Ibuprofen results in alterations of human fetal testis development

Millissia Ben Maamar,Karen R Kilcoyne,Nathalie Dejucq-Rainsford,David M Kristensen,Laurianne Lesné,Isabelle Coiffec,Bernard Jégou,Jean-Philippe Antignac,Christèle Desdoits-Lethimonier,Vincent Lavoué,Cécile Chevrier,Bruno Le Bizec,Kristin Hennig,Rod T Mitchell,Antoine D Rolland,Séverine Mazaud-Guittot

doi:10.1038/srep44184

Abstract

Among pregnant women ibuprofen is one of the most frequently used pharmaceutical compounds with up to 28% reporting use. Regardless of this, it remains unknown whether ibuprofen could act as an endocrine disruptor as reported for fellow analgesics paracetamol and aspirin. To investigate this, we exposed human fetal testes (7–17 gestational weeks (GW)) to ibuprofen using ex vivo culture and xenograft systems. Ibuprofen suppressed testosterone and Leydig cell hormone INSL3 during culture of 8–9 GW fetal testes with concomitant reduction in expression of the steroidogenic enzymes CYP11A1, CYP17A1 and HSD17B3, and of INSL3. Testosterone was not suppressed in testes from fetuses younger than 8 GW, older than 10–12 GW, or in second trimester xenografted testes (14–17 GW). Ex vivo, ibuprofen also affected Sertoli cell by suppressing AMH production and mRNA expression of AMH, SOX9, DHH, and COL2A1. While PGE2 production was suppressed by ibuprofen, PGD2 production was not. Germ cell transcripts POU5F1, TFAP2C, LIN28A, ALPP and KIT were also reduced by ibuprofen. We conclude that, at concentrations relevant to human exposure and within a particular narrow ‘early window’ of sensitivity within first trimester, ibuprofen causes direct endocrine disturbances in the human fetal testis and alteration of the germ cell biology.

Highlights

Ibuprofen is the only of the 3 most common over-the-counter painkiller with paracetamol and aspirin, whose endocrine disruptive potential has not yet been investigated ex vivo
Several recent epidemiological studies have reported that the consumption of ibuprofen during the first trimester is associated with an increased risk of cryptorchidism and/or hypospadias[7,8,33]
Since the in utero assessment of ibuprofen’s effects on the development and function of human fetal testes is impossible for obvious ethical reasons, we developed an ex vivo model system which has already proven to be useful for studying the effects of paracetamol, non-steroidal anti-inflammatory drugs (NSAIDs) and environmental chemicals on the human fetal testis during the first trimester[26,40]

Summary

Introduction

Ibuprofen is the only of the 3 most common over-the-counter painkiller with paracetamol and aspirin, whose endocrine disruptive potential has not yet been investigated ex vivo. In addition to cryptorchidism[7], ibuprofen use by pregnant women has been associated with hypospadias[35], another congenital abnormality featuring a midline fusion defect of the male ventral urethra. The latter association was not found in another study[36]. We used a combination of an organotypic culture system (Fetal Gonad Assay; FEGA) based on culturing human fetal testes fragments, and a human fetal testes xenograft system These approaches, have been separately used for the study of endocrine-disrupting substances[26,27,40,41], but never integrated in the same series of experiments. In the FEGA system, after 1 to 3 days of exposure to ibuprofen at [10−7,8,9,10−4] M, we assessed the gross morphology, endocrine function, and gene expression for the main cell types of the first trimester human fetal testis ex vivo, whilst the xenograft system was used to determine the effect of prolonged (7 day) ibuprofen exposure on the endocrine function of the second trimester testis

Methods

Results

Conclusion