Department of Pharmacology and Neuroscience, Texas Tech University Health Sciences Center (TTUHSC), Lubbock, TX 79430, USA.
Department of Department of Chemical and Biomolecular Engineering, University of Nebraska, Lincoln, NE 68588, USA.
Biomolecules. 2019 Oct 30;9(11):669. doi: 10.3390/biom9110669.
NRF2 is a redox-sensitive transcription factor that depending on the duration or magnitude of the stress, either translocates to the nucleus (beneficial) or is degraded in the cytosol (harmful). However, the role of NRF2-based mechanism(s) under ethanol (E)-induced developmental toxicity in the placental context remains unknown. Here, we used a rat prenatal model of maternal alcohol stress consisting of intermittent ethanol vapor (IEV) daily from GD11 to GD20 with a 6 h ON/18 h OFF in a vapor chamber and in vitro placental model consisting of HTR-8 trophoblasts exposed to 86 mM of E for either 24 h or 48 h. The role of NRF2 was evaluated through the NRF2-transactivation reporter assay, qRT-PCR, and Western blotting for NRF2 and cell growth-promoting protein, and cell proliferation assay. In utero and in vitro E decreased the nuclear NRF2 content and diminished its transactivation ability along with dysregulation of the proliferation indices, PCNA, CYCLIN-D1, and p21. This was associated with a ~50% reduction in cell proliferation in vitro in trophoblasts. Interestingly, this was found to be partially rescued by ectopic overexpression. These results indicate that ethanol-induced dysregulation of NRF2 coordinately regulates PCNA/CYCLIN-D1/p21 involving growth network, at least partially to set a stage for placental perturbations.
NRF2 是一种氧化还原敏感的转录因子，根据应激的持续时间或强度，它要么易位到细胞核（有益），要么在细胞质中降解（有害）。然而，NRF2 为基础的机制在胎盘环境中乙醇（E）诱导的发育毒性中的作用尚不清楚。在这里，我们使用了一个大鼠产前模型，包括在蒸气室中从 GD11 到 GD20 每天间歇性给予乙醇蒸气（IEV），持续 6 小时 ON/18 小时 OFF，以及体外胎盘模型，包括 HTR-8 滋养细胞暴露于 86mM 的 E 24 小时或 48 小时。通过 NRF2 转录激活报告测定、qRT-PCR 和 Western blot 检测 NRF2 和细胞生长促进蛋白，以及细胞增殖测定来评估 NRF2 的作用。在宫内和体外，E 降低了核 NRF2 含量，并减弱了其转录激活能力，同时还失调了增殖指数、PCNA、CYCLIN-D1 和 p21。这与体外滋养细胞的增殖减少了约 50%有关。有趣的是，这部分可以通过异位过表达得到部分挽救。这些结果表明，乙醇诱导的 NRF2 失调协调调节 PCNA/CYCLIN-D1/p21 涉及生长网络，至少部分为胎盘扰动奠定了基础。
