Arsenic Inhibits Myogenic Differentiation and Muscle Regeneration

BACKGROUND: The incidence of low birth weights is increased in offspring of women who are exposed to high concentrations of arsenic in drinking water compared with other women. We hypothesized that effects of arsenic on birth weight may be related to effects on myogenic differentiation. OBJECTIVE: We investigated the effects of arsenic trioxide (As(2)O(3)) on the myogenic differentiation of myoblasts in vitro and muscle regeneration in vivo. METHODS: C2C12 myoblasts and primary mouse and human myoblasts were cultured in differentiation media with or without As(2)O(3) (0.1–0.5 μM) for 4 days. Myogenic differentiation was assessed by myogenin and myosin heavy chain expression and multinucleated myotube formation in vitro; skeletal muscle regeneration was tested using an in vivo mouse model with experimental glycerol myopathy. RESULTS: A submicromolar concentration of As(2)O(3) dose-dependently inhibited myogenic differentiation without apparent effects on cell viability. As(2)O(3) significantly and dose-dependently decreased phosphorylation of Akt and p70s6k proteins during myogenic differentiation. As(2)O(3)-induced inhibition in myotube formation and muscle-specific protein expression was reversed by transfection with the constitutively active form of Akt. Sections of soleus muscles stained with hematoxylin and eosin showed typical changes of injury and regeneration after local glycerol injection in mice. Regeneration of glycerol-injured soleus muscles, myogenin expression, and Akt phosphorylation were suppressed in muscles isolated from As(2)O(3)-treated mice compared with untreated mice. CONCLUSION: Our results suggest that As(2)O(3) inhibits myogenic differentiation by inhibiting Akt-regulated signaling.