Carbon Black Nanoparticles Inhibit Aromatase Expression and Estradiol Secretion in Human Granulosa Cells Through the ERK1/2 Pathway

• Published in: Endocrinology (Philadelphia) 2017-10, Vol.158 (10), p.3200-3211
• Main Authors: Simon, Violaine, Avet, Charlotte, Grange-Messent, Valérie, Wargnier, Richard, Denoyelle, Chantal, Pierre, Alice, Dairou, Julien, Dupret, Jean-Marie, Cohen-Tannoudji, Joëlle
• Format: Article
• Language: English
• Subjects: 17β-Estradiol; Aromatase; Aromatase - genetics; Aromatase - metabolism; Aromatase Inhibitors - pharmacology; Black carbon; Carbon; Carbon black; Cell Line; Cell viability; Cytoplasm; Dose-Response Relationship, Drug; Down-Regulation - drug effects; Endocrine Disruptors - pharmacology; Endocrinology; Endoplasmic reticulum; Estradiol - biosynthesis; Estradiol - secretion; Estrogen Antagonists; Estrogens; Extracellular signal-regulated kinase; Female; Fertilization in Vitro; Follicle Stimulating Hormone - administration & dosage; Follicle-stimulating hormone; Gene expression; Granulosa cells; Granulosa Cells - chemistry; Granulosa Cells - enzymology; Granulosa Cells - secretion; Health care; Humans; In vitro fertilization; Kinases; Life Sciences; MAP Kinase Signaling System - drug effects; MAP Kinase Signaling System - physiology; Mitochondria; Nanoparticles; Nanoparticles - administration & dosage; Nanoparticles - analysis; Pharmacology; Reproductive health; Secretion; Sex hormones; Signal transduction; Signaling; Soot - administration & dosage; Soot - analysis; Soot - pharmacology; Toxicants; Transcription
• ISSN: 0013-7227; 1945-7170; 1945-7170
• 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.We show that exposure of human granulosa cells to carbon nanoparticles decreased basal and FSH-induced estradiol secretion and aromatase expression.