Effect of Differentiation for Mouse Myoblast C<sub>2</sub>C<sub>12</sub> Cells
against Myostatin expression from Dodamtang

Lee, You-seung; Shin, Yoo-jeong; Park, Jong-hyuk; Kim, Seung-mo; Paek, Kyung-min; Park, Chi-sang

The Journal of Internal Korean Medicine > Volume 29(1); 2008 > Article

Original Article

The Journal of Internal Korean Medicine 2008;29(1): 243-257.

導痰湯이 C₂C₁₂세포주로부터 myostatin발현에 의한 심근에 미치는 영향

이유승¹, 신유정¹, 박종혁¹, 김승모², 백경민¹, 박치상¹

¹대구한의대 심계내과학교실
²대구한의대 내과학교실

Effect of Differentiation for Mouse Myoblast C₂C₁₂ Cells against Myostatin expression from Dodamtang

You-seung Lee¹, Yoo-jeong Shin¹, Jong-hyuk Park¹, Seung-mo Kim², Kyung-min Paek¹, Chi-sang Park¹

¹Dept. of Cardiovascular and Neurologic Disease, College of Oriental Medicine, Daegu-Haany University
²Dept. of Internal Medicine, College of Oriental Medicine, Daegu-Haany University

Correspondence

Chi-sang Park ,Tel: 053-770-2105, Email: 2nae2@hanmail.net

Published online: March 30, 2008.

ABSTRACT

Myostatin, a negative regulator of myogenesis, is shown to function by controlling the proliferation of myoblasts. In this study we show that myostatin is an inhibitor of myoblast differentiation and that this inhibition is mediated through Smad 3. To determine MyoD expression by Dodamtang treatment, we compared the expression pattern of C₂C₁₂ mouse myoblasts that constitutively express myostatin with control cells. In vitro, increasing concentrations of Dodamtang reversibly prevented the myogenic blockage of myoblasts by myostatin expression. ELISA assay, Western and confocal analysis indicated that treatment of Dodamtang to the low serum culture media increased the levels of MyoD leading to the inhibition of myogenic differentiation by myostatin. The stable transfection of C₂C₁₂ myoblasts with myostatin expressing constructs did rescue MyoD-induced myogenic differentiation. Consistent with this, the treatment of Dodamtang rescued the expression of a MyoD in C₂C₁₂ myoblasts treated with myostatin. Taken together, these results suggest that induction of MyoD by Dodamtang inhibits myostatin activity and expression via SMAD3 resulting in the rescue of the myoblasts to differentiate into myotubes. Thus we propose that myostatin action by Dodamtang plays a critical role in myogenic differentiation and that the muscular hyperplasia and hypertrophy seen in animals that blockage of functional myostatin is because of deregulated proliferation and differentiation of myoblasts.

Key words: Dodamtang, myoD, myostatin, myoblast