Center for Reproductive Medicine, Jilin Medical University, 132013, Jilin, Jilin, China.
Center for Reproductive Medicine, Jilin Medical University, 132013, Jilin, Jilin, China.
Theriogenology. 2025 Jan 15;232:1-8. doi: 10.1016/j.theriogenology.2024.10.033. Epub 2024 Nov 2.
Iron death is a novel type of programmed cell death caused by excessive accumulation of iron-dependent lipid peroxidation products; however, the function of iron death during porcine oocyte maturation and embryo growth is poorly understood. This study was conducted to investigate the mechanism of ferric ammonium citrate (FAC) in regulating iron death in mature oocytes in vitro through the NRF2 signaling pathway, and subsequent embryonic development. The experiment was divided into four groups: 0 (control group), 2, 5, and 10 μM FAC. Western blotting (WB), reactive oxygen species (ROS)assays, mitochondrial membrane potential (MMP) assays, and Quantitative real-time polymerase chain reaction (qRT-PCR) were used to detect the maturation of porcine oocytes in vitro, the protein content of nuclear transcription factor E2-related factor 2 (Nrf2), the distribution of mitochondria, the level of oxidative stress, and the development of embryos fertilized in vitro. The results showed that with increasing FAC concentrations, the oocyte maturation rate in vitro, Nrf2 protein content, MMP, and cleavage rates of in vitro fertilized embryos decreased (significantly in the 5 μM group); the oxidative stress level was significantly increased; the transcript levels of Nrf2, GPX4, and FTH1 mRNAs were significantly decreased; the expression of ACSL4 was significantly upregulated (P < 0.05); and the blastocyst rate of embryos fertilized in vitro was reduced (significantly in the 2 μM group). In conclusion, FAC can regulate Nrf2 protein levels in porcine oocytes matured in vitro to induce iron death, affecting the maturation rate of oocytes, distribution of mitochondria, level of oxidative stress, expression of iron-death-related genes, and development of embryos after in vitro fertilization.
铁死亡是一种新型的程序性细胞死亡，由铁依赖性脂质过氧化产物的过度积累引起；然而，铁死亡在猪卵母细胞成熟和胚胎生长过程中的功能尚不清楚。本研究旨在通过 NRF2 信号通路研究柠檬酸铁铵 (FAC) 在体外调节成熟卵母细胞中铁死亡的机制，以及随后的胚胎发育。实验分为四组：0（对照组）、2、5 和 10 μM FAC。Western blot（WB）、活性氧（ROS）测定、线粒体膜电位（MMP）测定和实时定量聚合酶链反应（qRT-PCR）用于检测体外猪卵母细胞的成熟、核转录因子 E2 相关因子 2（Nrf2）的蛋白含量、线粒体分布、氧化应激水平以及体外受精胚胎的发育。结果表明，随着 FAC 浓度的增加，体外卵母细胞成熟率、Nrf2 蛋白含量、MMP 和体外受精胚胎的卵裂率降低（5 μM 组显著降低）；氧化应激水平显著升高；Nrf2、GPX4 和 FTH1mRNA 的转录水平显著降低；ACSL4 的表达显著上调（P<0.05）；体外受精胚胎的囊胚率降低（2 μM 组显著降低）。综上所述，FAC 可调节体外成熟猪卵母细胞中的 Nrf2 蛋白水平，诱导铁死亡，影响卵母细胞成熟率、线粒体分布、氧化应激水平、铁死亡相关基因的表达以及体外受精后胚胎的发育。
