CellBio  Vol.7 No.2 , June 2018
Agaro-Oligosaccharides Prevent Myostatin Hyperexpression and Myosin Heavy Chain Protein Degradation in C2C12 Myotubes Induced by Tumor Necrosis Factor-α
Abstract: Myostatin is a major factor involved in the regulation of skeletal muscle protein mass. High myostatin levels have been associated with an increase in myotube shrinkage. Enhanced myostatin expression is caused by pro-catabolic reactions involving compounds such as tumor necrosis factor (TNF)-α. The present study investigated the effects of agaro-oligosaccharides (AOSs) on hypercatabolism of myotubes exposed to TNF-α. C2C12 myotubes exposed to TNF-α in the presence or absence of AOSs. Myotube exposure to TNF-α resulted in a reduction in the amount of myosin heavy chain (MyHC) protein and a decrease in myotube diameter, which was associated with increased myostatin mRNA expression. AOSs prevented TNF-α-induced MyHC protein loss and restored normal myostatin mRNA levels, with agarobiose and agarotetraose effectively suppressing the hyperexpression of the mRNA. In addition, expression levels of the known myostatin inhibitors, latent transforming growth factor beta binding protein 3 (Ltbp3) and growth and differentiation factor-associated serum protein 1 (Gasp1) mRNAs, decreased more in TNF-α-induced myotubes than in the TNF-α-free control, possibly resulting in myostatin upregulation. However, AOSs restored nearly normal expression levels of Ltbp3 and Gasp1 mRNA, potentially suppressing myostatin expression. These findings suggest that AOSs could prevent myotube shrinkage induced by TNF-α.
Cite this paper: Shirai, I. , Sakai, T. , Shiba, K. , Uzuhashi, Y. and Karasawa, K. (2018) Agaro-Oligosaccharides Prevent Myostatin Hyperexpression and Myosin Heavy Chain Protein Degradation in C2C12 Myotubes Induced by Tumor Necrosis Factor-α. CellBio, 7, 23-34. doi: 10.4236/cellbio.2018.72003.

[1]   Attaix, D., Baracos, V.E. and Pichard, C. (2012) Muscle Wasting: a Crosstalk between Protein Synthesis and Breakdown Signalling. Current Opinion in Clinical Nutrition and Metabolic Care, 15, 209-210.

[2]   Bonaldo, P. and Sandri, M. (2013) Cellular and Molecular Mechanisms of Muscle Atrophy. Disease Models and Mechanisms, 6, 25-39.

[3]   Rodriguez, J., Vernus, B., Chelh, I., Cassar-Malek, I., Gabillard, J.C., Hadj Sassi, A., Seiliez, I., Picard, B. and Bonnieu, A. (2014) Myostatin and the Skeletal Muscle Atrophy and Hypertrophy Signaling Pathways. Cellular and Molecular Life Sciences, 71, 4361-4371.

[4]   Wang, M., Yu, H., Kim, Y.S., Bidwell, C.A. and, Kuang, S. (2012) Myostatin Facilitates Slow and Inhibits Fast Myosin Heavy Chain Expression during Myogenic Differentiation. Biochemical and Biophysical Research Communications, 426, 83-88.

[5]   McPherron, A.C. and Lee, S.J. (1997) Double Muscling in Cattle due to Mutations in the Myostatin Gene. Proceedings of National Academy of Sciences of the United State of America, 94, 12457-12461.

[6]   Lee, S.J. and McPherron, A.C. (1999) Myostatin and the Control of Skeletal Muscle Mass: Commentary. Current Opinion in Genetics and Development, 9, 604-607.

[7]   Schuelke, M., Wagner, K.R., Stolz, L.E., Hübner, C., Riebel, T., Kömen, W., Braun, T., Tobin, J.F. and Lee, S.J. (2004) Myostatin Mutation Associated with Gross Muscle Hypertrophy in a Child. The New England Journal of Medicine, 351, 2682-2688.

[8]   Enoki, T., Okuda, S., Kudo, Y., Takashima, F., Sagawa, H. and Kato, I. (2010) Oligosaccharides from Agar Inhibit Pro-Inflammatory Mediator Release by Inducing Heme Oxygenase 1. Bioscience, Biotechnology, and Biochemistry, 74, 766-770.

[9]   Higashimura, Y., Naito, Y., Takagi, T., Tanimura, Y., Mizushima, K., Harusato, A., Fukui, A., Yoriki, H., Handa, O., Ohnogi, H. and Yoshikawa, T. (2014) Preventive Effect of Agaro-Oligosaccharides on Non-Steroidal Anti-Inflammatory Drug-Induced Small Intestinal Injury in Mice. Journal of Gastroenterology and Hepatology, 29, 310-317.

[10]   Ajisaka, K., Agawa, S., Nagumo, S., Kurato, K., Yokoyama, T., Arai, K. and Miyazaki, T. (2009) Evaluation and Comparison of the Antioxidative Potency of Various Carbohydrates Using Different Methods. Journal of Agricultural and Food Chemistry, 57, 3102-3107.

[11]   Mirza, K.A., Pereira, S.L., Edens, N.K. and Tisdale, M.J. (2014) Attenuation of Muscle Wasting in Murine C2C12 Myotubes by Epigallocatechin-3-Gallate. Journal of Cachexia, Sarcopenia and Muscle, 5, 339-345.

[12]   Wang, D.T., Yin, Y., Yang, Y.J., Lv, P.J., Shi, Y., Lu, L. and Wei, L.B. (2014) Resveratrol Prevents TNF-α-Induced Muscle Atrophy via Regulation of Akt/mTOR/FoxO1 Signaling in C2C12 Myotubes. International Immunopharmacology, 19, 206-213.

[13]   Shiota, C., Abe, T., Kawai, N., Ohno, A., Teshima-Kondo, S., Mori, H., Terao, J., Tanaka, E. and Nikawa, T. (2015) Flavones Inhibit LPS-Induced Atrogin-1/MAFbx Expression in Mouse C2C12 Skeletal Myotubes. Journal of Nutritional Science and Vitaminology, 61, 188-194.

[14]   Kazlowski, B., Pan, C.L. and Ko, Y.T. (2008) Separation and Quantification of Neoagaro- and Agaro-Oligosaccharide Products Generated from Agarose Digestion by β-agarase and HCl in Liquid Chromatography Systems. Carbohydrate Research, 343, 2443-2450.

[15]   Karasawa, K., Uzuhashi, Y., Hirota, M. and, Otani, H. (2011) A Matured Fruit Extract of Date Palm Tree (Phoenix dactylifera L.) Stimulates the Cellular Immune System in Mice. Journal of Agricultural and Food Chemistry, 59, 11287-11293.

[16]   Williamson, D.L., Butler, D.C. and Alway, S.E. (2009) AMPK Inhibits Myoblast Differentiation through a PGC-1α-Dependent Mechanism. American Journal of Physiology-Endocrinology and Metabolism, 297, E304-E314.

[17]   Sedmak, J.J. and Grossberg, S.E. (1977) A Rapid, Sensitive, and Versatile Assay for Protein Using Coomassie Brilliant Blue G250. Analytical Biochemistry, 79, 544-552.

[18]   Zammit, P.S., Partridge, T.A. and Yablonka-Reuveni, Z. (2006) The Skeletal Muscle Satellite Cell: The Stem Cell That Came in from the Cold. Journal of Histochemistry and Cytochemistry, 54, 1177-1191.

[19]   Chen, H.M. and Yan, X.J. (2005) Antioxidant Activities of Agaro-Oligosaccharides with Different Degrees of Polymerization in Cell-Based System. Biochimica et Biophysica Acta—General Subjects, 1722, 103-111.

[20]   Costelli, P. and Baccino, F.M. (2003) Mechanisms of Skeletal Muscle Depletion in Wasting Syndromes: Role of ATP-Ubiquitin-Dependent Proteolysis. Current Opinion in Clinical Nutrition and Metabolic Care, 6, 407-412.

[21]   Lenk, K., Schur, R., Linke, A., Erbs, S., Matsumoto, Y., Adams, V. and Schuler, G. (2009) Impact of Exercise Training on Myostatin Expression in the Myocardium and Skeletal Muscle in a Chronic Heart Failure Model. European Journal of Heart Failure, 11, 342-348.

[22]   Elkina, Y., von Haehling, S., Anker, S.D. and Springer, J. (2011) The Role of Myostatin in Muscle Wasting: An Overview. Journal of Cachexia, Sarcopenia and Muscle, 2, 143-151.

[23]   Anderson, S.B., Goldberg, A.L. and Whitman, M. (2008) Identification of a Novel Pool of Extracellular Pro-Myostatin in Skeletal Muscle. Journal of Biological Chemistry, 283, 7027-7035.

[24]   Hill, J.J., Qiu, Y., Hewick, R.M. and Wolfman, N.M.(2003) Regulation of Myostatin in Vivo by Growth and Differentiation Factor-Associated Serum Protein-1: A Novel Protein with Protease Inhibitor and Follistatin Domains. Molecular Endocrinology, 17, 1144-1154.

[25]   Ketkar, S., Rathore, A., Kandhare, A., Lohidasan, S., Bodhankar, S., Paradkar, A. and Mahadik, K. (2015) Alleviating Exercise-Induced Muscular Stress Using Neat and Processed Bee Pollen: Oxidative Markers, Mitochondrial Enzymes, and Myostatin Expression in Rats. Integrative Medicine Research, 4, 147-160.