Cargando…

L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy

BACKGROUND: The purpose of this study was to examine if L-leucine (Leu), β-hydroxy-β-methylbutyrate (HMB), or creatine monohydrate (Crea) prevented potential atrophic effects of myostatin (MSTN) on differentiated C2C12 myotubes. METHODS: After four days of differentiation, myotubes were treated with...

Descripción completa

Detalles Bibliográficos
Autores principales: Mobley, Christopher Brooks, Fox, Carlton D, Ferguson, Brian S, Amin, Rajesh H, Dalbo, Vincent J, Baier, Shawn, Rathmacher, John A, Wilson, Jacob M, Roberts, Michael D
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4134516/
https://www.ncbi.nlm.nih.gov/pubmed/25132809
http://dx.doi.org/10.1186/1550-2783-11-38
_version_ 1782330882599682048
author Mobley, Christopher Brooks
Fox, Carlton D
Ferguson, Brian S
Amin, Rajesh H
Dalbo, Vincent J
Baier, Shawn
Rathmacher, John A
Wilson, Jacob M
Roberts, Michael D
author_facet Mobley, Christopher Brooks
Fox, Carlton D
Ferguson, Brian S
Amin, Rajesh H
Dalbo, Vincent J
Baier, Shawn
Rathmacher, John A
Wilson, Jacob M
Roberts, Michael D
author_sort Mobley, Christopher Brooks
collection PubMed
description BACKGROUND: The purpose of this study was to examine if L-leucine (Leu), β-hydroxy-β-methylbutyrate (HMB), or creatine monohydrate (Crea) prevented potential atrophic effects of myostatin (MSTN) on differentiated C2C12 myotubes. METHODS: After four days of differentiation, myotubes were treated with MSTN (10 ng/ml) for two additional days and four treatment groups were studied: 1) 3x per day 10 mM Leu, 2) 3x per day 10 mM HMB, 3) 3x per day 10 mM Crea, 4) DM only. Myotubes treated with DM without MSTN were analyzed as the control condition (DM/CTL). Following treatment, cells were analyzed for total protein, DNA content, RNA content, muscle protein synthesis (MPS, SUnSET method), and fiber diameter. Separate batch treatments were analyzed for mRNA expression patterns of myostatin-related genes (Akirin-1/Mighty, Notch-1, Ski, MyoD) as well as atrogenes (MuRF-1, and MAFbx/Atrogin-1). RESULTS: MSTN decreased fiber diameter approximately 30% compared to DM/CTL myotubes (p < 0.001). Leu, HMB and Crea prevented MSTN-induced atrophy. MSTN did not decrease MPS levels compared to DM/CTL myotubes, but MSTN treatment decreased the mRNA expression of Akirin-1/Mighty by 27% (p < 0.001) and MyoD by 26% (p < 0.01) compared to DM/CTL myotubes. shRNA experiments confirmed that Mighty mRNA knockdown reduced myotube size, linking MSTN treatment to atrophy independent of MPS. Remarkably, MSTN + Leu and MSTN + HMB myotubes had similar Akirin-1/Mighty and MyoD mRNA levels compared to DM/CTL myotubes. Furthermore, MSTN + Crea myotubes exhibited a 36% (p < 0.05) and 86% (p < 0.001) increase in Akirin-1/Mighty mRNA compared to DM/CTL and MSTN-only treated myotubes, respectively. CONCLUSIONS: Leu, HMB and Crea may reduce MSTN-induced muscle fiber atrophy by influencing Akirin-1/Mighty mRNA expression patterns. Future studies are needed to examine if Leu, HMB and Crea independently or synergistically affect Akirin-1/Mighty expression, and how Akirin-1/Mighty expression mechanistically relates to skeletal muscle hypertrophy in vivo.
format Online
Article
Text
id pubmed-4134516
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-41345162014-08-17 L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy Mobley, Christopher Brooks Fox, Carlton D Ferguson, Brian S Amin, Rajesh H Dalbo, Vincent J Baier, Shawn Rathmacher, John A Wilson, Jacob M Roberts, Michael D J Int Soc Sports Nutr Research Article BACKGROUND: The purpose of this study was to examine if L-leucine (Leu), β-hydroxy-β-methylbutyrate (HMB), or creatine monohydrate (Crea) prevented potential atrophic effects of myostatin (MSTN) on differentiated C2C12 myotubes. METHODS: After four days of differentiation, myotubes were treated with MSTN (10 ng/ml) for two additional days and four treatment groups were studied: 1) 3x per day 10 mM Leu, 2) 3x per day 10 mM HMB, 3) 3x per day 10 mM Crea, 4) DM only. Myotubes treated with DM without MSTN were analyzed as the control condition (DM/CTL). Following treatment, cells were analyzed for total protein, DNA content, RNA content, muscle protein synthesis (MPS, SUnSET method), and fiber diameter. Separate batch treatments were analyzed for mRNA expression patterns of myostatin-related genes (Akirin-1/Mighty, Notch-1, Ski, MyoD) as well as atrogenes (MuRF-1, and MAFbx/Atrogin-1). RESULTS: MSTN decreased fiber diameter approximately 30% compared to DM/CTL myotubes (p < 0.001). Leu, HMB and Crea prevented MSTN-induced atrophy. MSTN did not decrease MPS levels compared to DM/CTL myotubes, but MSTN treatment decreased the mRNA expression of Akirin-1/Mighty by 27% (p < 0.001) and MyoD by 26% (p < 0.01) compared to DM/CTL myotubes. shRNA experiments confirmed that Mighty mRNA knockdown reduced myotube size, linking MSTN treatment to atrophy independent of MPS. Remarkably, MSTN + Leu and MSTN + HMB myotubes had similar Akirin-1/Mighty and MyoD mRNA levels compared to DM/CTL myotubes. Furthermore, MSTN + Crea myotubes exhibited a 36% (p < 0.05) and 86% (p < 0.001) increase in Akirin-1/Mighty mRNA compared to DM/CTL and MSTN-only treated myotubes, respectively. CONCLUSIONS: Leu, HMB and Crea may reduce MSTN-induced muscle fiber atrophy by influencing Akirin-1/Mighty mRNA expression patterns. Future studies are needed to examine if Leu, HMB and Crea independently or synergistically affect Akirin-1/Mighty expression, and how Akirin-1/Mighty expression mechanistically relates to skeletal muscle hypertrophy in vivo. BioMed Central 2014-08-13 /pmc/articles/PMC4134516/ /pubmed/25132809 http://dx.doi.org/10.1186/1550-2783-11-38 Text en Copyright © 2014 Mobley et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Mobley, Christopher Brooks
Fox, Carlton D
Ferguson, Brian S
Amin, Rajesh H
Dalbo, Vincent J
Baier, Shawn
Rathmacher, John A
Wilson, Jacob M
Roberts, Michael D
L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy
title L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy
title_full L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy
title_fullStr L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy
title_full_unstemmed L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy
title_short L-leucine, beta-hydroxy-beta-methylbutyric acid (HMB) and creatine monohydrate prevent myostatin-induced Akirin-1/Mighty mRNA down-regulation and myotube atrophy
title_sort l-leucine, beta-hydroxy-beta-methylbutyric acid (hmb) and creatine monohydrate prevent myostatin-induced akirin-1/mighty mrna down-regulation and myotube atrophy
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4134516/
https://www.ncbi.nlm.nih.gov/pubmed/25132809
http://dx.doi.org/10.1186/1550-2783-11-38
work_keys_str_mv AT mobleychristopherbrooks lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT foxcarltond lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT fergusonbrians lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT aminrajeshh lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT dalbovincentj lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT baiershawn lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT rathmacherjohna lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT wilsonjacobm lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy
AT robertsmichaeld lleucinebetahydroxybetamethylbutyricacidhmbandcreatinemonohydratepreventmyostatininducedakirin1mightymrnadownregulationandmyotubeatrophy