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.

Abstract:The thermal stability of individual glucosinolates within five different Brassica vegetables was studied at 100°C for different incubation times up to 120 minutes. Three vegetables that were used in this study were Brassica oleracea (red cabbage, broccoli and Brussels sprouts) and two were Brassica rapa (pak choi and Chinese cabbage). To rule out the influence of enzymatic breakdown, myrosinase was inactivated prior to the thermal treatments. The stability of three glucosinolates that occurred in all five vegetables (gluconapin, glucobrassicin and 4-methoxyglucobrassicin) varied considerably between the different vegetables. The degradation could be modeled by first order kinetics. The rate constants obtained varied between four to twenty fold between the five vegetables. Brussels sprouts showed the highest degradation rates for all three glucosinolates. The two indole glucosinolates were most stable in red cabbage, while gluconapin was most stable in broccoli. These results indicate the possibilities for plant breeding to select for cultivars in which glucosinolates are more stable during processing.

Kinetic modeling was used as a tool to quantitatively estimate glucosinolate thermal degradation rate constants. Literature shows that thermal degradation rates differ in different vegetables. Well-characterized plant material, leaves of broccoli and Chinese kale plants grown in two seasons, was used in the study. It was shown that a first-order reaction is appropriate to model glucosinolate degradation independent from the season. No difference in degradation rate constants of structurally identical glucosinolates was found between broccoli and Chinese kale leaves when grown in the same season. However, glucosinolate degradation rate constants were highly affected by the season (20-80% increase in spring compared to autumn). These results suggest that differences in glucosinolate degradation rate constants can be due to variation in environmental as well as genetic factors. Furthermore, a methodology to estimate rate constants rapidly is provided to enable the analysis of high sample numbers for future studies.