Shanghai Clinical College, Anhui Medical University, Hefei, People's Republic of China.
Centre for Reproductive Medicine, Shanghai 10th People Hospital of Tongji University, Shanghai, People's Republic of China.
Reprod Sci. 2022 Apr;29(4):1343-1356. doi: 10.1007/s43032-021-00691-3. Epub 2021 Aug 10.
This study evaluated the differences in metabolites between cumulus cells (CCs) and mural granulosa cells (MGCs) from human preovulatory follicles to understand the mechanism of oocyte maturation involving CCs and MGCs. CCs and MGCs were collected from women who were undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment. The differences in morphology were determined by immunofluorescence. The metabolomics of CCs and MGCs was measured by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) followed by quantitative polymerase chain reaction (qPCR) and western blot analysis to further confirm the genes and proteins involved in oocyte maturation. CCs and MGCs were cultured for 48 h in vitro, and the medium was collected for detection of hormone levels. There were minor morphological differences between CCs and MGCs. LC-MS/MS analysis showed that there were differences in 101 metabolites between CCs and MGCs: 7 metabolites were upregulated in CCs, and 94 metabolites were upregulated in MGCs. The metabolites related to cholesterol transport and estradiol production were enriched in CCs, while metabolites related to antiapoptosis were enriched in MGCs. The expression of genes and proteins involved in cholesterol transport (ABCA1, LDLR, and SCARB1) and estradiol production (SULT2B1 and CYP19A1) was significantly higher in CCs, and the expression of genes and proteins involved in antiapoptosis (CRLS1, LPCAT3, and PLA2G4A) was significantly higher in MGCs. The level of estrogen in CCs was significantly higher than that in MGCs, while the progesterone level showed no significant differences. There are differences between the metabolomes of CCs and MGCs. These differences may be involved in the regulation of oocyte maturation.
这项研究评估了人类排卵前卵泡中卵丘细胞 (CCs) 和壁颗粒细胞 (MGCs) 之间代谢物的差异,以了解涉及 CCs 和 MGCs 的卵母细胞成熟机制。CCs 和 MGCs 从接受体外受精 (IVF)/胞质内精子注射 (ICSI) 治疗的女性中收集。通过免疫荧光确定形态差异。通过液相色谱串联质谱 (LC-MS/MS) 测量 CCs 和 MGCs 的代谢组学,然后进行定量聚合酶链反应 (qPCR) 和 Western blot 分析,以进一步确认参与卵母细胞成熟的基因和蛋白质。将 CCs 和 MGCs 在体外培养 48 小时,并收集培养基进行激素水平检测。CCs 和 MGCs 之间存在微小的形态差异。LC-MS/MS 分析显示,CCs 和 MGCs 之间有 101 种代谢物存在差异:CCs 中有 7 种代谢物上调,MGCs 中有 94 种代谢物上调。与胆固醇转运和雌二醇生成相关的代谢物在 CCs 中富集,而与抗凋亡相关的代谢物在 MGCs 中富集。胆固醇转运 (ABCA1、LDLR 和 SCARB1) 和雌二醇生成 (SULT2B1 和 CYP19A1) 相关基因和蛋白质的表达在 CCs 中明显更高,抗凋亡 (CRLS1、LPCAT3 和 PLA2G4A) 相关基因和蛋白质的表达在 MGCs 中明显更高。CCs 中的雌激素水平明显高于 MGCs,而孕激素水平无显著差异。CCs 和 MGCs 的代谢组之间存在差异。这些差异可能参与了卵母细胞成熟的调节。