Moderate hypoxia promotes skeletal muscle cell growth and hypertrophy in C2C12 cells
Koki Sakushima, Maki Yoshikawa, Tomohiko Osaki, Naokazu Miyamoto, Takeshi Hashimoto
Abstract
Although several studies have implied that a hypoxic environment may be a factor that influences muscle hypertrophy, scant attention has been paid to the effect of oxygen molecules on the morphological characteristics of muscle. The purpose of the present study was to examine the effect of semisevere (i.e., 5%) to moderate (i.e., 10% or 15%) hypoxic environments on the morphological characteristics of skeletal muscle and the associated mechanisms. C2C12 skeletal muscle cells were divided into various groups, namely, the normoxia group (20.9% O2) and hypoxia groups (5% O2, 10% O2, and 15% O2), and cell growth and the expression of associated proteins in the hypoxia groups were compared with those in the normoxia group. The myotube diameter and cell differentiation index were determined on day 6 by immunocytochemical analyses. The expression of proteins associated with muscle cell differentiation (MyoD and myogenin) and muscle hypertrophy (mTOR and p70s6K) were analyzed by Western blotting. We found that compared with normoxia, a 5% oxygen environment inhibited differentiation and caused muscle atrophy. However, compared with normoxia, a 10% oxygen environment promoted muscle differentiation, and 10% oxygen and 15% oxygen environments induced muscle hypertrophy. Compared with normoxia, a 10% oxygen environment promoted myogenin and the expression of mTOR, p70s6K, and the metabolic signal AMPK. We concluded that a hypoxic environment, if not too severe, may promote muscle differentiation and hypertrophy by increasing the expression of proteins associated with muscle cell differentiation and hypertrophy.