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Melatonin inhibits autophagy in TM3 cells via AKT/FOXO1 pathway

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Abstract

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.

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Acknowledgements

The authors thank the funds supported by ‘National Key Research and Development Program of China’ (Grant No. 2018YFE0122000), ‘National Natural Science Foundation of China’ (Grant No. 31772600), ‘Jilin Province Young and Middle-aged Science and Technology Innovation Leading Talent Project’ (Grant No. 20200301031RQ).

Author information

Author notes

  1. Jun Wang & Wenfa Lu

    Present address: College of Animal Science and Technology, Jilin Agricultural University, 2888 Xincheng Street, 130118, Changchun, Jilin Province, China

  2. Zhiqiang Li and Hongtao Wang have contributed equally to this work.

Authors and Affiliations

  1. Joint Laboratory of the Modern Agricultural Technology International Cooperation, Ministry of Education, Jilin Agricultural University, 130118, Changchun, Jilin, China

    Zhiqiang Li, Hongtao Wang, Kaiyan Zhang, Jing Zhao, Hongyu Liu, Xin Ma, Jing Guo, Jun Wang & Wenfa Lu

  2. Key Lab of the Animal Production, Product Quality, and Security, Ministry of Education, Jilin Agricultural University, 130118, Changchun, Jilin, China

    Zhiqiang Li, Hongtao Wang, Kaiyan Zhang, Jing Zhao, Hongyu Liu, Xin Ma, Jing Guo, Jun Wang & Wenfa Lu

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  1. Zhiqiang Li
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Contributions

Conceptualization, WL and JW; Methodology, ZL and JZ; Software, HW, HL; Validation, ZL, XM, and JG; Formal analysis, ZL; Investigation, KZ; Data curation, ZL; Writing—original draft preparation, ZL; Writing—review & editing, WL and JW; Supervision, WL and JW; Project administration, WL; Funding acquisition, WL.

Corresponding authors

Correspondence to Jun Wang or Wenfa Lu.

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Li, Z., Wang, H., Zhang, K. et al. Melatonin inhibits autophagy in TM3 cells via AKT/FOXO1 pathway. Mol Biol Rep 49, 2925–2932 (2022). https://doi.org/10.1007/s11033-021-07107-0

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  • DOI: https://doi.org/10.1007/s11033-021-07107-0

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