Dehydroepiandrosterone Sulfate Production Research Articles

Functional zonation of the midgestation human fetal adrenal cortex: Fetal versus definitive zone use of progesterone for cortisol synthesis

Studies of human fetal adrenal function and its control have revolved mainly around the remarkable capacity of the unique fetal zone of this gland to elaborate dehydroepiandrosterone sulfate. Another important function of the fetal adrenal, however, is its production of cortisol. Because the human fetal adrenal is deficient in 3β-hydroxysteroid dehydrogenase activity, cortisol has been thought to be formed from circulating progesterone. To further investigate this hypothesis, cortisol production by separated fetal and definitive zones of the midgestation human fetal adrenal in organ culture has been examined in the absence and presence of varying concentrations of progesterone and adrenocorticotropic hormone. Cortisol was measured by radioimmunoassay. In the absence of progesterone, cortisol production by both zones increased gradually over time in culture in response to adrenocorticotropic hormone. In the presence of progesterone, cortisol production by the definitive zone was unchanged. In contrast, the response of the fetal zone to progesterone was immediate: cortisol production increased significantly and remained high throughout the culture period. These results suggest a greater capacity of the fetal zone to utilize progesterone for cortisol production and are consistent with morphologic evidence that the active zone of the midgestation human fetal adrenal is the fetal zone, possessing not only the enzyme activity necessary for dehydroepiandrosterone sulfate production but, except for 3β-hydroxysteroid dehydrogenase, that for cortisol production as well.

Androgen production in primary culture of immature porcine leydig cells

The present paper examines the steroidogenic responsiveness of immature porcine Leydig cells in primary culture. Both testosterone (T) and dehydroepiandrosterone sulfate (DHAS) secretion were measured under basal conditions and after stimulation with human chorionic gonadotropin (hCG) (25 ng/ml). In medium supplemented with insulin, transferrin, epidermal growth factor (3H) and 0.1% calf serum, cells survived 3–5 days in culture. The production of steroids (under hCG stimulation) is poor on day 0–1 of the culture. On day 2–4 basal T and DHAS levels are 1.9 and 17.0 ng/10 6 cells/24 h. The addition of hCG stimulated T and DHAS production 19- and 6-fold respectively and the average productions were 37 and 109 ng/10 6 cells/24 h. Increasing the serum to 0.5% did not change the viability of the cultures, but increased hCG stimulated T and DHAS production (183 and 188 ng/10 6 cells/24 h). The addition of α-tocopherol (vitamin E) to 0.1% calf serum led to a 4-fold increase in stimulated T production (142 ng/10 6 cells/24 h) and maintained full cell viability for more than 5 days. Measurement of 3β-ol steroid dehydrogenase activity indicates that the amount of enzyme is 4 times higher at day 2 than at day 0 and 1 (with or without hCG), suggesting a spontaneous maturation of the cells in culture. This might explain the increased T production with time in culture. In cumulative experiments (24 h) the cells do not seem to be desensitized to hCG stimulation following prolonged exposure to 25 ng hCG since the daily steroid production is increasing with time in culture. However, kinetic studies show that steroidogenesis is not linear over a 24 h period. In cumulative experiments the steroid production stops between 12 and 16 h following hCG exposure (5 and 100 ng/ml) and resumes following a medium change. These results suggest that some inhibitory compounds are accumulated in the medium and are controlling the Leydig cell function. Moreover high doses of hCG (100 ng/ml) result in a lower production of steroids and an earlier plateau in the case of DHAS. These results demonstrate that porcine Leydig cells can live and differentiate in hormone- and vitamin-supplemented medium and that auto-feedback mechanisms inhibiting steroid accumulation take place under in vitro conditions.

Spermatic and peripheral venous plasma concentrations of dehydroepiandrosterone sulfate in prepubertal and pubertal boys

Dehydroepiandrosterone sulfate (DEAS), a major adrenal product, is quantitatively one of the most important steroids found in human testicular tissue. However, conflicting data have been reported about the testicular production of DEAS ‘ in vivo’. We have measured the spermatic and peripheral concentrations of testosterone (T) and DEAS in two groups of prepubertal (Group I, N = 18) and pubertal (Group II; N = 11) boys undergoing surgery for undescended testis, inguinal hernia or varicocele. Mean (± SE) spermatic concentrations of DEAS (77 ± 16 and 113 ± 19 μg/dl in Group I and II respectively) were not significantly different from peripheral concentrations (70 ± 13 and 130 ± 16 μg/dl in Group I and II respectively). Mean spermatic concentrations of T (153 ± 101 and 7515 ± 4314 ng/dl in Group I and II respectively) were significantly different from peripheral concentrations (9 ± 1 and 149 ± 53 ng/dl; P < 0.001 in both groups). Spermatic and peripheral levels of T and DEAS found in prepubertal boys were significantly lower than those found in pubertal boys. Spermatic levels of DEAS were not significantly related with spermatic levels of T in both groups. Our data show that, as in adult subjects, no significant spermatic-peripheral DEAS gradient is present in prepubertal and pubertal boys.

Umbilical cord plasma levels of dehydroepiandrosterone sulfate during human gestation.

In the current investigation we sought to evaluate fetal adrenal dehydroepiandrosterone sulfate (DS) production during normal human pregnancy. We found that the mean umbilical cord plasma concentration of DS was reasonably constant from 18-34 weeks of gestation, ranging from 1131-1517 ng/ml. Thereafter, the cord plasma DS concentration rose steadily to 2463 +/- 665 ng/ml (mean +/- SD) at 39-40 weeks of gestation. Between 20 and 32 weeks of gestation, the estimated fetal plasma DS pool increased from 8 to 146 microgram. At term, the estimated fetal DS plasma pool was in excess of 300 microgram. From 34-40 weeks of gestation, the estimated fetal DS pool size and the fetal plasma DS concentration increased coincidentally with a marked rise in the known rate of fetal adrenal growth. From these findings, we conclude that fetal adrenal DS production increases steadily throughout gestation, and the rate of increase is probably dependent upon the rate of fetal adrenal growth. Furthermore, the rapid increase in maternal estrogen levels near term observed by numerous investigators can be explained in part by an increased availability of the fetal estrogen precursor DS at this time.

In utero evidence for a functional fetoplacental unit in rhesus monkeys.

To test the hypothesis that a fetoplacental unit for estrogen biosynthesis is operative in the rhesus monkey, studies were performed in chronically catheterized animals during late gestation. Changes in fetal and maternal plasma concentrations of estrone, estradiol, dehydroepiandrosterone sulfate (DHEAS), and androstenedione were determined in response to infusions of dexamethasone into mothers, and of ACTH, DHEAS, or androstenedione into fetuses. A significant (P<0.0l) maternal: fetal concentration difference (∿20:1) was observed in control samples for estradiol, but not for estrone, DHEAS, or androstenedione (P>0.05). Dexamethasone administration decreased (P<0.05) maternal estrone, estradiol, and DHEAS, as well as fetal estrone and DHEAS. Subsequent administration of ACTH to fetuses increased (P<0.05) fetal estrone, DHEAS and androstenedione, and also maternal estrone and estradiol. Fetal estradiol and maternal DHEAS and androstenedione were not significantly changed (P>0.05). The concentrations of fetal estrone and DHEAS, and of maternal estrone and estradiol returned to pre-dexamethasone control values during infusion of ACTH into fetuses. Administration of DHEAS or androstenedione to fetuses without concurrent dexamethasone administration increased (P<0.05) fetal estrone and maternal estrone and estradiol. No significant changes (P>0.05) were observed in maternal or fetal cortisol or progesterone concentrations. Fetal estradiol levels were relatively low in all of the experiments, and they were not significantly altered by dexamethasone, ACTH, DHEAS, or androstenedione infusions (P>0.05). Collectively, these data indicate that the increases in fetal and maternal estrogens after administration of ACTH to fetuses are the result of ACTH stimulation of fetal adrenal DHEAS and androstenedione production. Androstenedione and DHEAS are transported via the fetal circulation to the placenta, where they are converted to estrogens. Thus, the rhesus monkey, like the human, has a functional fetoplacental unit.

Placental origin of the suppression of 3β-hydroxysteroid dehydrogenase in the fetal zone cells of human fetal adrenals

The present study investigates the regulation of human fetal adrenal steroidogenesis. Adrenals were cultured alone or together with placental tissue. The main secretory products of adrenals were analysed by gas liquid chromatography from tissue culture medium. ACTH stimulated both the cortisol and dehydroepiandrosterone sulphate (DHAS) production of cultured adrenals, but the presence of placental tissue in the adrenal cultures inhibited effectively cortisol production ( P < 0.001) and increased the production of free androgens (dehydroepiandrosterone, androstenedione). Human chorionic gonadotropin (hCG) was not responsible for the inhibitory effect of the placenta on cortisol production. It can be concluded that ACTH is the stimulus of human adrenal at fetal age, but a placental factor inhibits the 3β-hydroxysteroid dehydrogenase Δ5-Δ4 isomerase system (3β-HSD) in the fetal zone and directs steroidogenesis in vivo towards dehydroepiandrosterone (DHA) and DHAS production.

The effects of progesterone, pregnenolone, estriol, ACTH and hCG on steroid secretion of cultured human fetal adrenals

Endogenous steroid secretion and the conversion of exogenous pregnenolone and progesterone were studied in tissue culture of human mid-term fetal adrenals. Addition of pregnenolone on the first day of cultivation caused an increase in DHAS (dehydroepiandrosterone sulphate) and pregnenolone sulphate secretion during the first cultivation days, but no significant increase in cortisol production was noted. Progesterone at the same stage of cultivation was effectively converted into cortisol indicating that human fetal adrenals in spite of the lack of 3β-HSD (3β-OH-steroid dehydrogenase) are capable of synthesizing efficiently cortisol using exogenous ( in vivo placental) progesterone as a substrate. During later stages of cultivation pregnenolone was converted into pregnenolone sulphate, but not into DHAS, indicating that sulphokinase activity is maintained in cultured adrenal cells. Estriol and estradiol-17β inhibited steroidogenesis in ACTH-stimulated tissue culture, and the inhibition step seemed to be 3β-HSD-step. The demanded concentrations of estrogens for significant inhibition were, however, unphysiological. Estriol-3-sulphate and DHAS did not inhibit steroidogenesis. hCG did not stimulate DHA or DHAS production, neither did it modify the stimulatory effect of ACTH towards these estrogen precursors.

Pattern of Plasma Dehydroepiandrosterone Sulfate Levels in Humans from Birth to Adulthood: Evidence for Testicular Production*

Plasma levels of dehydroepiandrosterone sulfate (DHAS) were measured in 513 normal full term newborns, infants, children, adolescents, and adults and the results were expressed in micrograms per dl. In infancy and childhood, DHAS levels were similar in both sexes. In 74 neonates, mixed cord blood mean values /+- SD were 134.6 +/- 64. During the first day of life, plasma DHAS levels were 140 +/- 125 in 33 neonates. During the first month of life, DHAS decreased drastically, then more progressively until the 6th month of life. Between 1-6 months of age, mean levels were 5.9 +/- 4.7 in 40 children. DHAS was very low between 1-6 yr of life (2.3 +/- 1.6) and rose abruptly at the 7th year of life. Thereafter, DHAS continued to increase correlatively with age and pubertal stages in both sexes, a further increase after age 16 or pubertal stage P5 was noted only in male subjects. In adults, DHAS was significantly higher in male (224 +/- 93) than in female (138.3 +/- 51) subjects. DHAS levels were compared to those of dehydroepiandrosterone; at two periods of life, early infancy and adulthood, their patterns differed. After long term hCG stimulation, DHAS increased significantly in 45 normal prepubertal boys and in 2 boys with adrenal insufficiency. These data would suggest a direct testicular production of DHAS.

Role of hCG in Regulation of the Fetal Zone of the Human Fetal Adrenal Gland*

It has been suggested that hCG is a trophic hormone for the fetal zone of the human fetal adrenal gland. To test this hypothesis, the isolated fetal zones of adrenals from eight fetuses (12-17-week gestation age) were superfused in the presence or absence of hCG. Dehydroepiandrosterone sulfate (DHAS) was measured in the superfusion effluent. A significant increase in DHAS production was observed in the presence of hCG. DHAS secretion decreased during the first 60 min in the control and experimental superfusions from 83 +/- 10.0 (mean +/- SE) to 71 +/- 8.0, and from 90 +/- 9.0 to 70 +/- 6.0 ng/100 mg/ml, respectively. In the presence of hCG (250 ng/ml), DHAS secretion increased significantly (P less than 0.01) over the controls to 116 +/- 12.0 at 120 min, and remained above the controls thereafter. These results support the hypothesis that hCG is one of the regulators of DHAS production by the human fetal adrenal gland early in gestation. As we found that ACTh stimulated DHAS secretion in a previous study and as there is indirect evidence for a role of ACTH in DHAS regulation late in pregnancy, these observations suggest dual regulation by hCG and ACTH early in pregnancy, and a possible transition to ACTH regulation of the fetal zone of the human fetal adrenal after midgestation.

Exploration of the human fetal pituitary adrenal axis: Stimulation of cortisol and dehydroepiandrosterone sulfate biosynthesis by homologous pituitary in organ culture

Adrenals obtained from human abortices at midpregnancy were kept under conditions of tissue culture and the production of cortisol and dehydroepiandrosterone sulfate (3β-hydroxy-5-androsten-17-one, 3-sulfate; DHA-S) monitored by radioimmunoassays for up to 2 weeks. Basal production of dehydroepiandrosterone sulfate was considerably higher than that of cortisol. α 1−24corticotrophin, α 1−39corticotrophin, (a long acting porcine corticotrophin) at the concentration of 1 mU/ml of culture medium in both instances, and dibutyryl cyclic AMP (1mM)enhanced the production of both steroids some 3 to 30 times above control values. Medium harvested from homologous pituitary cultures had comparable corticotrophic activity. It is concluded that at midpregnancy the regulation of corticoidogenesis implies the existence of a corticotrophic factor either identical or closely related to ACTH.

Plasma androgens in women with hyperprolactinaemic amenorrhoea.

Cortisol, androstenedione, testosterone, dehydroepiandrosterone sulphate (DHAS) and free dehydroepiandrosterone (DHA) were measured in plasma of ten women affected by amenorrhoea with hyperprolactinaemia and eleven women affected by secondary hypothalamic amenorrhoea; twelve normal women at the second day of the menstrual cycle were used as controls. All subjects were hospitalized and 17-ketosteroids, 17OH-corticosteroids and total dehydroepiandrosterone were also measured in urine. Plasma DHAS was increased in all subjects affected by amenorrhoea with hyperprolactinaemia, while plasma DHA and urinary DHA were significantly increased in this group in comparison to other groups. Plasma cortisol, androstenedione and testosterone and urinary 17-oxosteroids and 17OH-corticosteroids were not significantly differnt in the three groups. In subjects affected by amenorrhoea with hyperprolactinaemia treated with bromocriptine a clear decrease of DHAS correlating with a decrease of plasma prolactin was observed. Since in wome DHAS sems to be almost exclusively secreted by the adrenal gland and most of the circulating DHA is dervied from adrenal secretion, these data suggest that human prolactin can stimulate DHAS production by the adrenal cortex.

The contribution of plasma dehydroepiandrosterone sulfate to testosterone in a virilized patient with an arrhenoblastoma.

ABSTRACT The conversion of dehydroepiandrosterone sulfate (DS) to testosterone (T) and the DS production rate were quantified preand postoperatively in a virilized 32-yr-old female with an arrhenoblastoma. Additionally, at the time of operation, the tumor was isolated and perfused with DS-4-14C and T-7α-3H and the venous effluent collected. Preoperatively, the maximum quantity of plasma T derived from circulating DS was 640 μg/day. Postoperatively, this figure was found to be 310 7mu;g/day. The preoperative DS production rate was 80 mg/day, with the postoperative production rate falling to 15.5 mg/day. Labeled DS perfused through the tumor was not converted to T, testosterone sulfate or androstenedione. A small conversion of DS to dehydroepiandrosterone was noted. It is concluded that the conversion of circulating DS to T by the tumor itself was not an important factor in the genesis of this patient's virilism. Likewise, the peripheral conversion of circulating DS to T did not appear to be significant in ...

13 Plasma 17-Ketosteroid Levels During Adolescence

To study adrenal maturation at adolescence, pooled blood was collected from prepubertal males, age 6–9 years, and from pubertal boys, age 12–14 years. Plasma steroid sulfates were extracted in the form of methyl green salts, which were solvolyzed and then successively subjected to purification by means of the Girard T-reagent, digitonin, thin-layer chromatography, and gas-liquid chromatography (free steroid, acetate, trimethylsilyl ether) on the phases SE-52 and QF-1. In the 6–9 year-old pool the level of androsterone sulfate (AS), corrected for 80 % recovery, was 8.6 μg%; and of dehydroepiandrosterone sulfate (DHAS), corrected for 65% recovery, 13.5 μg%. In the 12–14 year-old male pool the level of AS was 56.7 μg % and of DHAS 80.3 μg %. In adult males, studied for comparison, plasma levels of AS were 40–53 μg % and of DHAS 89–209 μg%. No etiocholanolone sulfate or free 17-ketosteroid was detected.The dramatic rise of plasma DHAS and AS during adolescence appears to occur simultaneously wih beginning testicular maturation. Since ACTH secretion is not known to change at the time of adolescence, these observations suggest that beginning adrenal production of DHAS at this age is not directly related to ACTH stimulation, but rather that it is promoted by some other trophic factor. (SPR)

ÉTUDES IN VIVO SUR LE MÉTABOLISME DES ANDROGÉNES APRES ADMINISTRATION DE STÉROÏDES INHIBITEURS DE L'OVULATION

ABSTRACT Normal subjects and hirsute women with micropolycystic ovaries were treated with ethinyl-oestrenol + 3-methoxy-ethinyl-oestradiol (Lyndiol®), in view of studying the action of this compound on the production of androgens and on the urinary excretion of their metabolites. In normal men, the production of testosterone and the excretion of androsterone and aetiocholanolone are suppressed, whereas the excretion of other 17-ketosteroids and the production of dehydroepiandrosterone sulphate are unchanged. Moreover, the luteinizing hormone activity (LH) in plasma is depressed. It seems that the preparation inhibits specifically the testicular androgen production, by suppressing the hypothalamo-hypophyseal control of LH. Testosterone production and urinary 17-ketosteroid excretion are modified in the same way in women with Stein-Leventhal's syndrome. Physiopathological and therapeutical implications which come from these results are discussed.