Cargando…
Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear
OBJECTIVES: Many patients who suffer ACL tears have persistent atrophy and weakness even after rehabilitation. Myostatin is a cytokine that directly induces muscle atrophy, and previous studies using rodent models and patients have demonstrated an up regulation of myostatin after ACL tear. Using a p...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968297/ http://dx.doi.org/10.1177/2325967116S00138 |
_version_ | 1782445635619782656 |
---|---|
author | Wolfe, Caroline Nicole Gumucio, Jonathan P. Grekin, Jeremy Khouri, Roger Karl Bedi, Asheesh Mendias, Christopher |
author_facet | Wolfe, Caroline Nicole Gumucio, Jonathan P. Grekin, Jeremy Khouri, Roger Karl Bedi, Asheesh Mendias, Christopher |
author_sort | Wolfe, Caroline Nicole |
collection | PubMed |
description | OBJECTIVES: Many patients who suffer ACL tears have persistent atrophy and weakness even after rehabilitation. Myostatin is a cytokine that directly induces muscle atrophy, and previous studies using rodent models and patients have demonstrated an up regulation of myostatin after ACL tear. Using a preclinical rat model, our objective was to determine if the use of a bioneutralizing antibody against myostatin could prevent muscle atrophy and weakness after ACL tear. METHODS: This study was approved by our IACUC. The left ACL and was transected in 3mo-old male F344 rats. At the time of surgery rats received a single IP injection of either a bioneutralizing antibody against myostatin (10B3, GlaxoSmithKline) or a sham IgG (E1-82.15, GlaxoSmithKline) at a dose of 10mg/kg. Rats (N=8 per group) were sacrificed and tissue was harvested 7 days or 21 days after tear. Contractile force measurements of the extensor digitorum longus (EDL) were performed. The size of muscle fibers in histological sections from the EDL and distal vastus lateralis were also measured. RNA was isolated from the distal rectus femoris and gene expression analysis was performed with RT-qPCR. The expression of each gene was normalized to b actin, and further normalized to the expression of muscles from control, uninjured rats. A two-way ANOVA (alpha=0.05) followed by Holm Sidak post hoc sorting was used to evaluate the effect of time and treatment on measured parameters. RESULTS: Inhibition of myostatin resulted in an increase in muscle fiber size at the 21D time point compared to all other groups (Table 1). While both the sham antibody and myostatin antibody groups increased maximum isometric force production from 7D to 21D, the myostatin antibody group at 21D had a further increase in force compared to the 21D sham group. The muscle E3 ubiquitin ligases atrogin-1 and muscle ring finger 1 (MuRF-1) are the major rate limiting enzymes in muscle protein degradation, and inhibition of myostatin resulted in decreases in the expression of these genes at 7D (Figure 1A). Additionally, IGF-1Ea and IGF-1Eb which are important growth factors that induce muscle protein synthesis, were elevated in the myostatin antibody groups at the 21D time point. For genes related to fibrosis (Figure 1B), although there was no significant difference in MMP-2, there was a significant decrease of MMP-8 expression in the 21D MSTN group when compared to the 21D sham. There were also increases in the expression of TIMP-1 and 2 in the 21D MSTN group. CONCLUSION: In a preclinical rat model, the targeted inhibition of myostatin protected leg muscles from muscle atrophy and improved force production after ACL tear. While the mechanism of action is not entirely clear, it is possible that the inhibition of myostatin preserves strength by limiting the expression of proteolytic enzymes in the post-acute atrophy phase and increasing protein synthesis in later phases. |
format | Online Article Text |
id | pubmed-4968297 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-49682972016-08-11 Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear Wolfe, Caroline Nicole Gumucio, Jonathan P. Grekin, Jeremy Khouri, Roger Karl Bedi, Asheesh Mendias, Christopher Orthop J Sports Med Article OBJECTIVES: Many patients who suffer ACL tears have persistent atrophy and weakness even after rehabilitation. Myostatin is a cytokine that directly induces muscle atrophy, and previous studies using rodent models and patients have demonstrated an up regulation of myostatin after ACL tear. Using a preclinical rat model, our objective was to determine if the use of a bioneutralizing antibody against myostatin could prevent muscle atrophy and weakness after ACL tear. METHODS: This study was approved by our IACUC. The left ACL and was transected in 3mo-old male F344 rats. At the time of surgery rats received a single IP injection of either a bioneutralizing antibody against myostatin (10B3, GlaxoSmithKline) or a sham IgG (E1-82.15, GlaxoSmithKline) at a dose of 10mg/kg. Rats (N=8 per group) were sacrificed and tissue was harvested 7 days or 21 days after tear. Contractile force measurements of the extensor digitorum longus (EDL) were performed. The size of muscle fibers in histological sections from the EDL and distal vastus lateralis were also measured. RNA was isolated from the distal rectus femoris and gene expression analysis was performed with RT-qPCR. The expression of each gene was normalized to b actin, and further normalized to the expression of muscles from control, uninjured rats. A two-way ANOVA (alpha=0.05) followed by Holm Sidak post hoc sorting was used to evaluate the effect of time and treatment on measured parameters. RESULTS: Inhibition of myostatin resulted in an increase in muscle fiber size at the 21D time point compared to all other groups (Table 1). While both the sham antibody and myostatin antibody groups increased maximum isometric force production from 7D to 21D, the myostatin antibody group at 21D had a further increase in force compared to the 21D sham group. The muscle E3 ubiquitin ligases atrogin-1 and muscle ring finger 1 (MuRF-1) are the major rate limiting enzymes in muscle protein degradation, and inhibition of myostatin resulted in decreases in the expression of these genes at 7D (Figure 1A). Additionally, IGF-1Ea and IGF-1Eb which are important growth factors that induce muscle protein synthesis, were elevated in the myostatin antibody groups at the 21D time point. For genes related to fibrosis (Figure 1B), although there was no significant difference in MMP-2, there was a significant decrease of MMP-8 expression in the 21D MSTN group when compared to the 21D sham. There were also increases in the expression of TIMP-1 and 2 in the 21D MSTN group. CONCLUSION: In a preclinical rat model, the targeted inhibition of myostatin protected leg muscles from muscle atrophy and improved force production after ACL tear. While the mechanism of action is not entirely clear, it is possible that the inhibition of myostatin preserves strength by limiting the expression of proteolytic enzymes in the post-acute atrophy phase and increasing protein synthesis in later phases. SAGE Publications 2016-07-29 /pmc/articles/PMC4968297/ http://dx.doi.org/10.1177/2325967116S00138 Text en © The Author(s) 2016 http://creativecommons.org/licenses/by-nc-nd/3.0/ This open-access article is published and distributed under the Creative Commons Attribution - NonCommercial - No Derivatives License (http://creativecommons.org/licenses/by-nc-nd/3.0/), which permits the noncommercial use, distribution, and reproduction of the article in any medium, provided the original author and source are credited. You may not alter, transform, or build upon this article without the permission of the Author(s). For reprints and permission queries, please visit SAGE’s Web site at http://www.sagepub.com/journalsPermissions.nav. |
spellingShingle | Article Wolfe, Caroline Nicole Gumucio, Jonathan P. Grekin, Jeremy Khouri, Roger Karl Bedi, Asheesh Mendias, Christopher Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear |
title | Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear |
title_full | Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear |
title_fullStr | Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear |
title_full_unstemmed | Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear |
title_short | Pharmacological Inhibition of Myostatin Protects Against Atrophy and Weakness after ACL Tear |
title_sort | pharmacological inhibition of myostatin protects against atrophy and weakness after acl tear |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968297/ http://dx.doi.org/10.1177/2325967116S00138 |
work_keys_str_mv | AT wolfecarolinenicole pharmacologicalinhibitionofmyostatinprotectsagainstatrophyandweaknessafteracltear AT gumuciojonathanp pharmacologicalinhibitionofmyostatinprotectsagainstatrophyandweaknessafteracltear AT grekinjeremy pharmacologicalinhibitionofmyostatinprotectsagainstatrophyandweaknessafteracltear AT khourirogerkarl pharmacologicalinhibitionofmyostatinprotectsagainstatrophyandweaknessafteracltear AT bediasheesh pharmacologicalinhibitionofmyostatinprotectsagainstatrophyandweaknessafteracltear AT mendiaschristopher pharmacologicalinhibitionofmyostatinprotectsagainstatrophyandweaknessafteracltear |