Activation of Nrf2 might reduce oxidative stress in human granulosa cells
Activation of Nrf2 might reduce oxidative stress in human granulosa cells
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DOI：
10.1016/j.mce.2017.10.002
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发表时间：
2018-07-15
影响因子：
4.1
通讯作者：
Osuga, Yutaka
中科院分区：
文献类型：
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作者：
Akino, Nana;Wada-Hiraike, Osamu;Osuga, Yutaka
核因子-E2相关因子2（Nrf2）/Kelch样ECH相关蛋白1（Keap1）-抗氧化反应元件（ARE）信号通路是对抗氧化应激（OS）最重要的防御机制之一。有充分证据表明，OS 的平衡状态在人类生殖医学中发挥着重要作用，而 Nrf2 在卵巢颗粒细胞 (GC) 中的生理作用尚未确定。本研究旨在研究Nrf2在GC中的功能。采用免疫组化方法对人卵巢组织中Nrf2和Keap1进行定位，并检测Nrf2和Keap1在人GC中的表达。分离培养人黄素化GC，并向GC中添加Nrf2激活剂过氧化氢（H2O2）或富马酸二甲酯（DMF），通过定量RT-PCR分析Nrf2与抗氧化剂之间的关系。 H2O2 升高了 Nrf2、过氧化氢酶、超氧化物歧化酶 1 (SOD1) 和 8-氧鸟嘌呤 DNA 糖基化酶 (OGG1) 的 mRNA 水平，DMF 处理通过激活 Nrf2 显示出类似但显着的效果。为了确定 Nrf2 与抗氧化剂生成的关系，使用 siRNA 并进行定量 RT-PCR。 Nrf2 表达减少导致这些抗氧化剂 mRNA 水平降低。 H2O2 和/或 DMF 处理后，通过 8-羟基-2'-脱氧鸟苷和荧光染料 2',7'-二氯二氢荧光素二乙酸酯的荧光研究细胞内 ROS 水平，DMF 处理可猝灭 H2O2 产生的细胞内 ROS。这些结果表明，Nrf2 的激活可能会导致人 GC 中 OS 的减轻， 这可以为克服与年龄相关的生育力下降提供新的见解，这种下降主要归因于异常操作系统的积累。 (C) 2017 Elsevier B.V. 保留所有权利。
Nuclear factor-E2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1)-antioxidant response element (ARE) signaling pathway is one of the most important defense mechanisms against oxidative stress (OS). It is well documented that equilibration status of OS plays fundamental roles in human reproductive medicine, and the physiological role of Nrf2 in ovarian granulosa cells (GCs) has not been determined yet. Herein we aimed to study the function of Nrf2 in GCs.Human ovarian tissues were subjected to immunohistochemistry to localize Nrf2 and Keap1 and we detected the expression of Nrf2 and Keap1 in the human GCs. Human luteinized GCs were isolated and cultured, and hydrogen peroxide (H2O2) or Dimethylfumarates (DMF), an activator of Nrf2, were added to GCs to analyze the relationship between Nrf2 and antioxidants by quantitative RT-PCR. The mRNA levels of Nrf2, catalase, superoxide dismutase 1 (SOD1), and 8-Oxoguanine DNA glycosylase (OGG1) were elevated by H2O2, and DMF treatment showed similar but pronounced effects through activation of Nrf2. To determine the relationship of Nrf2 and the generation of antioxidants, siRNAs were used and quantitative RT-PCR were conducted. Decreased expression of Nrf2 resulted in a decreased level of these antioxidant mRNA. Intracellular levels of ROS were investigated by fluorescence of 8-hydroxy-2'-deoxyguanosine and fluorescent dye, 2',7'-dichlorodihydrofluorescein diacetate after H2O2 and/or DMF treatment, and DMF treatment quenched intracellular ROS generation by H2O2.These results show that activation of Nrf2 might lead to alleviate OS in human GCs, and this could provide novel insight to conquer the age-related fertility decline that is mainly attributed to the accumulation of aberrant OS. (C) 2017 Elsevier B.V. All rights reserved.