Melatonin Protects Mouse Type A Spermatogonial Stem Cells against Oxidative Stress via The Mitochondrial Thioredoxin System.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2023-11-28 DOI:10.22074/cellj.2023.2003766.1316

Somayeh Heidarizadi, Zahra Rashidi, Cyrus Jalili, Kamran Mansouri, Iraj Rashidi, Behzad Mahaki, Mohammadreza Gholami

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引用次数: 0

Abstract

Objective: Mitochondrial oxidative stress is an important factor in infertility. The mitochondrial thioredoxin system plays an important role in this condition. N-acetyl-5-methoxy tryptamine (melatonin) plays a role in reducing oxidative stress and apoptosis in spermatogonial stem cells (SSCs). In this study, we explore the probable protective effects of melatonin on the mitochondrial thioredoxin system [thioredoxin 2 (Trx2)/Txnip] in SSCs under oxidative stress.

Materials and methods: In this experimental study, SSCs were co-cultured two-dimensionally (2D) with Sertoli cells in DMEM culture medium that contained 10% fetal bovine serum (FBS), 1% antibiotics, and 10 ng/ml glial cell-derived neurotrophic factor (GDNF) for 30 days. The cultured cells were subsequently divided into four groups: control; melatonin (250 μM, 24 hours); melatonin (250 μM, 24 hours)+hydrogen peroxide (H₂O₂, 50 μM, 24 hours); and H₂O₂ (50 μM, 24 hours). Intracellular reactive oxygen species (ROS) production was determined by flow cytometry. Malondialdehyde (MDA) levels were measured by Fluorometry. The expressions of apoptotic and antioxidant genes and nuclear factor erythroid 2-related factor 2 (Nrf2), Trx2, and nicotinamide nucleotide transhydrogenase (NNT) proteins were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Adenosine triphosphate (ATP) levels were measured by fluorometry.

Results: Melatonin reduced H2O2-induced ROS levels and apoptosis in the SSCs. Melatonin also increased mRNA expression of Nrf2, Trx2, NNT, Sirtuin 3 (Sirt3), and decreased mRNA expression of Txnip, and increased protein expressions of Nrf2, Trx2, NNT thereby increasing activity of the mitochondrial thioredoxin system. In addition, melatonin increased ATP levels.

Conclusion: Melatonin increased Trx2 expression through the Nrf2 pathway. This study suggests that melatonin may protect SSCs from oxidative stress in diseases related to infertility.

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褪黑素通过线粒体硫氧还蛋白系统保护小鼠 A 型精原干细胞免受氧化应激影响

目的：线粒体氧化应激是导致不孕症的重要因素：线粒体氧化应激是导致不孕症的一个重要因素。线粒体硫氧还蛋白系统在这种情况下发挥着重要作用。N-乙酰-5-甲氧基色胺（褪黑激素）在减少精原干细胞（SSCs）的氧化应激和凋亡方面发挥作用。本研究探讨了褪黑素对氧化应激下精原干细胞线粒体硫代氧化酶系统[硫代氧化酶2（Trx2）/Txnip]的可能保护作用：在本实验研究中，SSCs与Sertoli细胞在含有10%胎牛血清（FBS）、1%抗生素和10 ng/ml胶质细胞源性神经营养因子（GDNF）的DMEM培养基中进行二维（2D）共培养30天。随后将培养细胞分为四组：对照组；褪黑素（250 μM，24 小时）组；褪黑素（250 μM，24 小时）+过氧化氢（H2O2，50 μM，24 小时）组；H2O2（50 μM，24 小时）组。细胞内活性氧（ROS）的产生通过流式细胞仪测定。丙二醛（MDA）水平用荧光测定法测量。通过实时定量聚合酶链反应（qRT-PCR）和 Western 印迹法测定凋亡和抗氧化基因以及核因子红细胞 2 相关因子 2（Nrf2）、Trx2 和烟酰胺核苷酸转氢酶（NNT）蛋白的表达。荧光测定法测量三磷酸腺苷（ATP）的水平：结果：褪黑素降低了 H2O2 诱导的 ROS 水平，减少了 SSCs 的细胞凋亡。褪黑素还增加了 Nrf2、Trx2、NNT 和 Sirtuin 3 (Sirt3) 的 mRNA 表达，降低了 Txnip 的 mRNA 表达，增加了 Nrf2、Trx2、NNT 的蛋白表达，从而提高了线粒体硫代毒素系统的活性。此外，褪黑素还能提高 ATP 水平：结论：褪黑激素通过 Nrf2 途径增加了 Trx2 的表达。这项研究表明，褪黑素可保护不孕症相关疾病中的间充质干细胞免受氧化应激。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.