International Journal of Medical Sciences

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23 October 2017

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Int J Med Sci 2016; 13(3):206-219. doi:10.7150/ijms.13268

Research Paper

Dietary Flaxseed Mitigates Impaired Skeletal Muscle Regeneration: in Vivo, in Vitro and in Silico Studies

Felicia Carotenuto1,2*, Alessandra Costa3,4, Maria Cristina Albertini5, Marco Bruno Luigi Rocchi5, Alexander Rudov5, Dario Coletti6, Marilena Minieri7, Paolo Di Nardo1, Laura Teodori2✉

1. Department of Clinical Sciences and Translational Medicine, University of Rome Tor Vergata, Rome, Italy.
2. Diagnostic & Metrology , FSN-TECFIS-DIM, ENEA, Frascati-Rome, Italy.
3. Department of Surgery, McGowan Institute, University of Pittsburgh Medical Center, Pittsburgh, PA, USA.
4. Fondazione San Raffaele, Ceglie Messapica Italy.
5. Department of Biomolecular Sciences; Urbino University “Carlo Bo”; Urbino, Italy.
6. UMR 8256, UPMC P6, Pierre et Marie Curie University, Department of Biological Adaptation and Aging, Paris Cedex, France.
7. Department of Experimental Medicine and Surgery, University of Rome Tor Vergata , Rome, Italy.
*Visiting Researcher at FSN-TECFIS-DIM, ENEA

Abstract

Background: Diets enriched with n-3 polyunsaturated fatty acids (n-3 PUFAs) have been shown to exert a positive impact on muscle diseases. Flaxseed is one of the richest sources of n-3 PUFA acid α-linolenic acid (ALA). The aim of this study was to assess the effects of flaxseed and ALA in models of skeletal muscle degeneration characterized by high levels of Tumor Necrosis Factor-α (TNF).

Methods: The in vivo studies were carried out on dystrophic hamsters affected by muscle damage associated with high TNF plasma levels and fed with a long-term 30% flaxseed-supplemented diet. Differentiating C2C12 myoblasts treated with TNF and challenged with ALA represented the in vitro model. Skeletal muscle morphology was scrutinized by applying the Principal Component Analysis statistical method. Apoptosis, inflammation and myogenesis were analyzed by immunofluorescence. Finally, an in silico analysis was carried out to predict the possible pathways underlying the effects of n-3 PUFAs.

Results: The flaxseed-enriched diet protected the dystrophic muscle from apoptosis and preserved muscle myogenesis by increasing the myogenin and alpha myosin heavy chain. Moreover, it restored the normal expression pattern of caveolin-3 thereby allowing protein retention at the sarcolemma. ALA reduced TNF-induced apoptosis in differentiating myoblasts and prevented the TNF-induced inhibition of myogenesis, as demonstrated by the increased expression of myogenin, myosin heavy chain and caveolin-3, while promoting myotube fusion. The in silico investigation revealed that FAK pathways may play a central role in the protective effects of ALA on myogenesis.

Conclusions: These findings indicate that flaxseed may exert potent beneficial effects by preserving skeletal muscle regeneration and homeostasis partly through an ALA-mediated action. Thus, dietary flaxseed and ALA may serve as a useful strategy for treating patients with muscle dystrophies.

Keywords: muscle dystrophy, diet, flaxseed, Inflammation, myogenesis, in silico.

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How to cite this article:
Carotenuto F, Costa A, Albertini MC, Rocchi MBL, Rudov A, Coletti D, Minieri M, Di Nardo P, Teodori L. Dietary Flaxseed Mitigates Impaired Skeletal Muscle Regeneration: in Vivo, in Vitro and in Silico Studies. Int J Med Sci 2016; 13(3):206-219. doi:10.7150/ijms.13268. Available from http://www.medsci.org/v13p0206.htm