Figure 1. Ibuprofen and Leydig cell steroidogenic function.

(A–C) Representative images of CYP11A1 immunostaining in explants of a 7.9 gestational week (GW) human fetal testis cultured with DMSO or ibuprofen. CYP11A1 appears brown (3,3′-diaminobenzidine tetrahydrochloride staining), and sections were counterstained with hematoxylin. Scale bar = 100 μm. (D) Testosterone production as a function of culture duration in 7–7.9 GW human fetal testes in the presence of DMSO (Control) or 10⁻⁵M of ibuprofen. Results are expressed as fold change from the respective control testis (% of Control). Values are means ± SEM of 6 testes from 6 different fetuses. Repeated measures analysis of variance (ANOVA) on ranks was performed (p = 0.172). (E–G) Testosterone production after culture of 8-9.9 GW (E,F) and 10–12 GW human fetal testes (G) in the presence of DMSO (Control) or 10⁻⁷–10⁻⁴M of ibuprofen for 72 h (E,G) and for 24, 48 and 72 h (F). Results are expressed as fold change from the first day of culture (FC to D0). Values are means ± SEM of 14–37 testes from 14–37 fetuses for the 8–9.9 GW group, 6–14 testes from 6–14 fetuses for the 8–9.9 GW at 24, 48 and 72 h group, and 13–21 testes from 13–21 fetuses for 10–12 GW. ANOVAs with a random fetus effect were performed using unstructured covariance matrices. A Wilcoxon test was also performed for pairwise comparisons (**p < 0.01, ***p < 0.001). (H) Seminal vesicle weight and (I) plasma testosterone production in host mice carrying human fetal testis xenografts (14–17 GW; n = 4 fetuses) after 7 days (7d) of exposure to vehicle (Corn Oil; open circles) or ibuprofen (10 mg/kg 3 times daily; closed circles) with overall mean ± SEM for vehicle (white bars) and ibuprofen (black bars). Values are means ± SEM from 4 fetuses. Data analyzed by two-way ANOVA.