International Journal of Medical Sciences

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17 October 2017

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Int J Med Sci 2017; 14(5):434-443. doi:10.7150/ijms.18427

Research Paper

Dexamethasone Treatment at the Myoblast Stage Enhanced C2C12 Myocyte Differentiation

Der-Sheng Han1, 2, 3✉, Wei-Shiung Yang4,5,6, Tung-Wei Kao7

1. Department of Physical Medicine and Rehabilitation
2. Community and Geriatric Medicine Research Center, National Taiwan University Hospital, BeiHu Branch, Taipei
3. Department of Physical Medicine and Rehabilitation
4. Department of Internal Medicine, National Taiwan University Hospital, Taipei
5. Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei
6. Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei
7. Division of Geriatric Medicine, Department of Family and Community Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.

Abstract

Background: Glucocorticoids induce skeletal muscle atrophy in many clinical situations; however, their hypertrophic and pro-differentiation effects on myotubes have rarely been reported. We hypothesized that dexamethasone (DEX) has a dual effect on muscle differentiation, and aimed to develop a new differentiation protocol for C2C12 cell line.

Methods: Dose- and time-dependent effect of DEX on C2C12 myoblast cell line was analyzed at myoblast and myotube stage, respectively. The level of differentiation was determined by myh1, pax7, atrogin-1, and myostatin mRNA expression and fusion index.

Results: After differentiation and at the myotube stage, DEX treatment has an atrophic effect. Specifically, the myotube was thinner, the expression of atrogin-1 increased, and the protein content of myosin heavy chain decreased. In contrast, when DEX treatment was performed before the onset of differentiation, we observed an increase in myotube diameter and myosin heavy chain levels, and a decrease in the expression of atrogin-1. The ratio of multinuclear myotube cells increased in the DEX treatment group. The optimal treatment concentration and time was 100 μM and 48 h, respectively. Co-treatment with 10 μM DEX and 100 nM insulin further enhanced the process of myotube differentiation.

Discussion: This novel finding contributed to the explanation on the stage-specific mechanism of glucocorticoid-induced myopathy. A new formula for myoblast differentiation, containing both DEX and insulin, is proposed. Further research is required to understand the complete mechanism of DEX-induced muscle hypertrophy.

Keywords: dexamethasone, glucocorticoid, muscle differentiation, myostatin, atrogin-1.

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How to cite this article:
Han DS, Yang WS, Kao TW. Dexamethasone Treatment at the Myoblast Stage Enhanced C2C12 Myocyte Differentiation. Int J Med Sci 2017; 14(5):434-443. doi:10.7150/ijms.18427. Available from http://www.medsci.org/v14p0434.htm