褪黑素通过AKT/FOXO1信号通路抑制TM3细胞的自噬。

Melatonin inhibits autophagy in TM3 cells via AKT/FOXO1 pathway.

作者信息

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

机构信息

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

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

出版信息

Mol Biol Rep. 2022 Apr;49(4):2925-2932. doi: 10.1007/s11033-021-07107-0. Epub 2022 Jan 8.

DOI:10.1007/s11033-021-07107-0

PMID:34997871

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.

摘要

背景

褪黑素可调节小鼠睾丸间质细胞的凋亡和自噬，但其具体机制仍不清楚。

方法

在本研究中，我们以TM3细胞系为研究对象，用H2O2诱导自噬。加入10 ng/ml褪黑素后，我们用qRT-PCR和western-blot检测自噬相关基因和蛋白表达，用流式细胞术检测细胞ROS水平。

结果

结果表明，褪黑素可显著抑制自噬的发生，同时Becn1、LC3和FOXO1的表达显著降低（P < 0.05），p62和pAKT的表达显著增加（P < 0.05），ROS水平显著降低（P < 0.05）。加入AKT抑制剂perifosine后，褪黑素抑制自噬的作用被逆转。在此基础上，我们用小RNA干扰技术敲低FOXO1的表达，发现自噬相关基因和蛋白的表达及ROS水平均无明显变化。