Molecular adaptations of testosterone-producing Leydig cells during systemic in vivo blockade of the androgen receptor

•Androcur prevented testosterone-reduced Leydig cells steroidogenic capacity/activity.•Androcur completely abolished testosterone-reduced expression of Tspo,StAR,Hsd3b1/2.•Androcur abolished testosterone-reduced expression of Nur77,Gata4,Dax1 transcripts.•Androcur abolished testosterone-induced increase of Hsd3b5/HSD3B and Ar/AR.•Androcur abolished testosterone-reduced cAMP and milieu of cAMP signaling elements. This study systematically evaluates the effects of androgen receptor (AR) blockade on molecular events in Leydig cells. Results showed that intramuscular administration of testosterone-enanthate, at clinically relevant dose, decreased testosterone in interstitial fluid and Leydig cells from adult rats. AR-blocker (Androcur) prevented this effect and testosterone-reduced Leydig cells steroidogenic capacity/activity. Testosterone-reduced expression of some steroidogenic enzymes/proteins (Tspo,StAR,Hsd3b1/2) and transcription factors (Nur77,Gata4,Dax1) was completely abrogated, while decreased expression of Star,Cyp11a1,Cyp17a1,Hsd17b4,Creb1a was partially prevented. In the same cells, increased expression of Hsd3b5/HSD3B and Ar/AR was abolished. Androcur-treatment abolished testosterone-reduced cAMP, coupled with a changed expressional milieu of cAMP signaling elements. Results from in vitro experiments suggest that some of these effects are testosterone-AR dependent, while others could be due to disturbed LH and/or other signals. Presented data provide new molecular insight into Leydig cells function and are important in terms of human reproductive health and the wide-spread use of Androcur as well as use/abuse of testosterone-enanthate.