology
General Agricultural and Biological Sciences
Plant Science
Developmental Biology
Cell Biology

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Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPARmediated mechanism
Copyright 2015 the authors. This is an open access article published under a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited
Final published version, 1.24 MBLicence: CC BY

Cite this

Ashmore, T., Roberts, L. D., Morash, A. J., Kotwica, A. O., Finnerty, J., West, J. A., Murfitt, S. A., Fernandez, B. O., Branco, C., Cowburn, A. S., Clarke, K., Johnson, R. S., Feelisch, M., Griffin, J. L., & Murray, A. J. (2015). Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPAR mediated mechanism. BMC Biology, 13, Article 110. https://doi.org/10.1186/s12915-015-0221-6

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abstract = "Background: Insulin sensitivity in skeletal muscle is associated with metabolic flexibility, including a high capacity to increase fatty acid (FA) oxidation in response to increased lipid supply. Lipid overload, however, can result in incomplete FA oxidation and accumulation of potentially harmful intermediates where mitochondrial tricarboxylic acid cycle capacity cannot keep pace with rates of β-oxidation. Enhancement of muscle FA oxidation in combination with mitochondrial biogenesis is therefore emerging as a strategy to treat metabolic disease. Dietary inorganic nitrate was recently shown to reverse aspects of the metabolic syndrome in rodents by as yet incompletely defined mechanisms. Results: Herein, we report that nitrate enhances skeletal muscle FA oxidation in rodents in a dose-dependent manner. We show that nitrate induces FA oxidation through a soluble guanylate cyclase (sGC)/cGMP-mediated PPARβ/δ- and PPARaα-dependent mechanism. Enhanced PPARβ/δ and PPARaα expression and DNA binding induces expression of FA oxidation enzymes, increasing muscle carnitine and lowering tissue malonyl-CoA concentrations, thereby supporting intra-mitochondrial pathways of FA oxidation and enhancing mitochondrial respiration. At higher doses, nitrate induces mitochondrial biogenesis, further increasing FA oxidation and lowering long-chain FA concentrations. Meanwhile, nitrate did not affect mitochondrial FA oxidation in PPARaα-/- mice. In C2C12 myotubes, nitrate increased expression of the PPARaα targets Cpt1b, Acadl, Hadh and Ucp3, and enhanced oxidative phosphorylation rates with palmitoyl-carnitine; however, these changes in gene expression and respiration were prevented by inhibition of either sGC or protein kinase G. Elevation of cGMP, via the inhibition of phosphodiesterase 5 by sildenafil, also increased expression of Cpt1b, Acadl and Ucp3, as well as CPT1B protein levels, and further enhanced the effect of nitrate supplementation. Conclusions: Nitrate may therefore be effective in the treatment of metabolic disease by inducing FA oxidation in muscle.",

keywords = "Fatty acid oxidation, Metabolism, Mitochondria, Muscle, Nitrate, Nitric oxide",

author = "Tom Ashmore and Roberts, {Lee D.} and Morash, {Andrea J.} and Kotwica, {Aleksandra O.} and John Finnerty and West, {James A.} and Murfitt, {Steven A.} and Fernandez, {Bernadette O.} and Cristina Branco and Cowburn, {Andrew S.} and Kieran Clarke and Johnson, {Randall S.} and Martin Feelisch and Griffin, {Julian L.} and Murray, {Andrew J.}",

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T1 - Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPAR mediated mechanism

AU - Ashmore, Tom

AU - Roberts, Lee D.

AU - Morash, Andrea J.

AU - Kotwica, Aleksandra O.

AU - Finnerty, John

AU - West, James A.

AU - Murfitt, Steven A.

AU - Fernandez, Bernadette O.

AU - Branco, Cristina

AU - Cowburn, Andrew S.

AU - Clarke, Kieran

AU - Johnson, Randall S.

AU - Feelisch, Martin

AU - Griffin, Julian L.

AU - Murray, Andrew J.

PY - 2015/12/22

Y1 - 2015/12/22

N2 - Background: Insulin sensitivity in skeletal muscle is associated with metabolic flexibility, including a high capacity to increase fatty acid (FA) oxidation in response to increased lipid supply. Lipid overload, however, can result in incomplete FA oxidation and accumulation of potentially harmful intermediates where mitochondrial tricarboxylic acid cycle capacity cannot keep pace with rates of β-oxidation. Enhancement of muscle FA oxidation in combination with mitochondrial biogenesis is therefore emerging as a strategy to treat metabolic disease. Dietary inorganic nitrate was recently shown to reverse aspects of the metabolic syndrome in rodents by as yet incompletely defined mechanisms. Results: Herein, we report that nitrate enhances skeletal muscle FA oxidation in rodents in a dose-dependent manner. We show that nitrate induces FA oxidation through a soluble guanylate cyclase (sGC)/cGMP-mediated PPARβ/δ- and PPARaα-dependent mechanism. Enhanced PPARβ/δ and PPARaα expression and DNA binding induces expression of FA oxidation enzymes, increasing muscle carnitine and lowering tissue malonyl-CoA concentrations, thereby supporting intra-mitochondrial pathways of FA oxidation and enhancing mitochondrial respiration. At higher doses, nitrate induces mitochondrial biogenesis, further increasing FA oxidation and lowering long-chain FA concentrations. Meanwhile, nitrate did not affect mitochondrial FA oxidation in PPARaα-/- mice. In C2C12 myotubes, nitrate increased expression of the PPARaα targets Cpt1b, Acadl, Hadh and Ucp3, and enhanced oxidative phosphorylation rates with palmitoyl-carnitine; however, these changes in gene expression and respiration were prevented by inhibition of either sGC or protein kinase G. Elevation of cGMP, via the inhibition of phosphodiesterase 5 by sildenafil, also increased expression of Cpt1b, Acadl and Ucp3, as well as CPT1B protein levels, and further enhanced the effect of nitrate supplementation. Conclusions: Nitrate may therefore be effective in the treatment of metabolic disease by inducing FA oxidation in muscle.

AB - Background: Insulin sensitivity in skeletal muscle is associated with metabolic flexibility, including a high capacity to increase fatty acid (FA) oxidation in response to increased lipid supply. Lipid overload, however, can result in incomplete FA oxidation and accumulation of potentially harmful intermediates where mitochondrial tricarboxylic acid cycle capacity cannot keep pace with rates of β-oxidation. Enhancement of muscle FA oxidation in combination with mitochondrial biogenesis is therefore emerging as a strategy to treat metabolic disease. Dietary inorganic nitrate was recently shown to reverse aspects of the metabolic syndrome in rodents by as yet incompletely defined mechanisms. Results: Herein, we report that nitrate enhances skeletal muscle FA oxidation in rodents in a dose-dependent manner. We show that nitrate induces FA oxidation through a soluble guanylate cyclase (sGC)/cGMP-mediated PPARβ/δ- and PPARaα-dependent mechanism. Enhanced PPARβ/δ and PPARaα expression and DNA binding induces expression of FA oxidation enzymes, increasing muscle carnitine and lowering tissue malonyl-CoA concentrations, thereby supporting intra-mitochondrial pathways of FA oxidation and enhancing mitochondrial respiration. At higher doses, nitrate induces mitochondrial biogenesis, further increasing FA oxidation and lowering long-chain FA concentrations. Meanwhile, nitrate did not affect mitochondrial FA oxidation in PPARaα-/- mice. In C2C12 myotubes, nitrate increased expression of the PPARaα targets Cpt1b, Acadl, Hadh and Ucp3, and enhanced oxidative phosphorylation rates with palmitoyl-carnitine; however, these changes in gene expression and respiration were prevented by inhibition of either sGC or protein kinase G. Elevation of cGMP, via the inhibition of phosphodiesterase 5 by sildenafil, also increased expression of Cpt1b, Acadl and Ucp3, as well as CPT1B protein levels, and further enhanced the effect of nitrate supplementation. Conclusions: Nitrate may therefore be effective in the treatment of metabolic disease by inducing FA oxidation in muscle.

KW - Fatty acid oxidation

KW - Metabolism

KW - Mitochondria

KW - Muscle

KW - Nitrate

KW - Nitric oxide

U2 - 10.1186/s12915-015-0221-6

DO - 10.1186/s12915-015-0221-6

M3 - Article

C2 - 26694920

AN - SCOPUS:84951801851

SN - 1741-7007

VL - 13

JO - BMC Biology

JF - BMC Biology

M1 - 110

ER -

Nitrate enhances skeletal muscle fatty acid oxidation via a nitric oxide-cGMP-PPAR mediated mechanism

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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