11β-HSD1 Activity Research Articles

11β-hydroxysteroid dehydrogenase type 1 is associated with antiretroviral therapy-induced increase in low-density lipoprotein cholesterol .

To investigate the association between 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) activity and antiretroviral therapy (ART)-induced increase in low-density lipoprotein cholesterol (LDL). We enrolled 62 patients and used liquid chromatography-tandem mass spectrometry to measure 11β-HSD1 activity, which was expressed as a ratio of the sum of urinary tetrahydrocortisol and allo-tetrahydrocortisol concentrations to urinary tetrahydrocortisone concentration. Patient data, including baseline laboratory values, were extracted from medical records for logistic regression analyses of factors associated with LDL increase during ART. The cutoff 11β-HSD1 activity ratio associated with the LDL increase during ART was determined using receiver operator characteristic (ROC) curve analysis. The LDL level increased significantly from 88.8mg/dL before ART to 106.7mg/dL during ART (p=0.04). Additionally, patients with increased LDL tended to have a higher 11β-HSD1 activity ratio (1.59 vs. 1.21, p=0.06) and longer duration of ART (13.9 vs. 10.2 months, p=0.07) than patients with unchanged or decreased LDL. The cutoff 11β-HSD1 activity ratio was 1.226. Results of the univariate logistic regression analysis suggested that 11β-HSD1 activity ratio ≥1.226 was associated with LDL increase during ART (p=0.011), with an odds ratio of 8.000. This study revealed the possible association between 11β-HSD1 activity and ART-induced LDL increase. The findings of this study suggest that 11β-HSD1 could be a useful drug target for the treatment of ART-induced hyperlipidemia.

2031-P: Adipose-Specific Lack of Hexose-6-Phosphate Dehydrogenase Causes Inactivation of Adipose Glucocorticoids and Improves Metabolic Phenotype in Male Mice

Excessive glucocorticoids (GCs) production in adipose tissue promotes the development of visceral obesity and the metabolic syndrome. 11β-Hydroxysteroid dehydrogenase type 1 (11 β-HSD1) is critical for controlling intracellular GC production in adipose tissue, and this process is also regulated by hexose-6-phosphate dehydrogenase (H6PDH) that provides cofactor NADPH for 11β-HSD1 amplifying local GC production. To better understand the integrated molecular and physiological role of adipose H6PDH, we created a tissue specific knockout of H6PDH gene mouse model in adipocytes (H6PDHAcKO mice) and characterized the metabolic related phenotypes. H6PDHAcKO mice had decreased levels of intra-adipose corticosterone with reduction of 11β-HSD1 activity in response to H6PDH deletion in adipose tissue. Moreover, H6PDHAcKO mice also exhibited reduced fasting blood glucose levels and increased glucose tolerance with increased insulin sensitivity. In addition, plasma FFA levels were decreased with a concomitant decrease in the expression of lipase ATGL and HSL in the abdominal adipose tissue. These results indicate that inactivation of H6PDH expression in adipose tissue is sufficient to cause intra-adipose GC inactivation leading to the favorable pattern of metabolic phenotype. These data suggest that H6PDHAcKO mice may provide a good model for study the potential contributions of fat-specific H6PDH inhibition in the metabolic phenotype and other pathological disorders in vivo. Disclosure J. Wang: None. X. Wang: None. K. Lutfy: None. T. Friedman: None. M. Jiang: None. Y. Liu: None. Funding National Institute of Diabetes and Digestive and Kidney Diseases (SC1DK104821)

Steroid metabolomic signature of liver disease in nonsyndromic childhood obesity.

ObjectiveAnalysis of steroids by gas chromatography-mass spectrometry (GC-MS) defines a subject’s steroidal fingerprint. Here, we compare the steroidal fingerprints of obese children with or without liver disease to identify the ‘steroid metabolomic signature’ of childhood nonalcoholic fatty liver disease.MethodsUrinary samples of 85 children aged 8.5–18.0 years with BMI >97% were quantified for 31 steroid metabolites by GC-MS. The fingerprints of 21 children with liver disease (L1) as assessed by sonographic steatosis (L1L), elevated alanine aminotransferases (L1A) or both (L1AL), were compared to 64 children without markers of liver disease (L0). The steroidal signature of the liver disease was generated as the difference in profiles of L1 against L0 groups.ResultsL1 comparing to L0 presented higher fasting triglycerides (P = 0.004), insulin (P = 0.002), INS/GLU (P = 0.003), HOMA-IR (P = 0.002), GGTP (P = 0.006), AST/SGOT (P = 0.002), postprandial glucose (P = 0.001) and insulin (P = 0.011). L1AL showed highest level of T-cholesterol and triglycerides (P = 0.029; P = 0.044). Fasting insulin, postprandial glucose, INS/GLU and HOMA-IR were highest in L1L and L1AL (P = 0.001; P = 0.017; P = 0.001; P = 0.001). The liver disease steroidal signature was marked by lower DHEA and its metabolites, higher glucocorticoids (mostly tetrahydrocortisone) and lower mineralocorticoid metabolites than L0. L1 patients showed higher 5α-reductase and 21-hydroxylase activity (the highest in L1A and L1AL) and lower activity of 11βHSD1 than L0 (P = 0.041, P = 0.009, P = 0.019).ConclusionsThe ‘steroid metabolomic signature’ of liver disease in childhood obesity provides a new approach to the diagnosis and further understanding of its metabolic consequences. It reflects the derangements of steroid metabolism in NAFLD that includes enhanced glucocorticoids and deranged androgens and mineralocorticoids.

Serum and tissue pregnanes and pregnenes after dexamethasone treatment of cows in late gestation.

Dexamethasone (DEX) initiates parturition by inducing progesterone withdrawal and affecting placental steroidogenesis, but the effects of DEX in fetal and maternal tissue steroid synthetic capacity remains poorly investigated. Blood was collected from cows at 270 days of gestation before DEX or saline (SAL) treatment, and blood and tissues were collected at slaughter 38 h later. Steroid concentrations were determined by liquid chromatography tandem mass spectrometry to detect multiple steroids including 5α-reduced pregnane metabolites of progesterone. The activities of 3β-hydroxysteroid dehydrogenase (3βHSD) in cotyledonary and luteal microsomes and mitochondria and cotyledonary microsomal 5α-reductase were assessed. Quantitative PCR was used to further assess transcripts encoding enzymes and factors supporting steroidogenesis in cotyledonary and luteal tissues. Serum progesterone, pregnenolone, 5α-dihydroprogesterone (DHP) and allopregnanolone (3αDHP) concentrations (all <5 ng/mL before treatment) decreased in cows after DEX. However, the 20α-hydroxylated metabolite of DHP, 20αDHP, was higher before treatment (≈100 ng/mL) than at slaughter but not affected by DEX. Serum, cotyledonary and luteal progesterone was lower in DEX- than SAL-treated cows. Progesterone was >100-fold higher in luteal than cotyledonary tissues, and serum and luteal concentrations were highly correlated in DEX-treated cows. 3βHSD activity was >5-fold higher in luteal than cotyledonary tissue, microsomes had more 3βHSD than mitochondria in luteal tissue but equal in cotyledonary sub-cellular fractions. DEX did not affect either luteal or cotyledonary 3βHSD activity but luteal steroidogenic enzyme transcripts were lower in DEX-treated cows. DEX induced functional luteal regression and progesterone withdrawal before any changes in placental pregnene/pregnane synthesis and/or metabolism were detectable.

Increased central adiposity and decreased subcutaneous adipose tissue 11β-hydroxysteroid dehydrogenase type 1 are associated with deterioration in glucose tolerance-A longitudinal cohort study.

Increasing adiposity, ageing and tissue-specific regeneration of cortisol through the activity of 11β-hydroxysteroid dehydrogenase type 1 have been associated with deterioration in glucose tolerance. We undertook a longitudinal, prospective clinical study to determine if alterations in local glucocorticoid metabolism track with changes in glucose tolerance. Sixty-five overweight/obese individuals (mean age 50.3±7.3years) underwent oral glucose tolerance testing, body composition assessment, subcutaneous adipose tissue biopsy and urinary steroid metabolite analysis annually for up to 5years. Participants were categorized into those in whom glucose tolerance deteriorated ("deteriorators") or improved ("improvers"). Deteriorating glucose tolerance was associated with increasing total and trunk fat mass and increased subcutaneous adipose tissue expression of lipogenic genes. Subcutaneous adipose tissue 11β-HSD1 gene expression decreased in deteriorators, and at study completion, it was highest in the improvers. There was a significant negative correlation between change in area under the curve glucose and 11β-HSD1 expression. Global 11β-HSD1 activity did not change and was not different between deteriorators and improvers at baseline or follow-up. Longitudinal deterioration in metabolic phenotype is not associated with increased 11β-HSD1 activity, but decreased subcutaneous adipose tissue gene expression. These changes may represent a compensatory mechanism to decrease local glucocorticoid exposure in the face of an adverse metabolic phenotype.

OR10-1 Regulation of Glucocorticoid Metabolism: A Novel Function for Thyroid Hormones?

Glucocorticoid hormone metabolism and action are regulated by the 11-beta hydroxysteroid dehydrogenase (11βHSD) isozymes: the 11βHSD2, mostly expressed in the distal nephron, converts cortisol [F] into cortisone [E] in humans or corticosterone [B] into 11-dehydrocorticosterone [A] in rodents (11-dehydro derivatives being inactive compounds), and the 11βHSD1, ubiquitously expressed but abundant in the liver, catalyzes the opposite reaction. Under pathophysiological conditions of severe hypothyroidism, altered glucocorticoid metabolism has been observed, with decreased [F] to [E] conversion1,2. However, direct functional relationship between these two hormonal pathways has never been demonstrated to date. Using bioinformatics analyses, we identified five thyroid hormone response elements in the promoter region of the murine hsd11b2 gene. Therefore, we aimed at investigating whether thyroid hormones (T3) directly regulate expression and/or activity of the 11βHSD2 enzyme. We used three complementary models: human and mouse translational studies and molecular analyses in HEK293T cells and in fully differentiated KC3AC1 cortical collecting duct cells. Children and adults in hypo- or hyperthyroidism status were first compared either to age- and sex-matched euthyroid controls, or to themselves after reaching a euthyroid status. The urinary [E]/[F] ratio measured by LC-MS/MS method, was used as an index of renal 11βHSD2 activity. Interestingly, a 60% decrease in the [E]/[F] ratio was observed in hypothyroid patients, corroborating our hypothesis (p<0.05). Next, a mouse model of hyperthyroidism, generated by administrating T3 in drinking water, led to a significant 10% increase in renal 11βHSD2 mRNA and protein levels compared to wild type mice (n=10, p<0.05). Furthermore, we demonstrated in HEK293T cells that T3 transactivates the mouse hsd11b2 promoter cloned upstream the luciferase reporter gene via the Thyroid Receptor α1(TRα1). Finally, we showed that T3 exposure induces a 50% increase in 11βHSD2 mRNA levels in KC3AC1 cells, in a dose-dependent and time-dependent manner (as early as 6 h) (p<0.01). This induction was abrogated by Actinomycin D or DRB, two inhibitors of transcription, underlining a transcriptional regulatory mechanism. In addition, 11βHSD2 enzymatic activity, quantified by the [F] to [E] conversion ratio measured by LC-MS/MS in cell supernatants, increased significantly by 20% (p<0.05) after 24 h exposure to 100 nM T3. ChIP experiments further demonstrated a T3-dependent specific recruitment of the TRα1 onto the hsd11b2 promoter region. Altogether, our findings constitute the first demonstration that thyroid hormones directly regulate expression and activity of the renal 11βHSD2 enzyme, thereby controlling glucocorticoid metabolism and action.

SAT-357 Cotton Swab-Based Metabolic Signatures of Salivary Cortisol Measured by Liquid Chromatography-Mass Spectrometry

The comprehensive screening assay of salivary cortisol and its metabolic analogs is necessary to evaluate their pathophysiological functions in various adrenal diseases, such as Cushing’s Syndrome (CS). In contrast to the conventional method with Salivette®, the cotton swab may provides a point-of-care sampling for clinical screening as well as emergency management, and it also takes an advantage of sample preparation protocol. Sample preparation of salivary cortisol and its metabolic analogs was achieved by purification with solid-phase extraction after enzymatic hydrolysis with β-glucuronidase, and the sample was quantitatively measured by liquid chromatography-mass spectrometry (LC-MS) with selective MS scan modes and polarity swithcing. Due to uncertain sample volume in the cotton swab, the metabolic ratio of cortisol to cortisone, corresponding for 11β-HSD activity, may be an alternative marker in addition to individual salivary concentrations of cortisol and its analogs. The metabolic ratios of cortisol to cortisone in control group was higher than those of CS patients (1.93±4.09 vs. 0.66±1.08), while the increased ratios from 0.63±0.99 to 1.26±1.24 were obtained after surgery. The present swab-based LC-MS method may be appropriate for collecting saliva and the metabolic ratios of cortisol to its analogs could be the good screening tool for adrenal diseases.

The 11β-hydroxysteroid dehydrogenase isoforms: pivotal catalytic activities yield potent C11-oxy C19 steroids with 11βHSD2 favouring 11-ketotestosterone, 11-ketoandrostenedione and 11-ketoprogesterone biosynthesis

The 11β-hydroxysteroid dehydrogenase (11βHSD) types 1 and 2 are primarily associated with glucocorticoid inactivation and reactivation. Several adrenal C11-oxy C19 and C11-oxy C21 steroids, which have been identified in prostate cancer, 21-hydroxylase deficiency and polycystic ovary syndrome, are substrates for these isozymes. This study describes the kinetic parameters of 11βHSD1 and 11βHSD2 towards the C11-keto and C11-hydroxy derivatives of the C19 and C21 steroids. The apparent Km and Vmax values indicate the more prominent 11βHSD2 activity towards 11β-hydroxy androstenedione, 11β-hydroxytestosterone and 11β-hydroxyprogesterone in contrast to the 11βHSD1 reduction of the C11-keto steroids, as was demonstrated in the LNCaP cell model in the production of 11-ketotestosterone and 11-ketodihydrotestosterone. Data highlighted the role of 11βHSD2 and cytochrome P450 17A1 in the contribution of C11-oxy C21 steroids to the C11-oxy C19 steroid pool in the C11-oxy backdoor pathway. In addition, 11βHSD2 activity, catalysing 11-ketotestosterone biosynthesis, was shown to be key in the production of prostate specific antigen and in the progression of prostate cancer to castration resistant prostate cancer. The study at hand thus provides evidence that 11βHSD isozymes play key roles in pathophysiological states, more so than was previously put forward.

Maternal Blood Pressure, Cord Glucocorticoids, and Child Neurodevelopment at 2 Years of Age: A Birth Cohort Study.

Pregnancy hypertensive disorders have impaired neurodevelopment in offspring. We aimed to explore the association of normal range maternal blood pressure (BP) with child neurodevelopment, as well as the possible role of placental 11-beta-hydroxysteroid dehydrogenase type 2 (11β-HSD2) therein. Among 1,008 mother-child pairs recruited in Wuhan, China, in 2013-2015, we measured maternal third-trimester BP (systolic BP (SBP) and diastolic BP (DBP)) and cord glucocorticoids (cortisol and cortisone), a marker reflecting placental 11β-HSD2 activity. We evaluated child neurodevelopment using the Bayley Scales of Infant Development (BSID) with obtaining the Mental and Psychomotor Development Index (MDI and PDI). Multiple regression and mediation analysis were performed to estimate the effect. Each 5 mm Hg increase in maternal third-trimester SBP was associated with 1.54 points decrease in MDI (95% confidence interval (CI) = -2.60, -0.48) and 1.23 points decrease in PDI (95% CI = -2.14, -0.31); similar association was observed between DBP and BSID (adjusted β = -1.32; 95% CI = -2.53, -0.10 for MDI and -1.37; 95% CI = -2.42, -0.33 for PDI). Also, we found significant associations between cord cortisol/cortisone ratio and PDI (adjusted β = 2.95; 95% CI = 0.91, 4.99), as well as between maternal BP and cord cortisol/cortisone ratio (adjusted β = -0.03; 95% CI = -0.06, -0.01 for both SBP and DBP). Mediation analysis revealed that cord cortisol/cortisone ratio explained 6.29% of the association between SBP and PDI, and 6.85% between DBP and PDI. Increased maternal normal range BP may affect child neurodevelopment. Furthermore, placental 11β-HSD2 activity might be involved in the process.

A Lower Maternal Cortisol-to-Cortisone Ratio Precedes Clinical Diagnosis of Preterm and Term Preeclampsia by Many Weeks.

ContextPrevious studies have shown reduced placental levels of 11-β-hydroxysteroid dehydrogenase type 2 (11βHSD2) in preeclampsia (PE). However, it is unknown if the maternal cortisol-to-cortisone ratio is predictive of placental complications of pregnancy.ObjectiveTo determine the relationship between the maternal serum cortisol-to-cortisone ratio at different stages of pregnancy and the risk of PE or fetal growth restriction (FGR).DesignWomen from the Pregnancy Outcome Prediction Study experiencing PE (n = 194) or FGR (n = 185), plus a random sample of healthy controls (n = 279), were studied. Steroids were measured at ∼12, ∼20, ∼28, and ∼36 weeks of gestational age (wkGA). Separate analyses were performed for outcomes with term or preterm delivery. Associations were modeled using logistic regression.ResultsAt 28 wkGA, the cortisol-to-cortisone ratio was negatively associated (OR per 1 SD increase, 95% CI)] with preterm PE (OR 0.33, 95% CI 0.19 to 0.57), term PE (OR 0.61, 95% CI 0.49 to 0.76), and preterm FGR (OR 0.50, 95% CI 0.29 to 0.85). At 36 wkGA, the cortisol-to-cortisone ratio was negatively associated with term PE (OR 0.42, 95% CI 0.32 to 0.55) but not term FGR (OR 1.07, 95% CI 0.87 to 1.31). Associations were not materially affected by adjustment for maternal characteristics.ConclusionsA lower maternal serum cortisol-to-cortisone ratio precedes clinical manifestation of PE and preterm FGR by many weeks, despite previous reports of reduced levels of placental 11βHSD2 in these conditions. Our observations implicate enhanced maternal 11βHSD2 activity or reduced 11βHSD type 1 activity in the pathophysiology of PE.

Cellular localization and seasonal variation in BMP15 expression in ovary of the catfish Clarias batrachus and its role in ovarian steroidogenesis

Despite important actions of BMP15 (a TGFβ superfamily member) in follicular development in vertebrates, studies are mostly limited to mammals. The folliculogenic processes in lower vertebrates, particularly, fishes are quite different from mammals. It was, therefore, decided to detect the presence of BMP15 in ovarian follicles and what are its role in folliculogenesis and steroidogenesis in catfish? BMP15 protein was detected in different cellular compartments of ovarian follicles of the catfish collected during different reproductive phases, using immunohistochemical and Western blotting methods, with concurrent measurement of ovarian steroids through ELISA. In vitro effects of rhBMP15 on ovarian steroids, expression and activities of steroidogenic enzymes and StAR were also analyzed using established immunoblotting and spectrophotometric methods. BMP15 was localized distinctly in the nest of oogonia, perinucleolar oocytes and in oocytes as well as follicular cells of the primary ovarian follicles, which started diminishing gradually with the progression of folliculogenesis in oocytes-II and finally it was greatly reduced in the oocytes-III (fully grown follicles). BMP15 expression in follicles showed negative correlation with ovarian steroid levels. Further BMP15 also inhibited steroids production by suppressing the expression and activities of ovarian 3β-HSD, 17β-HSD and aromatase, however, it did not influence the expression of StAR. The findings of study suggests that BMP15 help in maintaining the early-stage oocytes in catfish and inhibits follicular growth by reducing ovarian steroidogenesis through suppression of the expression and activities of steroidogenic enzymes.

Diet formulated with black seed flour activates regulatory androgenic enzymes and neuroendocrine hormones in leachate-exposed reproductive damage of rat testes

Mixed environmental toxicants particularly mixed-metals had been implicated as testicular endocrine disruptors. Functional foods were applied in traditional medicine since decades for the treatment of several reproductive defects with limited scientific basis. The finding examines the steroidogenic proteins and neuroendocrine hormones linked to spermatogenesis and testicular wellness when exposed to toxicant-liquid from Elewi-odo vehicle-battery recycling site (EOMABRL) in rat, following treatment with black seed flour. Rats were distributed into six assemblages (n = 8): normal rats; normal rats administered with 10% and 20% black seed flour (BSF), respectively; testicular exposed rat; exposed rats treated with 10% and 20% supplementation of BSF, respectively; for 14 days. EOMABRL intoxication inhibited Δ5 3β-HSD and Δ5 17β-HSD activities and declined FSH, LH and testosterone levels. Diet supplemented with BSF at 10% and 20% significantly (p < 0.05) increased testicular steroidogenic enzymes (Δ5-17β-HSD and Δ5-17β-HSD) as well neuroendocrine hormones (LH, FSH) and testosterone levels in EOMABRL-intoxicated rats. Also, BSF prevented the alterations associated with spermatogenesis by potentiating seminiferous tubules (STs) and spermatocytes. We therefore concluded that diet formulated with BSF triggers the production of testicular endocrine hormones and hypothalamus pituitary–testicular axis via resuscitating germ cells in compromised rats.

Prenatal maternal stress is associated with lower cortisol and cortisone levels in the first morning urine of 45-month-old children

Prenatal stress (PS) has been related to altered hypothalamic-pituitary-adrenal (HPA) axis activity later in life. So far, studies in children assessing HPA axis functioning have focused on salivary cortisol, reflecting daytime activity. The present work is part of a prospective study and aims to extend knowledge about the association between PS and HPA axis regulation in children.To do so, we investigated cortisol, cortisone, and the ratio cortisone/(cortisone + cortisol) in the first morning urine of 45-month-old children in relation to several measures of maternal stress during pregnancy. Urinary cortisol and cortisone were measured by online turbulent flow chromatography coupled with high performance liquid chromatography-tandem mass spectrometry. PS was defined as: perceived stress for aim 1 (Perceived Stress Scale; n = 280); presence of self-reported (n = 371) and expert-rated psychopathology for aim 2 (Mini International Neuropsychiatric Interview; n = 281); continuous measures of anxiety and depression for exploratory aim 3 (State-Trait Anxiety Inventory and Edinburgh Postnatal Depression Scale; n = 280).Aim 1: Perceived maternal PS showed negative associations with cortisol and cortisone levels. Aim 2: The presence of expert-rated maternal psychopathology was associated with reduced morning cortisone. Aim 3: Continuous measures of anxiety and depression showed negative associations with cortisol and cortisone levels. After correcting for multiple testing, perceived maternal PS (aim 1) and prenatal level of anxiety (aim 3) were significant predictors of children’s urinary cortisol and cortisone in the morning (and, in the case of cortisone, also prenatal level of depression).The ratio cortisone/(cortisone + cortisol) as a global marker for the balance between the enzymes metabolizing cortisol to cortisone and vice versa (11β-hydroxysteroid dehydrogenases type 1 and 2; 11β-HSD1 and 2) was not associated with any measure of maternal PS (aims 1–3).The present study provides insight into possible programming effects of PS on nocturnal HPA axis activity and a proxy of 11β-HSD in a large sample. The results suggest that the nocturnal rate of cortisol production is lower in children exposed to PS, but do not support the hypothesis of divergent 11β-HSD activity.

Clomiphene citrate ameliorated lead acetate-induced reproductive toxicity in male Wistar rats.

Objective:The current study investigated the effects of clomiphene citrate on the hypothalamic-pituitary-testicular axis, steroidogenesis, sperm parameters, and testicular antioxidant enzyme activity of male Wistar rats submitted to lead acetate (Pb)-induced reproductive toxicity. Methods:Twenty adult male Wistar rats were divided into four groups of equal size as follows: Control; Clomid (0.35 mg/kg); Pb (10 mg/kg); and Clomid + Pb. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone, testicular 17-β hydroxysteroid dehydrogenase (17-β HSD) activity, androgen receptors, catalase activity, superoxide dismutase (SOD), malondialdehyde (MDA), sperm motility, viability, counts and morphology were estimated after oral administration of Clomid and/or lead acetate for 35 consecutive days. Data were analyzed using ANOVA at p<0.05.Results:Lead acetate significantly decreased (p<0.05) serum LH and testosterone levels, testicular 17β-HSD activity, androgen receptor expression, sperm motility, viability, counts, catalase activity, and SOD when compared with controls. Abnormal sperm morphology and MDA were significantly increased (p<0.05) in the Pb group compared with controls. Clomid co-administrated with lead acetate significantly increased (p<0.05) serum LH, testosterone levels, testicular 17β-HSD, androgen receptor expression, sperm motility and viability when compared with the group given lead acetate.Conclusions:The present study suggests that clomiphene citrate may stimulate testicular testosterone synthesis, sperm motility and viability via luteinizing hormone in a context of lead acetate-induced reproductive toxicity.

Down-regulation of steroidogenesis-related genes and its accompanying fertility decline in streptozotocin-induced diabetic male rats: ameliorative effect of metformin.

Metformin has long been used for glycemic control in diabetic state. Recently, other benefits of metformin beyond blood glucose regulation have emerged. To investigate the effect of metformin on the expression of testicular steroidogenesis-related genes, spermatogenesis, and fertility of male diabetic rats. Eighteen adult male Sprague Dawley rats were divided into three groups, namely normal control (NC), diabetic control (DC), and metformin-treated (300mg/kg body weight/day) diabetic rats (D+Met). Diabetes was induced using a single intraperitoneal injection of streptozotocin (60mg/kg b.w.), followed by oral treatment with metformin for four weeks. Diabetes decreased serum and intratesticular testosterone levels and increased serum but not intratesticular levels of luteinizing hormone. Sperm count, motility, viability, and normal morphology were decreased, while sperm nuclear DNA fragmentation was increased in DC group, relative to NC group. Testicular mRNA levels of androgen receptor, luteinizing hormone receptor, cytochrome P450 enzyme (CYP11A1), steroidogenic acute regulatory (StAR) protein, 3β-hydroxysteroid dehydrogenase (HSD), and 17β-HSD, as well as the level of StAR protein and activities of CYP11A1, 3β-HSD, and 17β-HSD, were decreased in DC group. Similarly, decreased activities of epididymal antioxidant enzymes and increased lipid peroxidation were observed in DC group. Consequently, decreased litter size, fetal weight, mating and fertility indices, and increased pre- and post-implantation losses were recorded in DC group. Following intervention with metformin, we observed increases in serum and intratesticular testosterone levels, Leydig cell count, improved sperm parameters, and decreased sperm nuclear DNA fragmentation. Furthermore, mRNA levels and activities of steroidogenesis-related enzymes were increased, with improved fertility outcome. Diabetes mellitus is associated with dysregulation of steroidogenesis, abnormal spermatogenesis, and fertility decline. Controlling hyperglycemia is therefore crucial in preserving male reproductive function. Metformin not only regulates blood glucose level, but also preserves male fertility in diabetic state.

Maternal stress and placental function, a study using questionnaires and biomarkers at birth.

BackgroundPrenatal stress affects the health of the pregnant woman and the fetus. Cortisol blood levels are elevated in pregnancy, and fetal exposure to cortisol is regulated by the placenta enzyme 11β-HSD2. A decrease in enzyme activity allows more maternal cortisol to pass through the placental barrier. Combining the fetal and maternal cortisol to cortisone ratio into the adjusted fetal cortisol exposure (AFCE) represents the activity of the enzyme 11β-HSD2 in the placenta.AimTo investigate the effect of prenatal maternal stress on the ratio of cortisol and cortisone in maternal and fetal blood at birth in a normal population.MethodMaternal self-reported stress was assessed at one time-point, as late in the pregnancy as convenient for the participant, using the Depression Anxiety Stress Scales (DASS-42), Pregnancy Related Anxiety (PRA), and Major Life Events during pregnancy. The study included 273 participants from Copenhagen University Hospital. Maternal and umbilical cord blood was sampled directly after birth and cortisol and cortisone concentrations were quantified using UPLC chromatography. Data were analyzed in a five-step regression model with addition of possible confounders. The primary outcome was AFCE, and plasma concentrations of maternal and fetal cortisol and cortisone were secondary outcomes.ResultsSignificant associations were seen for the primary outcome AFCE and the plasma concentrations of maternal cortisol and fetal cortisone with exposure to Pregnancy Related Anxiety (PRA), though the associations were reduced when adjusting for birth related variables, especially delivery mode. The weight of the placenta affected the associations of exposures on AFCE, but not plasma concentrations of cortisol and cortisone in mother and fetus. Moreover, the study demonstrated the importance of delivery mode and birth strain on cortisol levels right after delivery.ConclusionOur main finding was associations between PRA and AFCE, which shows the effect of maternal stress on placental cortisol metabolism.

Differential Regulation of 11β-Hydroxysteroid Dehydrogenase Type 1 Activity in Patients with Differing Etiologies of Hypopituitarism

Pituitary patients with different etiologies of hypopituitarism exhibit differing phenotypes, despite similar replacement therapy strategies. We hypothesized that differential regulation of the isoenzyme 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1), which mediates the net autocrine conversion of cortisone to cortisol in adipose tissues and liver, may play a role. We studied 11β-HSD1 activity (using urine cortisol/cortisone metabolites ratio) in 36 hypopituitary patients with treated craniopharyngiomas, treated remitted Cushing disease, and treated nonfunctioning pituitary adenomas + prolactinomas on and off growth hormone (GH) replacement. 11β-HSD1 activity was higher in subjects with craniopharyngioma both on and off GH, as evidenced by increased tetrahydrocortisol to tetrahydrocortisone metabolite ratios compared to other diagnostic groups, but there was no difference in body mass index, insulin levels, serum hormone measurements, or hydrocortisone dose between groups. Craniopharyngiomas are associated with enhanced 11β-HSD1 activity compared to other diagnostic hypopituitary groups, and this may contribute to the adverse phenotypic and metabolic features seen in this condition. BMI = body mass index; Em = cortisone metabolites; Fm = cortisol metabolites; GH = growth hormone; 11β-HSD1 = 11β-hydroxysteroid dehydrogenase type 1; IGF-1 = insulin-like growth factor 1; NFPA = nonfunctioning pituitary adenoma; THE = tetrahydrocortisone; THF = tetrahydrocortisol.

A novel highly potent and selective 11β-hydroxysteroid dehydrogenase type 1 inhibitor, INU-101

11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) is a cortisol regenerating enzyme that amplifies tissue glucocorticoid levels, especially in the liver and adipose tissue. Knockout mice or a selective inhibitor of 11β-HSD1 improves metabolic syndrome parameters in preclinical models and human clinical trials. Here, we evaluated the therapeutic potential of INU-101, a potent and selective oral inhibitor of 11β-HSD1. The in vitro activity of 11β-HSD1 was measured using the homogeneous time-resolved fluorescence (HTRF) assay. Differentiated adipocytes were used to evaluate the cellular 11β-HSD1 activity. To determine the inhibitory effects on 11β-HSD1 in tissues, we performed ex vivo studies using liver and adipose tissue isolated from C57BL/6 J mice and Cynomolgus monkeys. KKAy mice, ob/ob mice and ZDF rats were administered INU-101 to evaluate whether this compound ameliorated metabolic abnormalities in obese and diabetic animals. INU-101 had highly potent inhibitory activity in mouse, monkey and human 11β-HSD1, derived from liver microsomes. The oral administration of INU-101 significantly inhibited 11β-HSD1 activity in the liver and adipose tissue of mice and monkeys. In KKAy mice, ob/ob mice and ZDF rats, the oral administration of INU-101 enhanced insulin sensitivity and lowered the fasting blood glucose level. Furthermore, INU-101 treatment decreased the body weight and ameliorated an improved lipid profile in the diabetic mouse model. These results suggest that the 11β-HSD1 inhibitor, INU-101 may serve as a novel drug candidate for the treatment of type 2 diabetes and metabolic syndrome.

Akebia saponin D reverses corticosterone hypersecretion in an Alzheimer’s disease rat model

BackgroundGlucocorticoid hormones are implicated in the pathogenesis of Alzheimer’s disease (AD) and other diseases including diabetes, hyperlipidemia, and osteoporosis. Akebia saponin D (ASD) possesses numerous pharmacological activities, including as an anti-AD, anti-hyperlipidemia, anti-diabetes, and anti-osteoporosis agent. The anti-AD effect of ASD is possibly through its regulation of glucocorticoid levels. PurposeThe present study was undertaken to investigate the neuroprotective effects of ASD on Aβ25-35-induced cognitive deficits and to elucidate its underlying mechanism of action. MethodsThe AD rat model was established by an intracerebroventricular injection of Aβ25-35 into the lateral ventricles. Spatial learning and anxiety state were assessed by Morris water maze task and elevated plus-maze assay, respectively. The degree of hypertrophy of adrenal gland was analyzed using the viscera coefficient. Corticosterone and ACTH concentrations in the plasm were measured using biochemical assay kits. The activity of 11β-hydroxysteroid dehydrogenase type-1 (11β-HSD1) in liver and groin fat pad was assessed by measuring cortisol production. ResultsCompared with the control group, AD rats displayed significant spatial learning and reference memory impairments, serious anxiety disorders, obvious hypertrophy of adrenal gland, elevated corticosterone and ACTH levels in the plasma, and increased 11β-HSD1 activity in liver and groin fat pad. ASD could significantly ameliorate the memory deficits and anxiety symptoms, markedly reduce the viscera coefficient of adrenal gland, observably decrease corticosterone and ACTH concentrations, and showed no effect on the activity of 11β-HSD1. ConclusionsThese results indicate that ASD might exert a significant neuroprotective effect on cognitive impairment, driven in part by reducing systemic corticosterone level by down-regulation of the hypothalamic-pituitary-adrenal (HPA) axis.

Thiazolo[3,2-a]pyrimidin-5-one derivatives as a novel class of 11β-hydroxysteroid dehydrogenase inhibitors

11β-hydroxysteroid type 1 dehydrogenase (11β-HSD1) is an enzyme that increases tissue concentrations of cortisol. Selective inhibitors of this enzyme regulate the level of cortisol and thus play a key role in the treatment of Cushing's syndrome, metabolic syndrome and type 2 diabetes.In this study the inhibitory activity of 29 thiazolo[3,2-a]pyrimidin-5-one derivatives on 11β-HSD1 were investigated. Studies were carried out with pooled human liver microsomes. A lot of analyzed compounds show activity for inhibiting 11β-HSD1 (up to 59.15% at concentration 10 µmol/l). Molecular docking simulation show that the molecule of the most active compound: 7-(cyclohexylmethyl)-2-iodomethyl-2,3-dihydrothiazolo[3,2-a]pyrimidin-5-one forms hydrogen bonds with Ala172, Leu171, Leu215 or Tyr177. In addition, the cycloalkane moiety can create the hydrophobic contacts with NADP+. For this compound also the most favourable Docking Score value was obtained.The most active compound only in the slight degree inhibits 11β-HSD2 activity and is a selective inhibitor of 11β-hydroxysteroid dehydrogenase type 1. Consequently it can have a real effect on the regulation of the cortisol level in the body.