Fig. 8
Long-term effect of CBD and/or THC on Leydig cell biology. (A) Representative immunostaining of AMH, LIN28A, and CYP11A1 in 10–12 DW testis in dark brown. Sections were counterstained with hematoxylin (in blue). Scale bar = 100 µm. Quantification of the surface area occupied by (B) cleaved caspase-3-positive cells (Cas3 +) or (C) CYP11A1 + cells after Control (Ctrl), CBD, or THC treatment at 10−5 M (CBD, THC) or a mixture of CBD and THC at 10−5 M (C/T5) or 10−6 M (C/T6) compared to respective control. (D) Density of LIN28-positive cells (LIN28 +), resulting from the quantification of positive cells per surface of intracordonal tissue after CBD and/or THC treatment. (E) Quantification of the surface area occupied by AMH-positive cells (AMH +) compared to section area after CBD and/or THC treatment. (F) Amount of testosterone in the supernatants of 10–12 DW fetal human testis explants cultured for 4 (D4), 8 (D8), and 14 (D14) days or (G) for 8 days for AMH in presence of solvent control (DMSO and EtOH) or 10−5 M of CDB, THC, or a mixture of CBD/THC (ratio 1:1) at the concentrations of 10−6 and 10−5 M. Results are expressed related to the amount of testosterone or AMH produced during the first day of the culture (D0). Data are means + / − SEM. An ANOVA test was performed followed by a Freidman test for pair comparisons (*, p < 0.05; **, p < 0.01;***, p < 0.001; ****, p < 0.0001)