title: Drug ligand-induced activation of translocator protein (TSPO) stimulates steroid production by aged brown Norway rat Leydig cells.
abstract: Translocator protein (TSPO; 18 kDA) is a high-affinity cholesterol-binding protein that is integrally involved in cholesterol transfer from intracellular stores into mitochondria, the rate-determining step in steroid formation. Previous studies have shown that TSPO drug ligands are able to activate steroid production by MA-10 mouse Leydig tumor cells and by mitochondria isolated from steroidogenic cells. We hypothesized herein that the direct, pharmacological activation of TSPO might induce aged Leydig cells, which are characterized by reduced T production, to produce significantly higher levels of T both in vitro and in vivo. To test this, we first examined the in vitro effects of the TSPO selective and structurally distinct drug ligands N,N-dihexyl-2-(4-fluorophenyl)indole-3-acetamide (FGIN-1-27) and benzodiazepine 4'-chlorodiazepam (Ro5-4864) on steroidogenesis by Leydig cells isolated from aged (21-24 months old) and young adult (3-6 months old) Brown Norway rats. The ligands stimulated Leydig cell T production significantly, and equivalently, in cells of both ages, an effect that was significantly inhibited by the specific TSPO inhibitor 5-androsten-3,17,19-triol (19-Atriol). Additionally, we examined the in vivo effects of administering FGIN-1-27 to young and aged rats. In both cases, serum T levels increased significantly, consistent with the in vitro results. Indeed, serum T levels in aged rats administered FGIN-1-27 were equivalent to T levels in the serum of control young rats. Taken together, these results indicate that although there are reduced amounts of TSPO in aged Leydig cells, its direct activation is able to increase T production. We suggest that this approach might serve as a therapeutic means to increase steroid levels in vivo in cases of primary hypogonadism.
authors.affiliation: Department of Biochemistry and Molecular Biology, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: J Y
authors.initials: JY
authors.lastname: Chung
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.forename: A
authors.initials: A
authors.lastname: Midzak
authors.forename: A L
authors.initials: AL
authors.lastname: Burnett
authors.forename: V
authors.initials: V
authors.lastname: Papadopoulos
authors.forename: B R
authors.initials: BR
authors.lastname: Zirkin
mesh_terms: D000367:Age Factors; D000737:Androstenols / Q000494:pharmacology; D000818:Animals; D001570:Benzodiazepinones / Q000378:metabolism / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D007210:Indoleacetic Acids / Q000378:metabolism / Q000494:pharmacology*; D007985:Leydig Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D051381:Rats; D011914:Rats, Inbred BN; D018079:Receptors, GABA / Q000378:metabolism*; D013256:Steroids / Q000096:biosynthesis*; D013737:Testis / Q000166:cytology
chemical_list: C582440:5-androsten-3,17,19-triol; D000737:Androstenols; D001570:Benzodiazepinones; C463819:Bzrp protein, mouse; D007210:Indoleacetic Acids; D018079:Receptors, GABA; D013256:Steroids; C081306:N,N-di-n-hexyl-2-(4-fluorophenyl)indole-3-acetamide; C028513:4'-chlorodiazepam