P-TAK1 acts as a switch between myoblast proliferation phase and differentiation phase in mdx mice via regulating HO-1 expression

Abstract

Skeletal muscle transforming growth factor-β-activated kinase 1 (TAK1) continuous excessive phosphorylation was observed in Duchenne muscular dystrophy (DMD) patients and mdx mice. Inhibiting TAK1 phosphorylation ameliorated fibrosis and muscular atrophy, while TAK1 knockout also impaired muscle regeneration. The definite effect and mechanism of p-TAK1 in muscle regeneration disorder is still obscure. In this study, BaCl2-induced acute muscle injury model was used to investigate the role of p-TAK1 in myoblast proliferation and differentiation phase. The results showed that TAK1 phosphorylation was significantly up-regulated in proliferation phase along with Keap1/Nrf2/HO-1 signaling pathway activation, which was down-regulated in differentiation phase yet. In C2C12 cells, inhibiting TAK1 phosphorylation markedly suppressed the expression of heme oxygenase-1 (HO-1), and both myoblast proliferation and differentiation were inhibited. As for activation, p-TAK1 promoted myoblast proliferation via up-regulating HO-1 level. However, excessive TAK1 phosphorylation (induced by 20 ng·mL−1 TGF-β1) notably up-regulated HO-1 expression, inhibiting myogenic differentiation antigen (MyOD) and myogenic differentiation. A mild p-TAK1 level (induced by 5 or 10 ng·mL−1 TGF-β1) was beneficial for myoblast differentiation. In mdx mice, robust myoblast proliferation and differentiation arrest were observed with high p-TAK1 level in skeletal muscle. HO-1 expression was significantly up-regulated. TAK1 phosphorylation inhibitor NG25 (N-[4-[(4-ethylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl]-4-methyl-3-(1H-pyrrolo[2,3-b]pyridin-4-yloxy)benzamide) significantly inhibited HO-1 expression, relieved excessive myoblast proliferation and differentiation arrest, promoted new myofiber formation, and eventually improved muscle function. In conclusion, p-TAK1 acted as “a switch” between proliferation and differentiation phase. Mitigating p-TAK1 level transformed myoblast excessive proliferation phase into differentiation phase in mdx mouse via regulating HO-1 expression.