Dietary resistant maltodextrin ameliorates testicular function and spermatogenesis in streptozotocin-nicotinamide-induced diabetic rats.

This study investigated the effect of resistant maltodextrin (RMD) on reproduction in streptozotocin (STZ)-nicotinamide-induced type 2 diabetic male rats. Forty male rats were induced with diabetes by a single intraperitoneal injection of STZ (50 mg kg(-1)) and nicotinamide (100 mg kg(-1)). Five groups were analysed in total: normal, diabetic rats without RMD, diabetic rats with RMD 1.2 g per 100 g diet (1×), with RMD 2.4 g per 100 g (2×), and with RMD 6.0 g per 100 g (5×). The groups of diabetic rats with the RMD supplement, compared to those without supplement, showed improved plasma glucose control, attenuated insulin resistance and recovery of testosterone level and spermatogenesis stage. The STZ-nicotinamide-induced diabetes mellitus (DM) caused a significant reduction in serum testosterone, testis androgen receptor (AR), steroidogenic acute regulatory protein (StAR) and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) protein, but a statistical recovery in each of these was observed in the 5× group. TUNEL-positive cells were observed in the diabetic without RMD group, and RMD treatment reduced apoptotic germ cells. The expression of Bax/Bcl2 was induced in the diabetic group and also significantly reduced in the 5× group. Dietary RMD may improve metabolic control in STZ-nicotinamide-induced diabetic rats and attenuate hyperglycaemia-related impaired male reproduction and testicular function.

Protective role of Nigella sativa oil against reproductive toxicity, hormonal alterations, and oxidative damage induced by chlorpyrifos in male rats.

This study is aimed at elucidating the possible protective effects of Nigella sativa oil (NSO) in alleviating the toxicity of chlorpyrifos (CPF) on reproductive performance in male rats. Animals were orally administered with NSO (1 ml/kg/day), CPF (20 mg/kg/day), and NSO + CPF every day for 4 weeks. Results showed that CPF decreased spermatid number, sperm count, daily sperm production, and sperm motility while increased dead sperm and abnormal sperm compared with the control. Also the levels of testosterone, thyroxine levels, steroidogenic enzyme 17-ketosteroid reductase, body weight, food intake, and relative weight of reproductive organs were decreased. Thiobarbituric acid reactive substances were increased, while glutathione (GSH) and antioxidant enzymes were decreased in plasma and testes of rats treated with CPF. Histopathological examination of testes showed a decrease in the number of seminiferous tubules, form shrinkage, enlargement of the connective tissue and gametogenic changes in germ cells of rats treated with CPF. NSO alone increased testosterone, semen characteristics, GSH, and antioxidant enzymes and decreased the levels of free radicals. Furthermore, the presence of NSO with CPF alleviates its toxic effects. Our results indicated that NSO can improve semen picture and moderate CPF-induced reproductive toxicity.

Protective role of L-carnitine and vitamin E on the testis of atherosclerotic rats.

Atherosclerosis is a condition caused by lipid build-up and inflammation in the arteries, so hyperlipidemia is the major reason for atherosclerosis. Testis was found to be negatively affected by hyperlipidemia which leads to its impaired functions. Vitamin E and l-carnitine have well-known lipid-lowering and antioxidative activities. Triton WR 1339 is a non-ionic detergent, which induces severe hyperlipidemia by inhibition of lipoprotein lipase. The present study evaluates the protective role of vitamin E and l-carnitine on the testis in atherosclerosis and detects the most effective choice for protection against atherosclerosis; vitamin E, l-carnitine or a combination of both. A total of 80 albino male rats were divided into eight groups (10 rats for each group): control (G1), triton (G2), l-carnitine (G3), triton + l-carnitine (G4), vitamin E (G5), triton + vitamin E (G6), l-carnitine + vitamin E (G7) and triton + l-carnitine + vitamin E (G8). Data showed a significant increase in the levels of total cholesterol (TC), triglycerides (TGs), low-density lipoprotein cholesterol (LDL-C), 17 beta hydroxysteroid dehydrogenase (17 ß HSD), testicular catalase and malondialdehyde (MDA) in G2 when compared with G1, whereas high-density lipoprotein cholesterol (HDL-C), serum testosterone, testicular 17 ketosteroid reductase (17 KSR), total thiol and glutathione-S-transferase (GST) data showed a significant decrease in G2 when compared with G1. Treatment with l-carnitine or/and vitamin E helps in improving the adverse effect of triton; also the histological changes confirm this finding. So the present study recommends all people to include l-carnitine and vitamin E in their diet to be protected against atherosclerosis.

A novel nonsense mutation in HSD17B3 gene in a Tunisian patient with sexual ambiguity.

INTRODUCTION: 17ß-hydroxysteroid dehydrogenase type 3 (HSD17B3) isoenzyme is present almost exclusively in the testes and converts delta 4 androstenedione to testosterone. Mutations in the HSD17B3 gene cause HSD17B3 deficiency and result in 46,XY Disorders of Sex Development (46,XY DSD). AIM: This study aimed to present the clinical and biochemical features of a Tunisian patient who presented a sexual ambiguity orienting to HSD17B3 deficiency and to search for a mutation in the HSD17B3 gene by DNA sequencing. METHODS: Polymerase chain reaction (PCR) amplification and subsequent sequencing of all the coding exons of HSD17B3 gene were performed on genomic DNA from the patient, her family, and 50 controls. RESULTS: Genetic mutation analysis of the HSD17B3 gene revealed the presence of a novel homozygous nonsense mutation in the exon 9 (c.618 C>A) leading to the substitution p.C206X. The mutation p.C206X in the coding exons supports the hypothesis of HSD17B3 deficiency in our patient. CONCLUSION: The patient described in this study represented a new case of a rare form of 46,XY DSD, associated to a novel gene mutation of HSD17B3 gene. The screening of this mutation is useful for confirming the diagnosis of HSD17B3 deficiency and for prenatal diagnosis.

Polymorphisms in steroidogenesis genes, sex steroid levels, and high myopia in the Taiwanese population.

PURPOSE: To evaluate the relationship among single nucleotide polymorphisms (SNPs) in steroidogenesis enzyme genes, serum levels of sex steroids, and high myopia in Taiwanese male and female populations. METHODS: A campus-based sample of 283 cases (145 males and 138 females) with high myopia and 280 controls (144 males and 136 females) with low myopia or emmetropia was studied. Estradiol, progesterone, and testosterone levels were determined using enzyme-linked immunosorbent assay kits. We genotyped six SNPs within five steroidogenesis enzyme genes (17 alpha-hydroxylase/17,20 lyase [CYP17A1], 3 beta-hydroxysteroid dehydrogenase [HSD3B1], 17 beta-hydroxysteroid dehydrogenase 1 [HSD17B1], steroid-5-alpha-reductase, alpha polypeptide 2 [SRD5A2], and aromatase [CYP19A1]) using polymerase chain reaction-restriction fragment length polymorphism methods. Student's t-tests, χ(2) tests, logistic regression, multifactor dimensionality reduction (MDR) methods, and ANOVA were used to determine significance. RESULTS: An MDR analysis corroborated the synergistic genotype association and demonstrated that synergistic interaction between rs6203 (HSD3B1), rs10046 (CYP19A1), and sex might confer susceptibility to high myopia (p=0.019). In both male and female subjects, levels of testosterone were significantly higher in cases than in controls; in male subjects, the levels of estradiol were significantly higher and those of progesterone were significantly lower in cases (all p-values <0.001). The rs605059 (HSD17B1), with sex-gene interaction, showed association with estradiol levels in males (p=0.035) and testosterone levels in females (p=0.027). CONCLUSIONS: Testosterone levels correlate with high myopia, and interaction of steroidogenesis enzyme genes and sex may be a modulating factor in sex hormone metabolism and high-myopia risk.

Premature Adrenarche in Girls Characterized By Enhanced 17,20-Lyase and 17ß-Hydroxysteroid Dehydrogenase Activities.

CONTEXT: Girls with premature adrenarche (PA) may have a higher risk of developing polycystic ovary syndrome (PCOS) and metabolic syndrome. The biological purpose of adrenarche is unknown and the role of novel biosynthetic pathways remains unclear. OBJECTIVE: To compare the urinary steroid metabolome and enzyme activities of girls with PA to age-matched control girls and to published steroid values of girls with normal adrenarche and of women with PCOS and their newborn daughters. DESIGN: Prospective observational study from 2009 to 2014. SETTING: Academic pediatric endocrinology referral center. PARTICIPANTS: Twenty-three girls with PA and 22 healthy, age-matched girls. MAIN OUTCOME MEASURES: Steroid metabolites in 24-hour urine samples, including 4 progesterones, 5 corticosterones, aldosterone, 13 androgens, 2 estrogens, 14 glucocorticoids, and enzyme activities represented by metabolite ratios. RESULTS: Girls with PA had a higher body mass index (mean standard deviation scores 0.9 vs -0.3, P = 0.013). Androgen excretion was higher in PA girls than in control girls (median 3257 nmol/24 hours vs 1627 nmol/24 hours, P < 0.001), in particular metabolites from alternate androgen pathways. The amount of progesterone, corticosterone, aldosterone, estrogen, and cortisol metabolites were similar between groups. Activities of 17ß-hydroxysteroid-dehydrogenase and of 17,20-lyase were higher in girls with PA. Activities of 3ß-hydroxysteroid-dehydrogenase, 21-hydroxylase, and 5α-reductase activity were not different between groups, in contrast to published results on girls with normal adrenarche or PCOS females. CONCLUSIONS: Metabolites and enzymes involved in alternate androgen pathways appear to be markers of PA. Prospective studies should assess whether steroid production in PA also differs from adrenarche at normal timing and persists into adulthood.

Changes in Adrenal Androgens and Steroidogenic Enzyme Activities From Ages 2, 4, to 6 Years: A Prospective Cohort Study.

CONTEXT: The levels of adrenal androgens are increased through the action of steroidogenic enzymes with morphological changes in the adrenal zona reticularis. OBJECTIVE: We investigated longitudinal changes in androgen levels and steroidogenic enzyme activities during early childhood. DESIGN AND PARTICIPANTS: From a prospective children's cohort, the Environment and Development of Children cohort, 114 boys and 86 girls with available blood samples from ages 2, 4, and 6 years were included. OUTCOME MEASUREMENTS: Serum concentrations of adrenal androgens using liquid chromatography-tandem mass spectrometry and steroidogenic enzyme activity calculated by the precursor/product ratio. RESULTS: During ages 2 to 4 years, 17,20-lyase and dehydroepiandrosterone (DHEA) sulfotransferase activities increased (P < 0.01 for both in boys). During ages 4 to 6 years, 17,20-lyase activity persistently increased, but 3ß-hydroxysteroid dehydrogenase (HSD) and 17ß-HSD activities decreased (P < 0.01 for all). Serum DHEA sulfate (DHEA-S) levels persistently increased from 2, 4, to 6 years, and DHEA, 17-hydroxyprogesterone, and androstenedione levels increased during ages 4 to 6 years (P < 0.01 for all). Serum DHEA-S levels during early childhood were associated with body mass index z-scores (P = 0.001 in only boys). CONCLUSION: This study supports in vivo human evidence of increased 17,20-lyase and DHEA sulfotransferase activities and decreased 3ß-HSD activity during early childhood.

Tissue-specific transcription profiles of sex steroid biosynthesis enzymes and the androgen receptor.

17beta-hydroxysteroid dehydrogenase (17beta-HSD) and 5alpha-reductase isoenzymes play a crucial role in the formation and metabolism of sex steroids. Not only the key androgens testosterone and dihydrotestosterone but also their precursors are potent activators of the androgen receptor and are, therefore, likely to act as determinants of male sexual differentiation and maturation in a differentially regulated way. The aim of the present study was to relatively quantify the expression of the mRNA of 17beta-HSD isoenzymes, namely, type 1, 2, 3, 4, 5, 7, and 10, together with the 5alpha-reductase type 1 and 2, and the androgen receptor in normal human males and females. RNA was isolated from peripheral blood cells of both sexes and from genital skin fibroblasts (GSFs) of two different localizations (foreskin and scrotal skin) obtained from phenotypically normal males. mRNA expression was semi-quantified by quantitative reverse-transcriptase polymerase chain reaction with the LightCycler Instrument (Roche). The examined enzymes show statistically significant differences in their transcription pattern between the blood and the GSF RNA samples. Within the GSF samples, there are also significant variations between the two examined localizations in the transcription of 17beta-HSD type 1, 2, 4, and 5 as well as for the androgen receptor. We found large interindividual variation of enzyme transcription patterns in all investigated tissues. In peripheral blood cells, no sex-specific differences were seen. We conclude that sex steroid enzymes are expressed not only in genital primary target tissues but also in peripheral blood. The expression in different target tissues may contribute to both the individual sexual and tissue-specific phenotype in humans.

Sterilization of male stray dogs with a single intratesticular injection of calcium chloride: a dose-dependent study.

OBJECTIVE: To study a method of chemical sterilization and its efficacy in adult male stray dogs. METHODS: Sterilization was performed 45 days after a single bilateral intratesticular injection of calcium chloride (CaCl(2)) at the doses of 5, 10, 15 or 20 mg per testis per kg body weight. RESULTS: Histomorphological measures of testes showed total necrosis of testicular tissue at 45 days after an injection of either 10 or 15 or 20 mg CaCl(2) along with fibrosis and hyalinization in seminiferous tubules and interstitial spaces. Infiltration of leucocytes was also observed with the 10- or 15-mg dose. Disintegration of germ cell arrangement in seminiferous tubules and washing out of germ cells from the tubules were noted with the 5-mg dose. Relative organ weight, epididymal sperm count, plasma and intratesticular concentrations of testosterone, testicular activities of Delta(5),3beta-hydroxysteroid dehydrogenase (Delta(5),3beta-HSD), 17beta-hydroxysteroid dehydrogenase (17beta-HSD), glutathione peroxidase (GPx), glutathione S-transferase (GST) and superoxide dismutase (SOD) and testicular contents of glutathione (GSH) and glutathione disulphide (GSSG) and the ratio of GSH/GSSG, all were declined in each of the calcium chloride treated groups in comparison to the control group. Increases occurred in testicular malondialdehyde (MDA) and plasma concentrations of LH and FSH with each of the treatments by comparison with the control group. Plasma concentrations of cortisol, fasting blood sugar level, blood urea nitrogen as well as packed cell volume (PCV) and total plasma protein were recorded to monitor the changes in chronic stress in the experimental animals. Changes in these parameters were not significant. CONCLUSION: An intratesticular injection of CaCl(2) at specified doses could be a suitable method of sterilization in preference to surgical castration of dogs.

Loss of PI3K p110α in the Adipose Tissue Results in Infertility and Delayed Puberty Onset in Male Mice.

Deletion of PI3K catalytic subunit p110α in adipose tissue (aP2-Cre/p110αflx/flx, α-/- hereafter) results in increased adiposity, glucose intolerance, and liver steatosis. Because this endocrine organ releases hormones like leptin, which are important in reproductive physiology, we investigated the reproductive phenotype of α-/- males. Compared to controls, α-/- males displayed delayed onset of puberty accompanied by a reduction in plasma LH levels and testicular weight. At postnatal day 30, α-/- mice exhibited normal body weight but elevated fasted plasma leptin levels. Testicular leptin gene expression was increased, whereas expression of the cholesterol transporter StAR and of P450 cholesterol side chain cleavage enzyme was decreased. Adult α-/- males were infertile and exhibited hyperandrogenemia with normal basal LH, FSH, and estradiol levels. However, neither sperm counts nor sperm motility was different between genotypes. The mRNA levels of leptin and of 17-beta-dehydrogenase 3, and enzyme important for testosterone production, were significantly higher in the testis of adult α-/- males. The mRNA levels of ERα, an important regulator of intratesticular steroidogenesis, were lower in the testis of adult and peripubertal α-/- males. We propose that chronic hyperleptinemia contributes to the negative impact that disrupting PI3K signaling in adipocytes has on puberty onset, steroidogenesis, and fertility in males.

Human chorionic gonadotropin stimulation gives evidence of differences in testicular steroidogenesis in Klinefelter syndrome, as assessed by liquid chromatography-tandem mass spectrometry.

BACKGROUND: Men with Klinefelter syndrome (KS) show hypergonadotropic hypogonadism, but the pathogenesis of hypotestosteronemia remains unclear. Testicular steroidogenesis in KS men was evaluated over three decades ago after human chorionic gonadotropin (hCG) stimulation, but inconclusive results were obtained. Intriguingly, some recent studies show increased intratesticular testosterone concentrations in men with KS. OBJECTIVE: To analyze serum steroid profile, as a proxy of testicular steroidogenesis, after hCG stimulation in KS compared with control men. DESIGN: A prospective, longitudinal, case-control, clinical trial. METHODS: Thirteen KS patients (36±9 years) not receiving testosterone (TS) replacement therapy and 12 eugonadic controls (32±8 years) were enrolled. Serum steroids were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) at baseline and for five consecutive days after intramuscular injection of 5000IU hCG. RESULTS: Progesterone (P), 17-hydroxyprogesterone (17OHP), TS, and estradiol (E2) showed a significant increase (P<0.001) after hCG stimulation in both groups. On the contrary, androstenedione (AS) and dehydroepiandrosterone did not increase after hCG stimulation. The 17OHP/P ratio increased in both groups (P<0.001), the TS/AS ratio (17ß-hydroxysteroid dehydrogenase type 3 (17ßHSD3) activity) did not increase after hCG in any group, and the E2/TS ratio (aromatase activity) increased significantly in both groups (P=0.009 in KS and P<0.001 in controls). Luteinizing hormone decreased after hCG in both groups (P=0.014 in KS and P<0.001 in controls), whereas follicle-stimulating hormone decreased only in control men (P<0.001). CONCLUSION: This study demonstrates for the first time using LC-MS/MS that Leydig cells of KS men are able to respond to hCG stimulation and that the first steps of steroidogenesis are fully functional. However, the TS production in KS men is impaired, possibly related to reduced hydroxysteroid deydrogenase activity due to an unfavorable intratesticular metabolic state.

Effect of a single oral dose of oxymetholone on the metabolism of human erythrocytes.

Androgenic steroids have been shown to enhance erythrocyte 2,3-DPG production in vivo and in vitro, and to stimulate the pentose shunt oxidative reactions in vitro. Furthermore, a 3 beta- and a 17 beta-hydroxysteroid dehydrogenase have been identified in red cells. The present study was carried out to explore a cumulative effect of androgens on glycolysis and androgen reduction in human erythrocytes in vivo following a single 50 mg oral dose of 17 beta-hydroxy-2 (hydroxymethylene)-17 methyl-5 alpha-androstan-3-one (oxymetholone). The rate of erythrocyte glycolysis was measured by quantitative determination of: fructose-1,6-diphosphate (FDP); dehydroxyacetone phosphate (DAP); 2,3-diphosphoglycerate (2,3-DPG); adenosine triphosphate (ATP); and lactate. Serum and erythrocyte steroids were separated by thin layer chromatography. The reduction of 5 alpha-androstan-17 beta-ol-3-one by red cell hemolysate was measured in the presence of NADPH as an index of 3(17)beta-hydroxysteroid dehydrogenase activity. Our results show that oxymetholone administration is followed by the appearance of an unidentified steroid fraction in chromatograms of serum and erythrocytes, simultaneously with the enhancement of glycolysis and of hydroxysteroid dehydrogenase activity in erythrocytes. A direct effect of androgen on erythrocyte metabolism, which is independent of the hormone erythropoietic effect, is discussed.

Diabetes and alcohol: Double jeopardy with regard to oxidative toxicity and sexual dysfunction in adult male Wistar rats.

The aim of this study was to test whether diabetic rats exposed to alcohol demonstrate a higher degree of reproductive toxicity and suffer with elevated oxidative toxicity when compared with alcohol exposed control rats. Diabetes was induced by injecting single dose of streptozotocin and alcohol was administered through orogastric tube once daily for a period of 55 days. Daily sperm production, epididymal sperm count, motile, viable and HOS-tail coiled sperms, serum testosterone levels and testicular 3ß- and 17ß-hydroxysteroid dehydrogenase activity levels were significantly decreased in diabetic rats. Significant reduction in testicular and epididymal superoxide dismutase and catalase activity levels, and elevation in lipid peroxidation products were observed in diabetic rats. Similar reproductive and oxidative toxicity was observed in alcohol treated control rats. Further, alcohol exposed diabetic rats showed additional deterioration in reproductive endpoints and noteworthy elevation in oxidative toxicity suggesting that treatment with alcohol further deteriorates sexual dysfunction in STZ-induced diabetic rats.

Type II estrogen-binding sites and 17 beta-hydroxysteroid dehydrogenase activity in human peripheral blood mononuclear cells.

Type II estrogen-binding sites (type II EBS) have been demonstrated in human peripheral blood mononuclear cells (PBMC) using a whole cell assay with [6,7-3H]estradiol [( 3H]E2) as tracer. During whole cell incubations for 60 min at 37 C for type II EBS quantification, we found that PBMC contain 17 beta-hydroxysteroid dehydrogenase (17 beta HSD) activity, which led to errors in estimating type II EBS concentrations by diminishing, by about 70%, the amount of available labeled E2. On the other hand, after 150 min at 4 C only 16% of the tracer was converted to estrone. Thus, we measured the maximal steady state binding in PBMC by incubating the cells with [3H]E2 at 4 C for 150 min. Equilibrium binding analysis of PBMC yielded sigmoid saturation curves with a saturation point at a ligand concentration of about 40 nmol/L. Scatchard analysis of binding data yielded a concave plot, which together with a Hill coefficient of 2.13, suggests that the type II EBS may have multiple binding sites which display positive cooperativity. The apparent equilibrium dissociation constant (Kd), determined from the [3H]E2 concentration required for half-saturation, was about 22 nmol/L. The type II EBS were estrogen specific, as demonstrated by competition experiments. Only those steroids with estrogenic activity inhibited binding of [3H]E2; nonestrogenic steroids did not. The type II EBS were found to be 3S macromolecules based on analysis of postlabeled fractions prepared by sucrose density gradient centrifugation. The number of type II EBS in PBMC from normal women was highest during the late follicular-early luteal phase of the menstrual cycle. We conclude that human PBMC specifically take up, retain, and metabolize E2.

The longitudinal study of adrenal maturation during gonadal suppression: evidence that adrenarche is a gradual process.

The physical changes that herald the onset of puberty result from the combination of adrenarche and gonadarche. To examine adrenal maturation and associated changes in growth without the confounding effects of changes in the gonadal steroid milieu, we performed a longitudinal study in 14 young girls with idiopathic central precocious puberty during long-term pituitary-gonadal suppression. Beginning at the mean age of 2.9 yr, dehydroepiandrosterone sulfate levels, linear growth, skeletal maturation, body mass index, and secondary sexual development were evaluated at 3- to 6-month intervals for up to 12.3 yr. In 12 of the girls, levels of dehydroepiandrosterone, androstenedione, 17-hydroxypregnenolone, and 17alpha-hydroxyprogesterone were determined before and after acute ACTH stimulation every 6 months to investigate the maturation of adrenal steroidogenic enzyme activity. Serum dehydroepiandrosterone sulfate levels rose progressively throughout the study. An exponential model fit the longitudinal datasets well and indicated that dehydroepiandrosterone sulfate levels increased approximately 22%/yr from the youngest age onward. Increasing activity of 17-20 lyase (CYP17) and decreasing activity of 3beta-hydroxysteroid dehydrogenase were also evident in preadrenarchal subjects. When controlled for chronological age, no significant associations were noted between weight, body mass index, or body surface area and dehydroepiandrosterone sulfate levels. However, similar analyses revealed modest correlations of both height and growth velocity with dehydroepiandrosterone sulfate levels. Our results suggest that adrenarche is not the result of sudden rapid changes in adrenal enzyme activities or adrenal androgen concentrations; rather, adrenarche may be a gradual maturational process that begins in early childhood.

Salt loading affects cortisol metabolism in normotensive subjects: relationships with salt sensitivity.

We studied cortisol metabolism together with insulin sensitivity [homeostatic model assessment (HOMA)] and renal hemodynamics in 19 salt-resistant (sr) and nine salt-sensitive (ss) normotensive subjects after a low- and high-salt diet. Results are described as high- vs. low-salt diet. Sum of urinary cortisol metabolite excretion (sum(metabolites)) increased in sr subjects (3.8 +/- 1.6 vs. 3.1 +/- 1.1 microg/min per square meter, P < 0.05) and decreased in ss subjects (2.3 +/- 1.0 vs. 2.9 +/- 1.1 microg/min per square meter, P < 0.05). Plasma 0830 h cortisol decreased in sr subjects but did not change significantly in ss subjects. In all subjects, the absolute blood pressure change correlated negatively with the percentage change in sum(metabolites) (P < 0.05) and positively with the percentage change in renal vascular resistance (P < 0.05). Sum(metabolites) during high-salt diet correlated negatively with the percentage changes in plasma 0830 h cortisol (P < 0.05) and renal vascular resistance (P = 0.05). HOMA did not change in either group, but the percentage change in HOMA correlated positively with the percentage change in plasma cortisol (P = 0.001) and negatively with the percentage change in sum(metabolites) (P < 0.01). Parameters of 11 beta-hydroxysteroid dehydrogenase activity were not different between groups and did not change. In conclusion, these data suggest that cortisol elimination is affected differently after salt loading in sr and ss subjects. Changes in circulating cortisol might contribute to individual sodium-induced alterations in insulin sensitivity.

Luteal 3beta-hydroxysteroid dehydrogenase and 20alpha-hydroxysteroid dehydrogenase activities in the rat corpus luteum of pseudopregnancy: effect of the deciduoma reaction.

BACKGROUND: In the rat, the maintenance of gestation is dependent on progesterone production from the corpora lutea (CL), which are under the control of pituitary, decidual and placental hormones. The luteal metabolism of progesterone during gestation has been amply studied. However, the regulation of progesterone synthesis and degradation during pseudopregnancy (PSP), in which the CL are mainly under the control of pituitary prolactin (PRL), is not well known. The objectives of this investigation were: i) to study the luteal metabolism of progesterone during PSP by measuring the activities of the enzymes 3beta-hydroxysteroid dehydrogenase (3betaHSD), involved in progesterone biosynthesis, and that of 20alpha-hydroxysteroid dehydrogenase (20alphaHSD), involved in progesterone catabolism; and ii) to determine the role of decidualization on progesterone metabolism in PSP. METHODS: PSP was induced mechanically at 10:00 h on the estrus of 4-day cycling Wistar rats, and the stimulus for decidualization was provided by scratching the uterus on day 4 of PSP. 3betaHSD and 20alphaHSD activities were measured in the CL isolated from ovaries of PSP rats using a spectrophotometric method. Serum concentrations of progesterone, PRL, androstenedione, and estradiol were measured by radioimmunoassay (RIA). RESULTS: The PSP stage induced mechanically in cycling rats lasted 11.3 +/- 0.09 days (n = 14). Serum progesterone concentration was high until day 10 of PSP, and declined thereafter. Serum PRL concentration was high on the first days of PSP but decreased significantly from days 6 to 9, having minimal values on days 10 and 11. Luteal 3betaHSD activities were elevated until day 6 of PSP, after which they progressively declined, reaching minimal values at the end of PSP. Luteal 20alphaHSD activities were very low until day 9, but abruptly increased at the end of PSP. When the deciduoma was induced by scratching the uterus of pseudopregnant animals on day 4 (PSP+D), PSP was extended to 18 +/- 2.2 days (n = 8). In PSP + D rats, serum progesterone and PRL levels, and luteal 3betaHSD activities were higher than in pseudopregnant rats on day 11. Decidualization also prevented the increase in luteal 20alphaHSD activities observed on day 11 of PSP. Administration of the dopaminergic agonist CB154 in PSP + D rats on day 10 of PSP induced a decline in both serum PRL and progesterone on day 11 of PSP, values that were not different from that of pseudopregnant controls. CONCLUSIONS: We have established that during the final period of PSP a decline in progesterone biosynthesis occurs before the increase in progesterone catabolism. We have also shown that decidualization in pseudopregnant rats extends the life of the CL by prolonging the production of pituitary PRL, and by maintaining high 3betaHSD and low 20alphaHSD activities within the CL leading to sustained production of progesterone.

Low dose ketoconazole attenuates serum androgen levels in patients with polycystic ovary syndrome and inhibits ovarian steroidogenesis in vitro.

OBJECTIVE: To investigate the effects of a low-dose ketoconazole on ovarian steroidogenesis and on serum androgen levels in polycystic ovary syndrome (PCOS). DESIGN: In vitro, human granulosa-luteal cells were incubated with ketoconazole and radiolabeled steroid substrates, to follow their metabolic fate by thin-layer chromatography analysis. In vivo, normally cycling women (n = 7) in their luteal phase were administered one tablet of 200 mg ketoconazole at 8 A.M. Serum steroid levels, sampled basally and at 12 P.M., 4 P.M., and 8 A.M. the next morning, were compared with untreated control group (n = 7) values. Polycystic ovary syndrome women (n = 11) were similarly administered ketoconazole 6 to 10 days after occurrence of spontaneous menses. Adrenal origin of hyperandrogenemia was excluded by stimulation with ACTH and a normal basal DHEAS. The steroid diurnal variation was determined in the same patients a day before treatment. RESULTS: In vitro, ketoconazole selectively inhibited the key steroidogenic cytochromes, namely P450scc, P45017 alpha, and P450arom (IC50 = 0.5 to 1.0 microgram/mL). In vivo, in the luteal phase, ketoconazole transiently decreased serum values (mean +/- SE) of E2 (19.2% +/- 2.1%) and P (38.3% +/- 8.5%) within 4 to 8 hours. The same low-dose ketoconazole, administered to PCOS women, decreased serum values of androstenedione (17.6% +/- 4.7%), T (24.6% +/- 7.6%), and free T (30.7% +/- 7.7%). In contrast, 17 alpha-hydroxyprogesterone increased concomitantly (78.5% +/- 10.8%), suggesting a greater suppressibility of the P45017 alpha lyase activity. The E2 levels in PCOS patients were slightly elevated (29.1% +/- 5.6%), resulting in a 1.7- to 2.3-fold increase of the E2:T ratio. CONCLUSIONS: These findings suggest that a low-dose ketoconazole may facilitate a decreased intraovarian T:E2 ratio, which may prove favorable for follicular maturation in PCOS.

Effects of passive immunization against alpha-2u-globulin and supplementation with alpha-2u-globulin on serum gonadotrophins and testicular activity.

The purpose of the present study was to evaluate the effects of anti-alpha-2U-globulin on pituitary-gonadal functions in male rats. Adult male rats were given injections of anti-alpha-2u-globulin (1 mg/day) for 14 days, when they were killed 7 days after the last injection, serum levels of gonadotrophins and testosterone measured by radioimmunoassays, were less, testicular delta 3 beta- and 17 beta-hydroxy-steroid dehydrogenase (3 beta- and 17 beta-HSD) activities were suppressed, spermatogenesis was inhibited and serum level of alpha-2u-globulin was decreased in anti-alpha-2u-globulin treated rats. Administration of alpha-2u-globulin (1.5 mg/day) for 7 days to anti-alpha-2u-globulin treated rats reversed the 3beta-HSD and 17beta-HSD activities and serum levels of gonadotrophins, testosterone and alpha-2u-globulin, while spermatogenesis was restored to normal. The results indicate that changes in testicular steroidogenesis and spermatogenesis in rats after passive immunization against alpha-2u-globulin may be due to decrease in availability of endogenous alpha-2u-globulin.

Effect of continuous light on spermatogenesis and testicular steroidogenesis in rats: possible involvement of alpha 2u-globulin.

Male rats exposed to continous light for 70 days showed an increased weights of testis, accessory sex organs, serum levels of FSH, LH, testosterone, testicular 17beta-hydroxysteroid dehydrogenase (1beta-HSD) activity and alpha 2u-globulin, while 3beta-hydroxysteroid dehydrogenase activity showed no significant changes. Prolonged light exposure also stimulated sepermatogenesis in rats. These results suggest that alpha 2u-globulin possibly stimutates the male gonad by inducing pituitary gonadotropins in continuous light-exposed rats.