Centre de Recherche en Reproduction, Développement et Santé Intergénérationnelle, Faculté des Sciences de l'Agriculture et de l'Alimentation, Département des Sciences Animales, Université Laval, Québec, QC, G1V 0A6, Canada.
J Ovarian Res. 2021 Dec 20;14(1):178. doi: 10.1186/s13048-021-00934-6.
Maternal metabolic disorders such as obesity and diabetes are detrimental factors that compromise fertility and the success rates of medically assisted procreation procedures. During metabolic stress, adipose tissue is more likely to release free fatty acids (FFA) in the serum resulting in an increase of FFA levels not only in blood, but also in follicular fluid (FF). In humans, high concentrations of palmitic acid and stearic acid reduced granulosa cell survival and were associated with poor cumulus-oocyte complex (COC) morphology. Obesity and high levels of circulating FFA were also causatively linked to hampered insulin sensitivity in cells and compensatory hyperinsulinemia. To provide a global picture of the principal upstream signaling pathways and genomic mechanisms involved in this metabolic context, human granulosa-like tumor cells (KGN) were treated with a combination of palmitic acid, oleic acid, and stearic acid at the higher physiological concentrations found in the follicular fluid of women with a higher body mass index (BMI) (≥ 30.0 kg/m). We also tested a high concentration of insulin alone and in combination with high concentrations of fatty acids. Transcription analysis by RNA-seq with a cut off for fold change of 1.5 and p-value 0.05 resulted in thousands of differentially expressed genes for each treatment. Using analysis software such as Ingenuity Pathway Analysis (IPA), we were able to establish that high concentrations of FFA affected the expression of genes mainly related to glucose and insulin homoeostasis, fatty acid metabolism, as well as steroidogenesis and granulosa cell differentiation processes. The combination of insulin and high concentrations of FFA affected signaling pathways related to apoptosis, inflammation, and oxidative stress. Taken together, our results provided new information on the mechanisms that might be involved in human granulosa cells exposed to high concentrations of FFA and insulin in the contexts of metabolism disorders.
母体代谢紊乱，如肥胖和糖尿病，是影响生育能力和医学辅助生育程序成功率的有害因素。在代谢应激期间，脂肪组织更有可能在血清中释放游离脂肪酸（FFA），导致 FFA 水平不仅在血液中增加，而且在卵泡液（FF）中增加。在人类中，高浓度的棕榈酸和硬脂酸降低了颗粒细胞的存活率，并与不良的卵丘-卵母细胞复合物（COC）形态相关。肥胖和循环 FFA 水平升高也与细胞胰岛素敏感性下降和代偿性高胰岛素血症有关。为了提供一个在这种代谢背景下涉及的主要上游信号通路和基因组机制的整体画面，用在具有更高体重指数（BMI）（≥30.0kg/m）的女性卵泡液中发现的更高生理浓度的棕榈酸、油酸和硬脂酸组合处理人颗粒细胞样肿瘤细胞（KGN）。我们还单独测试了高浓度的胰岛素和高浓度的脂肪酸组合。RNA-seq 的转录分析，使用 1.5 倍折叠变化和 p 值 0.05 的截止值，导致每种处理的数千个差异表达基因。使用 Ingenuity Pathway Analysis（IPA）等分析软件，我们能够确定高浓度的 FFA 主要影响与葡萄糖和胰岛素稳态、脂肪酸代谢以及类固醇生成和颗粒细胞分化过程相关的基因的表达。胰岛素和高浓度 FFA 的组合影响与细胞凋亡、炎症和氧化应激相关的信号通路。总之，我们的结果提供了关于在代谢紊乱情况下人颗粒细胞暴露于高浓度 FFA 和胰岛素的机制的新信息。
