Jilin Agricultural University, Xincheng Street 2888, Changchun, 130118, China.
Jilin Agricultural University, Xincheng Street 2888, Changchun, 130118, China.
Environ Pollut. 2022 Sep 15;309:119756. doi: 10.1016/j.envpol.2022.119756. Epub 2022 Jul 12.
Monobutyl phthalate (MBP) is the main metabolite of dibutyl phthalate (DBP) in vivo. MBP has a stable structure, can continuously accumulate in living organisms, and has the potentially to harm animal and human reproductive function. In the ovarian follicle microenvironment, MBP may lead to defects in follicular development and steroid production, abnormal meiotic maturation, impaired ovarian function and other reproductive deficits. In this study, SMART-seq was used to investigate the effects of MBP exposure on the in vitro maturation (IVM) and development of porcine oocytes. The results showed that differentially expressed genes after MBP exposure were enriched in the biological processes cytoskeleton, cell apoptosis, endoplasmic reticulum (ER) and mitochondria. Glycine (Gly) improved the developmental potential of porcine oocytes by regulating mitochondrial and ER function. The effect of Gly in protecting oocytes against MBP-induced damage was studied. The results showed that the addition of Gly significantly decreased the rate of MBP-induced spindle abnormalities, decreased the frequency of MBP-induced mitochondria-associated ER membrane (MAM) interactions, and downregulated the protein and gene expression of the linkage molecules Mitofusin 1 (MFN1) and Mitofusin 2 (MFN2) in the MAM. Additionally, treatment with Gly restored the distribution of the 1,4,5-triphosphate receptor 1 (IPR1) and voltage-dependent anion channel 1 (VDAC1), further decreasing the intracellular free calcium concentration ([Ca]) levels and mitochondrial Ca ([Ca]) , increasing the ER Ca ([Ca]) levels, and thus significantly increasing the ER levels and mitochondrial membrane potential (ΔΨ m). Gly also decreased the levels of reactive oxygen species (ROS) and increased the levels of Glutathione (GSH), oocyte apoptosis-related indicators (Caspase-3 activity and Annexin V) and oocyte apoptosis-related genes (BAX, Caspase 3 and AIFM1). Our results suggest that Gly can ameliorate microtubule cytoskeleton abnormalities and improve oocyte maturation by reducing the defective mitochondrial-ER interactions caused by MBP exposure in vitro.
邻苯二甲酸单丁酯（MBP）是体内邻苯二甲酸二丁酯（DBP）的主要代谢物。MBP 具有稳定的结构，可在生物体内不断积累，并有损害动物和人类生殖功能的潜力。在卵巢卵泡微环境中，MBP 可能导致卵泡发育和类固醇产生缺陷、减数分裂成熟异常、卵巢功能受损等生殖缺陷。在这项研究中，使用 SMART-seq 研究了 MBP 暴露对猪卵母细胞体外成熟（IVM）和发育的影响。结果表明，MBP 暴露后差异表达的基因在生物过程中被富集到细胞骨架、细胞凋亡、内质网（ER）和线粒体。甘氨酸（Gly）通过调节线粒体和 ER 功能改善了猪卵母细胞的发育潜能。研究了 Gly 对保护卵母细胞免受 MBP 诱导损伤的作用。结果表明，添加 Gly 可显著降低 MBP 诱导的纺锤体异常率，降低 MBP 诱导的线粒体相关内质网（MAM）相互作用的频率，并下调 MAM 中的连接分子 Mitofusin 1（MFN1）和 Mitofusin 2（MFN2）的蛋白和基因表达。此外，用 Gly 处理可恢复 1,4,5-三磷酸受体 1（IPR1）和电压依赖性阴离子通道 1（VDAC1）的分布，进一步降低细胞内游离钙浓度 ([Ca]) 水平和线粒体 Ca([Ca])，增加内质网 Ca([Ca])水平，从而显著增加内质网水平和线粒体膜电位（ΔΨ m）。Gly 还降低了活性氧（ROS）的水平并增加了谷胱甘肽（GSH）的水平，降低了卵母细胞凋亡相关指标（Caspase-3 活性和 Annexin V）和卵母细胞凋亡相关基因（BAX、Caspase 3 和 AIFM1）的水平。我们的结果表明，Gly 可以通过减少 MBP 暴露引起的线粒体-ER 相互作用的缺陷，改善微管细胞骨架异常，提高卵母细胞成熟度。
