Melatonin inhibits autophagy in TM3 cells via AKT/FOXO1 pathway

Background Melatonin can regulate apoptosis and autophagy of mouse Leydig cells, but its specific mechanism is still unclear. Methods In this study, we used the TM3 cell line as the research object, and used H2O2 to induce autophagy. After adding 10 ng/ml melatonin, we used qRT-PCR and western-blot to detect autophagy-related gene and protein expression, and flow cytometry to detect cellular ROS level. Results The results showed that melatonin can significantly inhibit the occurrence of autophagy, accompanied by a significant decrease in the expression of Becn1, LC3, and FOXO1 ( P < 0.05), a significant increase in the expression of p62 and pAKT ( P < 0.05), and a significant decrease in ROS level ( P < 0.05). After added the inhibitor of AKT perifosine, the effect of melatonin on inhibiting autophagy was reversed. On this basis, we used small RNA interference technology to knock down the expression of FOXO1, and found that there was no significant change of the expression of genes and proteins related to autophagy and ROS level. Conclusions In summary, melatonin can inhibit H2O2-induced autophagy in TM3 cells through the AKT/FOXO1 pathway.