title: Mifepristone modulation of ACTH and CRH regulation of bovine adrenocorticosteroidogenesis in vitro.
abstract: Mifepristone (RU486), bovine corticotropin-releasing hormone (CRH), arginine vasopressin (VP), adrenocorticotropin (ACTH1-24), and protein kinase activators (forskolin, [FSK]; phorbol 12-myristate 13-acetate [PMA]) were used in vitro to investigate their direct effect on adrenocorticosteroidogenesis. Bovine adrenocortical fasciculata/reticularis cells (2 x 10(5) viable cells/well) were cultured for 3 d in medium supplemented with 10% fetal calf serum. After incubation for an additional 24 hr in serum-free medium, cells were treated with serum-free medium alone (Control) or various concentrations of ACTH, CRH, VP, FSK, PMA, RU486, and/or various concentrations for 1, 2, 4, or 24 hr. Medium content of cortisol and progesterone were determined by radioimmunoassays. ACTH, CRH, FSK, and PMA each stimulated (P < 0.05) secretion of cortisol in time- and dose-related manners. Although these agents stimulated (P < 0.05) secretion of progesterone in a dose-related manner, medium content of progesterone declined (P < 0.05) over time. The minimal effective doses of ACTH and CRH required to stimulate (P < 0.05) secretion of cortisol relative to the Control over a 4-hr culture period were 0.01 nM and 3 nM, respectively. Relative to observations at 1 hr posttreatment, 24-hr treatment with ACTH or CRH increased the medium content of cortisol by an additional 19.8- and 48-fold, respectively (whereas content of progesterone declined over that time period). VP-stimulated secretion of cortisol was time- (P < 0.05) but not dose-related. Specifically, by 24-hr posttreatment, the medium content of cortisol was increased (P < 0.05) 4.6-fold relative to the quantity of cortisol secreted by 1-hr postaddition of VP (0.01 to 1 microM). Co-treatment with RU486 (1 microM) decreased (p < 0.05) FSK-, ACTH- and CRH-stimulated secretion of cortisol by 77, 27, and 56%, respectively. Similarly, the stimulatory effects of ACTH and CRH on progesterone secretion were reduced (P < 0.05) by 40 and 22%, respectively, by co-addition of RU486. The inhibitory action of RU486 on production of cortisol was no longer apparent by 24 hr after treatment. These observations indicate that RU486 can act as a steroid agonist and as well as an antagonist. These data characterize time- and dose-related direct actions of ACTH, CRH, and RU486 on adrenocorticosteroidogenesis. This information will assist efforts to clarify complex intra-adrenal interactions of neurohormones, growth factors, and endogenous steroids.
authors.affiliation: Department of Animal Science, Texas A&M University System, College Station 77843, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Carroll
authors.forename: S T
authors.initials: ST
authors.lastname: Willard
authors.forename: B L
authors.initials: BL
authors.lastname: Bruner
authors.forename: N H
authors.initials: NH
authors.lastname: McArthur
authors.forename: T H
authors.initials: TH
authors.lastname: Welsh
mesh_terms: D000302:Adrenal Cortex / Q000187:drug effects* / Q000378:metabolism; D000305:Adrenal Cortex Hormones / Q000096:biosynthesis*; D000818:Animals; D001127:Arginine Vasopressin / Q000494:pharmacology; D002417:Cattle / Q000502:physiology*; D005576:Colforsin / Q000494:pharmacology; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology*; D003366:Cosyntropin / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D015735:Mifepristone / Q000494:pharmacology*; D011374:Progesterone / Q000096:biosynthesis; D047428:Protein Kinase Inhibitors; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000305:Adrenal Cortex Hormones; D004791:Enzyme Inhibitors; D047428:Protein Kinase Inhibitors; D001127:Arginine Vasopressin; D003366:Cosyntropin; D005576:Colforsin; D015735:Mifepristone; D011374:Progesterone; D003346:Corticotropin-Releasing Hormone; D013755:Tetradecanoylphorbol Acetate