Androgen regulation of progestin biosynthetic enzymes in FSH-treated rat granulosa cells in vitro.

The influence of androgens on the FSH modulation of progestin biosynthetic enzymes was studied in vitro. Granulosa cells obtained from immature, hypophysectomized, estrogen-treated rats were cultured for 3 days in a serum-free medium containing FSH (20 ng/ml) with or without increasing concentrations (10(-9)-10(-6)M) or 17 beta-hydroxy-5 alpha-androstan-3-one (dihydrotestosterone; DHT), 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha-diol), or the synthetic androgen 17 beta-hydroxy-17-methyl-4,9,11-estratrien-3-one (methyltrienolone; R1881). FSH treatment increased progesterone and 20 alpha-hydroxy-4-pregnen-3-one (20 alpha-OH-P) production by 10.2- and 11-fold, respectively. Concurrent androgen treatment augmented FSH-stimulated progesterone and 20 alpha-OH-P production in a dose-related manner (R1881 greater than 3 alpha-diol greater than DHT). In the presence of an inhibitor of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), the FSH-stimulated pregnenolone (3 beta-hydroxy-5-pregnen-20-one) production (a 20-fold increase) was further enhanced by co-treatment with R1881, 3 alpha-diol or DHT. Furthermore, FSH treatment increased 4.4-fold the activity of 3 beta-HSD, which converts pregnenolone to progesterone. This stimulatory action of FSH was further augmented by concurrent androgen treatment. In contrast, androgen treatment did not affect FSH-stimulated activity of a progesterone breakdown enzyme, 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD). These results demonstrate that the augmenting effect of androgens upon FSH-stimulated progesterone biosynthesis is not due to changes in the conversion of progesterone to 20 alpha-OH-P, but involves an enhancing action upon 3 beta-HSD/delta 5,delta 4-isomerase complexes and additional enzymes prior to pregnenolone biosynthesis.