Fig. 5
Effect of CBD and/or THC on Leydig cell functions. Amount of testosterone in the supernatants of fetal human testis explants of 8–10 (gray line) or 10–12 (black line) DW cultured for 72 h in presence of controls (DMSO and/or EtOH) or 10−7 to 10−5 M of (A) CDB, (B) THC, or (C) a mixture of CBD/THC (ratio 1:1). Results are expressed related to the amount of testosterone produced during the first 24 h of culture (D0) and as fold from the respective control. Data are means + / − SEM. An ANOVA test was performed followed by Friedman test for pair comparisons (*, p < 0.05; ***, p < 0.01; **p < 0.001; ****, p < 0.0001). (D) Concentration of insulin-like factor 3 (INSL3) in the supernatants of fetal human testis explants of 8–10 or 10–12 DW cultured for 72 h in presence of controls or 10−5 M of CBD, THC, or a mixture of CBD/THC (ratio 1:1). Results are expressed related to the amount of INSL3 produced during the first 24 h of culture (D0) and as fold from the respective controls. Data are means + / − SEM. Control and treated conditions were compared two by two using Wilcoxon tests. (*, p < 0.05). (E) Representative immunostaining of CYP11A1 in 8–10 and a 10–12 DW human fetal testis in controls (DMSO/EtOH) and CBD/THC (ratio 1:1) conditions. CYP11A + cells appear dark brown (DAB staining) in extracordonal tissue, and sections were counterstained with hematoxylin (in blue). No change in terms of Leydig cell morphology was observed. Scale bar = 100 µm. (F) Quantification of the surface area occupied by CYP11A1 + Leydig cells related to the section surface after 10−5 M of CBD and/or THC treatment compared to respective controls in 8–10 and 10–12 DW embryonic testis