Simon Violaine, Avet Charlotte, Grange-Messent Valérie, Wargnier Richard, Denoyelle Chantal, Pierre Alice, Dairou Julien, Dupret Jean-Marie, Cohen-Tannoudji Joëlle
Sorbonne Paris Cité, Université Paris-Diderot, Centre National de la Recherche Scientifique (CNRS) Unité Mixte de Recherche (UMR) 8251, Institut National de la Santé et de la Recherche Médicale (INSERM) U1133, Biologie Fonctionnelle et Adaptative, Physiologie de l'axe gonadotrope, Paris 75013, France.
Sorbonne Universités, Université Pierre et Marie Curie UM CR18, CNRS UMR 8246, INSERM U1130, Neuroscience Paris Seine, Neuroplasticité des Comportements de Reproduction, Paris 75005, France.
Endocrinology. 2017 Oct 1;158(10):3200-3211. doi: 10.1210/en.2017-00374.
Secretion of 17-β-estradiol (E2) by human granulosa cells can be disrupted by various environmental toxicants. In the current study, we investigated whether carbon black nanoparticles (CB NPs) affect the steroidogenic activity of cultured human granulosa cells. The human granulosa cell line KGN and granulosa cells from patients undergoing in vitro fertilization were treated with increasing concentrations of CB NPs (1 to 100 µg/mL) together or not with follicle-stimulating hormone (FSH). We observed that CB NPs are internalized in KGN cells without affecting cell viability. CB NPs could be localized in the cytoplasm, within mitochondria and in association with the outer face of the endoplasmic reticulum membrane. In both cell types, CB NPs reduced in a dose-dependent manner the activity of aromatase enzyme, as reflected by a decrease in E2 secretion. A significant decrease was observed in response to CB NPs concentrations from 25 and 50 µg/mL in KGN cell line and primary cultures, respectively. Furthermore, CB NPs decreased aromatase protein levels in both cells and reduced aromatase transcript levels in KGN cells. CB NPs rapidly activated extracellular signal-regulated kinase 1 and 2 in KGN cells and pharmacological inhibition of this signaling pathway using PD 98059 significantly attenuated the inhibitory effects of CB NPs on CYP19A1 gene expression and aromatase activity. CB NPs also inhibited the stimulatory effect of FSH on aromatase expression and activity. Altogether, our study on cultured ovarian granulosa cells reveals that CB NPs decrease estrogens production and highlights possible detrimental effect of these common NPs on female reproductive health.
多种环境毒物可破坏人颗粒细胞分泌17-β-雌二醇（E2）的功能。在本研究中，我们调查了炭黑纳米颗粒（CB NPs）是否会影响培养的人颗粒细胞的类固醇生成活性。用人颗粒细胞系KGN和接受体外受精患者的颗粒细胞，分别用递增浓度（1至100 µg/mL）的CB NPs处理，同时或不同时添加促卵泡激素（FSH）。我们观察到CB NPs可被KGN细胞内化而不影响细胞活力。CB NPs可定位于细胞质、线粒体内以及与内质网膜的外表面相关联。在这两种细胞类型中，CB NPs均以剂量依赖性方式降低芳香化酶的活性，这可通过E2分泌的减少反映出来。在KGN细胞系和原代培养物中，分别观察到对25和50 µg/mL的CB NPs浓度有显著降低。此外，CB NPs降低了两种细胞中的芳香化酶蛋白水平，并降低了KGN细胞中的芳香化酶转录水平。CB NPs可快速激活KGN细胞中的细胞外信号调节激酶1和2，使用PD 98059对该信号通路进行药理学抑制可显著减弱CB NPs对CYP19A1基因表达和芳香化酶活性的抑制作用。CB NPs还抑制了FSH对芳香化酶表达和活性的刺激作用。总之，我们对培养的卵巢颗粒细胞的研究表明，CB NPs可降低雌激素的产生，并突出了这些常见纳米颗粒对女性生殖健康可能产生的有害影响。
