Department of Obstetrics, Maternal and Child Health Hospital of Hubei Province, Wuhan City, China.
J Obstet Gynaecol Res. 2021 Nov;47(11):3797-3806. doi: 10.1111/jog.15009. Epub 2021 Sep 15.
This article aims to investigate the possible mechanism by which nuclear factor erythroid 2-related factor 2 (Nrf2) signaling exerts protective effects on the progression of preeclampsia (PE).
HTR-8/SVneo cells were cultured under hypoxic conditions to establish PE cell model. Cell viability and invasion were respectively examined by CCK-8 assay and transwell assay. The detection of oxidative stress and Fe content was carried out by corresponding commercial kits, and the expression of corresponding proteins and genes was detected by western blot and qPCR. After Nrf2 was overexpressed in HTR-8/SVneo cells, the above assays were conducted to confirm the exact effects of Nrf2 activation on ferroptosis.
Hypoxia reduced the invasion of HTR-8/SVneo cells, induced oxidative stress (upregulated levels of glutathione [GSH], malondialdehyde [MDA], and reactive oxygen species [ROS]), and induced ferroptosis which exhibited by upregulated Fe and downregulated expression of solute carrier family 7 member 11/System xCT (SCL7A11), glutathione peroxidase 4 (GPX4), and ferroportin1 (FPN1). Meanwhile, hypoxia promoted the translocation of Nrf2 to the nucleus, leading to Nrf2/HO-1 signaling activation. These changes upon hypoxia induction were strengthened by ferric ammonium citrate (FAC) but were abolished by deferoxamine (DFO). Moreover, Nrf2 overexpression in hypoxia-induced HTR-8/SVneo cells exerted inhibitory effects on levels of GSH, MDA, ROS, and Fe , and promotive effects on Nrf2/HO-1 signaling activation and expression of SCL7A11, GPX4, and FPN1, indicating that Nrf2 overexpression decreased oxidative stress and ferroptosis in hypoxia-induced HTR-8/SVneo cells.
Collectively, activation of Nrf2 signaling alleviated hypoxia-induced invasion lacking, oxidative stress, and ferroptosis in hypoxia-induced HTR-8/SVneo cells, suggesting that Nrf2 signaling activation play a protective role in PE, which can provide instructions for the targeted treatment of PE.
本文旨在探讨核因子红细胞 2 相关因子 2（Nrf2）信号通路对子痫前期（PE）进展发挥保护作用的可能机制。
培养 HTR-8/SVneo 细胞使其处于低氧条件下，建立 PE 细胞模型。通过 CCK-8 检测法和 Transwell 小室侵袭实验分别检测细胞活力和侵袭能力。采用相应的商业试剂盒检测氧化应激和铁含量，采用 Western blot 和 qPCR 检测相应蛋白和基因的表达。在 HTR-8/SVneo 细胞中转染 Nrf2 后，进行上述实验以验证 Nrf2 激活对铁死亡的确切影响。
低氧条件降低了 HTR-8/SVneo 细胞的侵袭能力，诱导氧化应激（GSH、丙二醛 [MDA] 和活性氧 [ROS] 水平升高），并诱导铁死亡，表现为铁水平升高和溶质载体家族 7 成员 11/系统 xCT（SCL7A11）、谷胱甘肽过氧化物酶 4（GPX4）和铁蛋白 1（FPN1）表达下调。同时，低氧诱导 Nrf2 向核内易位，导致 Nrf2/HO-1 信号通路激活。这些低氧诱导的变化通过柠檬酸铁铵（FAC）增强，但被去铁胺（DFO）消除。此外，低氧诱导的 HTR-8/SVneo 细胞中转染 Nrf2 对 GSH、MDA、ROS 和铁水平的抑制作用，以及对 Nrf2/HO-1 信号通路激活和 SCL7A11、GPX4 和 FPN1 表达的促进作用，表明 Nrf2 过表达降低了低氧诱导的 HTR-8/SVneo 细胞中的氧化应激和铁死亡。
总之，Nrf2 信号通路的激活缓解了低氧诱导的 HTR-8/SVneo 细胞侵袭缺失、氧化应激和铁死亡，提示 Nrf2 信号通路激活在 PE 中发挥保护作用，为 PE 的靶向治疗提供了指导。
