Glucocorticoids (GCs) are hormones that are released in response to stressors and exhibit many activities, including immunomodulatory and anti-inflammatory activities. They are primarily synthesized in the adrenal gland but are also produced in peripheral tissues via regeneration of adrenal 11-oxo metabolites or by de novo synthesis from cholesterol. The present study investigated the influence of the microbiota on de novo steroidogenesis and regeneration of corticosterone in the intestine of germ-free (GF) and specific pathogen-free mice challenged with a physical stressor (anti-CD3 antibody i.p. injection). In the small intestine, acute immune stress resulted in increased mRNA levels of the proinflammatory cytokines IL1β, IL6 and Tnfα and genes involved in de novo steroidogenesis (Stard3 and Cyp11a1), as well as in regeneration of active GCs from their 11-oxo metabolites (Hsd11b1). GF mice showed a generally reduced transcriptional response to immune stress, which was accompanied by decreased intestinal corticosterone production and reduced expression of the GC-sensitive marker Fkbp5. In contrast, the interaction between stress and the microbiota was not detected at the level of plasma corticosterone or the transcriptional response of adrenal steroidogenic enzymes. The results indicate a differential immune stress-induced intestinal response to proinflammatory stimuli and local corticosterone production driven by the gut microbiota.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenases genetics   metabolism   MeSH
• Corticosterone metabolism   MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Mice MeSH
• Steroids metabolism   MeSH
• Gastrointestinal Microbiome physiology   MeSH
• Tandem Mass Spectrometry MeSH
• Intestine, Small metabolism   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

The commensal microbiota affects brain functioning, emotional behavior and ACTH and corticosterone responses to acute stress. However, little is known about the role of the microbiota in shaping the chronic stress response in the peripheral components of the hypothalamus-pituitary-adrenocortical (HPA) axis and in the colon. Here, we studied the effects of the chronic stress-microbiota interaction on HPA axis activity and on the expression of colonic corticotropin-releasing hormone (CRH) system, cytokines and 11β-hydroxysteroid dehydrogenase type 1 (11HSD1), an enzyme that determines locally produced glucocorticoids. Using specific pathogen-free (SPF) and germ-free (GF) BALB/c mice, we showed that the microbiota modulates emotional behavior in social conflicts and the response of the HPA axis, colon and mesenteric lymph nodes (MLN) to chronic psychosocial stress. In the pituitary gland, microbiota attenuated the expression of Fkbp5, a gene regulating glucocorticoid receptor sensitivity, while in the adrenal gland, it attenuated the expression of genes encoding steroidogenesis (MC2R, StaR, Cyp11a1) and catecholamine synthesis (TH, PNMT). The pituitary expression of CRH receptor type 1 (CRHR1) and of proopiomelanocortin was not influenced by microbiota. In the colon, the microbiota attenuated the expression of 11HSD1, CRH, urocortin UCN2 and its receptor, CRHR2, but potentiated the expression of cytokines TNFα, IFNγ, IL-4, IL-5, IL-6, IL-10, IL-13 and IL-17, with the exception of IL-1β. Compared to GF mice, chronic stress upregulated in SPF animals the expression of pituitary Fkbp5 and colonic CRH and UCN2 and downregulated the expression of colonic cytokines. Differences in the stress responses of both GF and SPF animals were also observed when immunophenotype of MLN cells and their secretion of cytokines were analyzed. The data suggest that the presence of microbiota/intestinal commensals plays an important role in shaping the response of peripheral tissues to stress and indicates possible pathways by which the environment can interact with glucocorticoid signaling.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 1 metabolism   MeSH
• Adrenocorticotropic Hormone metabolism   MeSH
• Behavior, Animal physiology   MeSH
• Cytokines metabolism   MeSH
• Gene Expression physiology   MeSH
• Glucocorticoids genetics   physiology   MeSH
• Corticotropin-Releasing Hormone metabolism   MeSH
• Pituitary Gland MeSH
• Corticosterone metabolism   MeSH
• Microbiota physiology   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Adrenal Glands MeSH
• Stress, Psychological genetics   metabolism   MeSH
• Psychology MeSH
• Receptors, Glucocorticoid metabolism   MeSH
• Gene Expression Regulation physiology   MeSH
• Social Behavior MeSH
• Pituitary-Adrenal System microbiology   MeSH
• Hypothalamo-Hypophyseal System microbiology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The bioavailability of glucocorticoids is modulated by enzyme 11β-hydroxysteroid dehydrogenase type 1 (11HSD1), which catalyzes the conversion of inactive 11-oxo-glucocorticoids to active 11-hydroxy-glucocorticoids cortisol and corticosterone and is regulated by pro-inflammatory cytokines. Our aim was to assess the effect of colitis on the expression of 11HSD1 in specific microanatomical compartments of the mucosal immune system. Using qRT-PCR we quantified the expression of 11HSD1 and cytokines in the colon, mesenteric lymph nodes (MLN) and spleen of mice with colitis. Microsamples of the MLN cortex, paracortex and medulla, colonic crypt epithelium (CCE), lamina propria and isolated intestinal lymphoid follicles (ILF) were harvested by laser microdissection, whereas splenic and MLN lymphocytes by flow cytometry. Colitis increased 11HSD1 in the CCE, ILF, and MLN cortex but not in the lamina propria and the MLN paracortex and medulla. Expression of IL-4, IL-21 and TNFα was increased in both the cortex of MLN and ILF, whereas IL-1β and IL-10 were only increased in the follicles. No positive effect was observed in the case of IFNγ and TGFβ. 11HSD1 was positively correlated with TNFα and less strongly with IL-21, IL-1β, and IL-4. Colitis also upregulated the 11HSD1 expression of T cells in the spleen and MLN. The study demonstrates the stimulatory effect of inflammation on local glucocorticoid metabolism only in particular compartments of the mucosal immune system. The correlation between cytokines and 11HSD1 in the ILF and MLN cortex indicates that pro-inflammatory cytokines may amplify glucocorticoid signals in inductive compartments of the mucosal immune system.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 1 genetics   metabolism   MeSH
• Cytokines metabolism   MeSH
• Colitis enzymology   genetics   immunology   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Gene Expression Regulation, Enzymologic MeSH
• Intestinal Mucosa immunology   MeSH
• Inflammation enzymology   genetics   immunology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Numerous chemicals in the environment have the ability to interact with the endocrine system. These compounds are called endocrine disruptors (EDs). Exposure to EDs represents one of the hypotheses for decreasing fertility, the increased risk of numerous cancers and obesity, metabolic syndrome and type 2 diabetes. There are various mechanisms of ED action, one of which is their interference in the action of 11β-hydroxysteroid dehydrogenase (11βHSD) that maintains a balance between active and inactive glucocorticoids on the intracellular level. This enzyme has two isoforms and is expressed in various tissues. Inhibition of 11βHSD in various tissues can have different consequences. In the case of EDs, the results of exposure are mainly adverse; on the other hand pharmaceutically developed inhibitors of 11βHSD type 1 are evaluated as an option for treating metabolic syndrome, as well as related diseases and depressive disorders. This review focuses on the effects of 11βHSD inhibitors in the testis, colon, adipose tissue, kidney, brain and placenta.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 1 antagonists & inhibitors   metabolism   MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 2 antagonists & inhibitors   metabolism   MeSH
• Diabetes Mellitus chemically induced   enzymology   pathology   MeSH
• Endocrine Disruptors pharmacology   MeSH
• Glucocorticoids metabolism   MeSH
• Enzyme Inhibitors pharmacology   MeSH
• Colon drug effects   enzymology   MeSH
• Humans MeSH
• Metabolic Syndrome chemically induced   enzymology   pathology   MeSH
• Brain drug effects   enzymology   MeSH
• Neoplasms chemically induced   enzymology   pathology   MeSH
• Obesity chemically induced   enzymology   pathology   MeSH
• Organ Specificity MeSH
• Placenta drug effects   enzymology   MeSH
• Pregnancy MeSH
• Testis drug effects   enzymology   MeSH
• Adipose Tissue drug effects   enzymology   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Endokrinní disruptor (ED) je definován jako exogenní látka, která zasahuje do syntézy, sekrece, transportu, vazby, akce nebo eliminace přirozených hormonů zodpovědných za udržování homeostázy, reprodukci, vývoj a/nebo chování. Cílem projektu je přispět k objasnění vlivu některých vybraných faktorů vnějšího prostředí na lidskou spermatogenezi, se zaměřením na steroidy a jejich úlohu v mužské reprodukci. Chceme vyhodnotit vliv vybraných ED – 6 druhů polychlorovaných bifenylů a bisfenolu A ze séra na kvalitu spermiogramu, hodnoty vybraných steroidů v seminální tekutině a v séru a na hodnoty homocysteinu a stopových prvků selenu a zinku v séru. K dispozici jsou 4 skupiny po 40 pacientech: mírně, středně a těžce neplodní muži a zdravý kontrolní soubor. Předpokládáme, že subfertilita u mírně a středně neplodných mužů by mohla být z velké části zapříčiněna expozicí endokrinním disruptorům. Projekt by mohl vnést více poznání do příčin poruch mužské reprodukce a přispět tak k její léčbě.; Endocrine disruptor (ED) is defined as an exogenous substance interfering with the synthesis, secretion, transport, binding or action of natural hormones responsible for reproduction, development and/or behavior. The project aspires to increase the knowledge about effects of selected ED on human spermatogenesis, focusing on steroids and their role in male reproduction. The aim is to evaluate the impact of selected ED - classes of polychlorinated biphenyls and bisphenol A from serum to determine spermiogram quality, levels of chosen steroids in the serum and seminal fluid and trace levels of selenium and zinc. 4 Groups of males, each consisting of 40 patients will be investigated: lightly, moderately and severely infertile males and a control group. We suppose that subfertility in lightly and moderately affected individuals could be, to a large extent, caused by an exposition to ED. Project will lead to the better understanding of the causes of male fertility disorders and contribute to the treatment.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenases MeSH
• Bisphenol A-Glycidyl Methacrylate MeSH
• Endocrine Disruptors MeSH
• Infertility, Male MeSH
• Polychlorinated Biphenyls MeSH
• Selenium MeSH
• Spermatogenesis MeSH
• Steroids MeSH
• Zinc MeSH

• Conspectus
• Fyziologie člověka a srovnávací fyziologie
• NML Fields
• endokrinologie
• reprodukční lékařství
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

Enzym 11beta HSD 1 je zapojen do procesů energetické bilance a příjmu potravy. U obézních dospívajících dětí ve věku 13-18 let budeme sledovat, jak se mění aktivita enzymu před léčbou a po 1 měsíci léčby. Zaměříme se na sérové hladiny především méně běžných steroidů, které jsou ukazateli enzymatické aktivity 11beta HSD 1. Měření budeme provádět rovněž u normostenických kontrol. K analýzám použijeme jinak nevyužitý materiál nasbíraný v jiném výzkumném projektu Endokrinologického ústavu zaměřeném na metabolický syndrom a dětskou obezitu. V projektu tedy nebude zapotřebí odebírat další biologické vzorky, čímž dojde k významnému snížení časových i finančních požadavků studie. Na základě naměřených dat se pokusíme objasnit roli enzymu 11beta HSD 1 v lidskéobezitě s možností následného využití inhibitorů tohoto enzymu k terapeutickým účelům.; The enzyme 11beta HSD 1 is involved in the processes of energy balance and food intake. We will investigate how the activity of the enzyme is changing before and after 1 month of treatment in obese adolescent patients. We will focus on the serum levels of especially less common steroids, which are indicators of enzymatic activity of 11beta HSD 1. Normostenic controls will be also investigated. For the analysis we will use otherwise unused biological material collected during another project of The Institute of Endocrinology focused on metabolic syndrome and childhood obesity. It will therefore not be necessary to collect other samples. Based on the measured data we will try to clarify the role of the enzyme 11beta HSD 1 in human obesity with the subsequent use of inhibitors of this enzyme for therapeutic purposes.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 1 antagonists & inhibitors   therapeutic use   MeSH
• Chromatography methods   utilization   MeSH
• Dehydroepiandrosterone analogs & derivatives   analysis   MeSH
• Energy Metabolism MeSH
• Adrenal Cortex Hormones MeSH
• Hydrocortisone analysis   MeSH
• Cortisone analysis   MeSH
• Metabolic Syndrome prevention & control   MeSH
• Pediatric Obesity MeSH

• Conspectus
• Pediatrie
• NML Fields
• pediatrie
• nutriční terapie, dietoterapie a výživa
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

The local concentration of glucocorticoids is intensively regulated by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD 1). Human 11beta-HSD 1 also reversibly catalyzes the inter-conversion of 7alpha-hydroxy- and 7beta-hydroxy-dehydroepiandrosterone (DHEA) into 7-oxo-DHEA. The cohort of 282 obese adolescents, 154 girls (median age 15.31 years, range 14.17-16.68 years) and 128 boys (median age 14.95 years, range 13.87-16.16 years), BMI (Body Mass Index) >90th percentile was examined. In samples collected before and after one month of reductive diet therapy, circulating levels of steroids were analyzed by liquid chromatography-tandem mass spectrometry and radioimmunoassay methods. The model of the treatment efficacy prediction was calculated. A significant reduction in circulating levels of cortisone, E2 and increased levels of 7beta-hydroxy-DHEA after the reductive treatment was observed. Levels of cortisol, DHEA, DHT sustained without any significant change. The predictive Orthogonal Projections to Latent Structures (OPLS) model explained 20.1 % of variability of BMI, z-score change by the basal levels of 7alpha-hydroxy-DHEA, DHEA, cortisol and E2 as the strongest predictors. Reduced levels of circulating cortisone and reduced ratios of oxygenated/reduced metabolites reflect increased reductase activity of 11beta-HSD 1 with reduced BMI, z-score. We hypothesize whether these changes can be attributed to the altered activity of 11beta-HSD 1 in the liver.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 1 blood   MeSH
• Biomarkers blood   MeSH
• Dehydroepiandrosterone blood   MeSH
• Liver metabolism   MeSH
• Cohort Studies MeSH
• Humans MeSH
• Adolescent MeSH
• Obesity blood   therapy   MeSH
• Diet, Reducing trends   MeSH
• Treatment Outcome MeSH
• Check Tag
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Dehydroepiandrosterone (DHEA) and its 7-oxo- and 7-hydroxy-metabolites occurring in the brain are considered neurosteroids. Metabolism of the latter is catalysed by 11β-hydroxysteroid dehydrogenase (11β-HSD) which also interconverts cortisol and cortisone. The concurrent metabolic reaction to DHEA 7-hydroxylation is the formation of 16α-hydroxy-DHEA. The LC-MS/MS method using triple stage quadrupole-mass spectrometer was developed for simultaneous quantification of free DHEA, 7α-hydroxy-DHEA, 7β-hydroxy-DHEA, 7-oxo-DHEA, 16α-hydroxy-DHEA, cortisol and cortisone in human plasma and cerebrospinal fluid (CSF). The method employs 500 μL of human plasma and 3000 μL of CSF extracted with diethyl ether and derivatized with 2-hydrazinopyridine. It has been validated in terms of sensitivity, precision and recovery. In plasma, the following values were obtained: limit of detection: 2-50p g/mL; limit of quantification: 5-140 pg/mL; within-day precision 0.58-14.58%; between-day precision: 1.24-13.89% and recovery: 85-113.2%). For CSF, the values of limit of detection: 2-28 pg/mL; limit of quantification: 6-94 pg/mL; within-day precision; 0.63-5.48%; between-day precision: 0.88-14.59% and recovery: 85.1-109.4% were acquired. Medians and concentration ranges of detected steroids in plasma and CSF are given in subjects with excluded normal pressure hydrocephalus (n=37; 65-80 years). The method enables simultaneous quantification of steroids important for the estimation of 11β-HSD activity in human plasma and CSF. It will be helpful in better understanding various degenerative diseases development and progression.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenases metabolism   MeSH
• Dehydroepiandrosterone analogs & derivatives   MeSH
• Immunologic Factors MeSH
• Humans MeSH
• Neurotransmitter Agents MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The aim of the present work was to study the influence of variable stress on the expression of 11β-hydroxysteroid dehydrogenase type 1 (11HSD1) and the neuropeptides corticotropin-releasing hormone (CRH), urocortins 2 and 3(UCN2, UCN3), arginine vasopressin (AVP), oxytocin (OXT) and adenylate cyclase-activating polypeptide (PACAP) in two inbred rat strains: stress hypo-responsive Lewis (LEW) and hyper-responsive Fisher 344 (F344) rats. We found site-specific and strain-dependent differences in the basal and stress-stimulated expression of 11HSD1, CRH, UCN2, UCN3 and PACAP. In LEW rats, stress upregulated 11HSD1 in the prefrontal cortex and lateral amygdala, whereas in F344 rats 11HSD1 was upregulated in the central amygdala and hippocampal CA2 and ventral but not dorsal CA1 region; no effect was observed in the paraventricular nucleus, pituitary gland and adrenal cortex of both strains. The expression of glucocorticoid receptors did not parallel the upregulation of 11HSD1. Stress also stimulated the expression of paraventricular OXT, CRH, UCN3 and PACAP in both strains but amygdalar CRH only in LEW and UCN2/UCN3 in F344 rats, respectively. The upregulation of PACAP and CRH was paralleled only by increased expression of PACAP receptor PAC1 but not CRH receptor type 1. These observations provide evidence that inbred F344 and LEW rats exhibit not only the well-known phenotypic differences in the activity of the HPA axis but also strain- and stress-dependent differences in the expression of genes encoding 11HSD1 and neuropeptides associated with the HPA axis activity. Moreover, the differences in 11HSD1 expression suggest different local concentration of corticosterone and access to GR in canonical and noncanonical structures of the HPA axis.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenase Type 1 genetics   metabolism   MeSH
• Amygdala metabolism   MeSH
• Arginine Vasopressin genetics   metabolism   MeSH
• Hippocampus metabolism   MeSH
• Corticotropin-Releasing Hormone genetics   metabolism   MeSH
• Pituitary Gland metabolism   MeSH
• Pituitary Adenylate Cyclase-Activating Polypeptide genetics   metabolism   MeSH
• Rats MeSH
• Adrenal Cortex metabolism   MeSH
• RNA, Messenger metabolism   MeSH
• Brain metabolism   MeSH
• Paraventricular Hypothalamic Nucleus metabolism   MeSH
• Oxytocin genetics   metabolism   MeSH
• Rats, Inbred F344 MeSH
• Rats, Inbred Lew MeSH
• Prefrontal Cortex metabolism   MeSH
• Stress, Psychological genetics   metabolism   MeSH
• Receptors, Glucocorticoid genetics   metabolism   MeSH
• Gene Expression Profiling MeSH
• Pituitary-Adrenal System metabolism   MeSH
• Hypothalamo-Hypophyseal System metabolism   MeSH
• Urocortins genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH

Primary adrenal insufficiency (PAI) is a rare condition in childhood which is either inherited (mostly) or acquired. It is characterized by glucocorticoid and maybe mineralocorticoid deficiency. The most common form in children is 21-hydroxylase deficiency, which belongs to the steroid biosynthetic defects causing PAI. Newer forms of complex defects of steroid biosynthesis are P450 oxidoreductase deficiency and (apparent) cortisone reductase deficiency. Other forms of PAI include metabolic disorders, autoimmune disorders and adrenal dysgenesis, e.g. the IMAGe syndrome, for which the underlying genetic defect has been recently identified. Newer work has also expanded the genetic causes underlying isolated, familial glucocorticoid deficiency (FGD). Mild mutations of CYP11A1 or StAR have been identified in patients with FGD. MCM4 mutations were found in a variant of FGD in an Irish travelling community manifesting with PAI, short stature, microcephaly and recurrent infections. Finally, mutations in genes involved in the detoxification of reactive oxygen species were identified in patients with unsolved FGD. Most mutations were found in the enzyme nicotinamide nucleotide transhydrogenase, which uses the mitochondrial proton pump gradient to produce NADPH. NADPH is essential in maintaining high levels of reduced forms of antioxidant enzymes for the reduction of hydrogen peroxide. Similarly, mutations in the gene for TXNRD2 involved in this system were found in FGD patients, suggesting that the adrenal cortex is particularly susceptible to oxidative stress.

• MeSH
• 11-beta-Hydroxysteroid Dehydrogenases deficiency   MeSH
• Adrenal Insufficiency diagnosis   etiology   metabolism   therapy   MeSH
• Child MeSH
• Glucocorticoids deficiency   MeSH
• Hirsutism complications   congenital   therapy   MeSH
• Infant MeSH
• Humans MeSH
• 46, XX Disorders of Sex Development complications   therapy   MeSH
• Child, Preschool MeSH
• Steroids biosynthesis   MeSH
• Steroid Metabolism, Inborn Errors complications   therapy   MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Child, Preschool MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH