Maduramicin arrests myocardial cells at G0/G1 phase of the cell cycle through inhibiting AKT-Cyclin D1 signaling

Maduramicin, a polyether ionophore antibiotic used as an anticoccidial feed additive in poultry, is toxic to animals and humans and can cause heart failure. The present study was initiated to explore the underlying mechanism of toxicity in H9c2 myocardial cells. We observed using cell imaging and counting methods that maduramicin inhibited cell growth in a concentration-dependent manner. Furthermore, MTT assays showed that maduramicin inhibited cell proliferation in a concentration- and time-dependent manner, and was also confirmed by the finding that maduramicin time dependently blocked the incorporation of BrdU into DNA in H9c2 myocardial cells. Further studies revealed that maduramicin induced accumulation of the cells at G0/G1 phase of the cell cycle and concurrently, there was down regulation of expression of Cyclin D1. In addition, exposure to maduramicin pruned phosphorylation of AKT at both T308 and S473 sites. Finally, we found that pre-treatment of H9c2 myocardial cells with AKT activator SC79, attenuated the inhibitory effects of maduramicin on Cyclin D1 expression and cell proliferation. Collectively, our results suggest that maduramicin-suppressed AKT-Cyclin D1 signaling which results in G0/G1 phase cell cycle arrest, leading to the inhibition of myocardial cell proliferation.