RESUMEN
The metabolomics of urinary steroids was studied by gas chromatography-mass spectrometry in 25 patients with Cushing's syndrome without malignant potential and in 12 patients with malignant potential of adrenal neoplasms (Weiss score 1-3). Patients with adrenocortical adenoma (N=24) constituted the control group. In patients with Cushing's syndrome and malignant potential, increased urinary excretion of 16-oxo-androstendiol, tetrahydro-11-deoxycortisol, and 16-hydroxypregnendiol, which had 100% specificity and sensitivity >90% for the diagnosis of malignant potential. Additionally, non-classical 5-ene-pregnenes (16-OHpregnenolone, 21-OH-pregnenolone, 3ß,16,20-pregnentriol, and 3ß,17,20-pregnentriol) were identified. The revealed changes in the metabolomics of steroids can be early signs of malignant potential in patients with Cushing's syndrome. In patients with malignant potential, three signs of reduced activity of 11ß-hydroxysteroid dehydrogenase type 2 were detected and in patients without malignant potential, one sign was found. In patients with and without malignant potential, three signs increased activity of 5ß-reductase were found.
Asunto(s)
Neoplasias de las Glándulas Suprarrenales/diagnóstico , Adenoma Corticosuprarrenal/diagnóstico , Biomarcadores de Tumor/orina , Síndrome de Cushing/diagnóstico , Metabolómica/métodos , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 2/orina , Neoplasias de las Glándulas Suprarrenales/complicaciones , Neoplasias de las Glándulas Suprarrenales/patología , Neoplasias de las Glándulas Suprarrenales/orina , Adenoma Corticosuprarrenal/complicaciones , Adenoma Corticosuprarrenal/patología , Adenoma Corticosuprarrenal/orina , Adulto , Androstenodioles/orina , Cortodoxona/análogos & derivados , Cortodoxona/orina , Síndrome de Cushing/complicaciones , Síndrome de Cushing/patología , Síndrome de Cushing/orina , Detección Precoz del Cáncer , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Oxidorreductasas/orina , Pregnenodionas/orina , Pregnenos/orina , Pregnenolona/orinaRESUMEN
Context: The classic androgen synthesis pathway proceeds via dehydroepiandrosterone, androstenedione, and testosterone to 5α-dihydrotestosterone. However, 5α-dihydrotestosterone synthesis can also be achieved by an alternative pathway originating from 17α-hydroxyprogesterone (17OHP), which accumulates in congenital adrenal hyperplasia (CAH). Similarly, recent work has highlighted androstenedione-derived 11-oxygenated 19-carbon steroids as active androgens, and in CAH, androstenedione is generated directly from 17OHP. The exact contribution of alternative pathway activity to androgen excess in CAH and its response to glucocorticoid (GC) therapy is unknown. Objective: We sought to quantify classic and alternative pathway-mediated androgen synthesis in CAH, their diurnal variation, and their response to conventional GC therapy and modified-release hydrocortisone. Methods: We used urinary steroid metabolome profiling by gas chromatography-mass spectrometry for 24-hour steroid excretion analysis, studying the impact of conventional GCs (hydrocortisone, prednisolone, and dexamethasone) in 55 adults with CAH and 60 controls. We studied diurnal variation in steroid excretion by comparing 8-hourly collections (23:00-7:00, 7:00-15:00, and 15:00-23:00) in 16 patients with CAH taking conventional GCs and during 6 months of treatment with modified-release hydrocortisone, Chronocort. Results: Patients with CAH taking conventional GCs showed low excretion of classic pathway androgen metabolites but excess excretion of the alternative pathway signature metabolites 3α,5α-17-hydroxypregnanolone and 11ß-hydroxyandrosterone. Chronocort reduced 17OHP and alternative pathway metabolite excretion to near-normal levels more consistently than other GC preparations. Conclusions: Alternative pathway-mediated androgen synthesis significantly contributes to androgen excess in CAH. Chronocort therapy appears superior to conventional GC therapy in controlling androgen synthesis via alternative pathways through attenuation of their major substrate, 17OHP.
Asunto(s)
Hiperplasia Suprarrenal Congénita/tratamiento farmacológico , Andrógenos/metabolismo , Ritmo Circadiano , Glucocorticoides/administración & dosificación , Hidrocortisona/administración & dosificación , 17-alfa-Hidroxipregnenolona/orina , Adolescente , Hiperplasia Suprarrenal Congénita/metabolismo , Hiperplasia Suprarrenal Congénita/orina , Adulto , Androsterona/análogos & derivados , Androsterona/orina , Cortodoxona/análogos & derivados , Cortodoxona/orina , Preparaciones de Acción Retardada , Dexametasona/uso terapéutico , Femenino , Cromatografía de Gases y Espectrometría de Masas , Glucocorticoides/uso terapéutico , Humanos , Hidrocortisona/uso terapéutico , Masculino , Persona de Mediana Edad , Prednisolona/uso terapéutico , Pregnanotriol/análogos & derivados , Pregnanotriol/orina , Adulto JovenRESUMEN
Urinary steroid profiling (USP) was studied using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) methods in 108 patients with adrenocortical adenoma (ACA) and in 31 patients with adrenocortical carcinoma (ACC). Thirteen ACC and Cushing's syndrome (ACC-CS) patients had two types of USP as well as 18 ACC patients without hypercortisolism. These four types differed by androgen and glucocorticoid secretion of the adrenal cortex. Fifteen main ACC features were observed by GC-MS. Urinary excretion of dehydroepiandrosterone (DHEA) was increased in 67.7 % of ACC patients and tetrahydro-11-deoxycortisol (THS) in 74.2 %. By combination of the following parameters: THS >900 µg/24 h and/or DHEA >1500 µg/24 h with ratios of 3α,16,20-pregnentriol/3ß,16,20-pregnentriol (3α,16,20dP3/3ß,16,20dP3) less than 6.0 and 3α,17,20dP3/3ß,17,20dP3 less than 9.0 and the detection of "non-classical" 5-en-pregnens, not found in ACA and healthy persons, 100 % sensitivity and specificity of ACC and ACA differential diagnosis were achieved. Features of 21-hydroxylase and 11ß-hydroxylase deficiency were observed by GC-MS in 32.2 and 61.3 % of the ACC patients, respectively. Additional features for ACC-CS diagnostic were increased urinary excretion of 6ß-hydroxycortisol, 18-hydroxycorticosterone, the sum (UFF + UFE) obtained by HPLC, tetrahydrocorticosterone, and the sum (THF + THE + allo-THF) obtained by GC-MS.
Asunto(s)
Neoplasias de la Corteza Suprarrenal/orina , Adenoma Corticosuprarrenal/orina , Carcinoma Corticosuprarrenal/orina , Cromatografía Líquida de Alta Presión/métodos , Cromatografía de Gases y Espectrometría de Masas/métodos , Esteroides/orina , Neoplasias de la Corteza Suprarrenal/diagnóstico , Adenoma Corticosuprarrenal/diagnóstico , Carcinoma Corticosuprarrenal/diagnóstico , Adulto , Cortodoxona/análogos & derivados , Cortodoxona/orina , Síndrome de Cushing/orina , Deshidroepiandrosterona/orina , Diagnóstico Diferencial , Femenino , Humanos , Masculino , Persona de Mediana Edad , Esteroide 11-beta-Hidroxilasa/metabolismo , Esteroide 21-Hidroxilasa/metabolismo , Adulto JovenRESUMEN
Radiological examination may unexpectedly reveal an adrenal mass. Current algorithms for differentiating between benign and malignant lesions mainly rely on size and densitometry on unenhanced CT, which have limited specificity. We examined the diagnostic value of urinary steroid profiling by gas chromatography/mass-spectrometry (GC/MS) in differentiating between benign and malignant adrenal tumors. A retrospective study in two referral centers for patients with adrenal disease was performed. All urinary steroid profiles ordered for evaluation of an adrenal tumor between January 2000 and November 2011 were examined. Patients were diagnosed with adrenal cortical carcinoma (ACC), adrenal cortical adenoma (ACA), or other adrenal mass. Results of hormonal measurements, imaging studies, pathology reports, and clinical outcome were retrieved from medical records. The diagnostic value of individual urinary steroid metabolites was determined by receiver operating characteristics analysis. Cut-off values were compared to reference values from an age and gender-standardized population of healthy controls. Eighteen steroid metabolites were excreted in significantly higher concentrations in patients with ACC (n = 27) compared to patients with ACA (n = 107) or other adrenal conditions (n = 18). Tetrahydro-11-deoxycortisol (THS) at a cut-off value of 2.35 µmol/24 h differentiated ACC from other adrenal disorders with 100% sensitivity and 99% specificity. Elevated urinary excretion of THS was associated with a very high sensitivity and specificity to differentiate between an ACC and a benign adrenal mass. Urinary steroid profiling might be a useful diagnostic test for the evaluation of patients with an adrenal incidentaloma.
Asunto(s)
Neoplasias de la Corteza Suprarrenal/diagnóstico , Carcinoma/diagnóstico , Esteroides/orina , Adulto , Anciano , Estudios de Cohortes , Cortodoxona/análogos & derivados , Cortodoxona/orina , Femenino , Humanos , Masculino , Persona de Mediana Edad , PronósticoRESUMEN
OBJECTIVE: To characterize the urinary steroid metabolome of neonates and infants born either at term or preterm. STUDY DESIGN: We retrospectively analyzed urinary steroid hormone metabolites determined by gas chromatography-mass spectrometry of 78 neonates and infants born at term and 83 neonates and infants born preterm (median 34 weeks of gestational age). The subjects' 11ß-hydroxylase and 21-hydroxylase activities were assessed on the basis of urinary metabolite substrate-to-product ratios. RESULTS: Preterm neonates and infants had elevated urinary concentrations of 17α-hydroxyprogesterone (17OHP) metabolites (P<.001) but lower urinary concentrations of the 21-deoxycortisol metabolite pregnanetriolone (PTO) (P<.01). One reason was lower 11ß-hydroxylase activity in preterms. We could demonstrate a correlation between low 11ß-hydroxylase activity and high urinary concentrations of 17OHP metabolites (r=0.51, P<.001) but low urinary concentrations of the 21-deoxycortisol metabolite PTO (r=-0.24, P=.03) in preterms. CONCLUSIONS: Low 11ß-hydroxylase activity may explain increased 17OHP but decreased 21-deoxycortisol metabolite excretion in preterms. Our analysis clarifies, first, why preterms have higher 17OHP levels and thus higher rates of false-positive screening results for congenital adrenal hyperplasia than do term infants, and, second, why 21-deoxycortisol or its urinary metabolite PTO is more specific than 17OHP for the diagnosis of 21-hydroxylase deficiency.
Asunto(s)
17-alfa-Hidroxiprogesterona/orina , Hiperplasia Suprarrenal Congénita/enzimología , Hiperplasia Suprarrenal Congénita/orina , Recien Nacido Prematuro , Esteroide 11-beta-Hidroxilasa/sangre , Cromatografía de Gases , Cortodoxona/orina , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Lactante , Recién Nacido , Masculino , Espectrometría de Masas , Metaboloma , Pregnanotriol/análogos & derivados , Pregnanotriol/orina , Estudios Retrospectivos , Esteroide 17-alfa-Hidroxilasa/sangreRESUMEN
Adrenocortical carcinoma (ACC) is a very rare malignant tumor with poor prognosis. To gain insight into the pathogenic significance of ACC, we studied clinicopathological features and gene expression profile in ACC. We analyzed five ACC cases (two men and three women) with the median age of 45-year-old who underwent adrenalectomy at our institute. Endocrine studies revealed that two cases had subclinical Cushing's syndrome (SCS) and one with concomitant estrogen-secreting tumor, while the rest of three cases had non-functioning tumors. Analysis of urinary steroids profile by gas chromatography/mass spectrometry showed increased metabolites of corticosteroid precursors, such as 17-OH pregnenolone, 17-OH progesterone, dehydroepiandorosterone (DHEA), and 11-deoxycortisol in all five cases. The pathological diagnosis of ACC was based on Weiss's criteria with its score ≥ 3. The mean size of the resected tumors was 87 mm and Ki67/MIB1 labeling index, a proliferative marker, was 3-27%. Immunohistochemical analysis revealed a disorganized expression of several steroidogenic enzymes, such as 3ß-hydroxysteroid dehydrogenase, 17α-hydroxylase, and DHEA-sulfotransferase. Among several genes determined by RT-PCR, insulin-like growth factor (IGF)-II mRNA was consistently and abundantly expressed in all 5 tumor tissues. Postoperatively, two cases with SCS developed local recurrence and liver metastasis. The present study suggests that the disorganized expression of steroidogenic enzymes and the overexpression of IGF-II by the tumor are hallmarks of ACC, which could be used as biochemical and molecular markers for ACC.
Asunto(s)
17-alfa-Hidroxipregnenolona/análogos & derivados , 17-alfa-Hidroxiprogesterona/metabolismo , Neoplasias de la Corteza Suprarrenal/patología , Carcinoma Corticosuprarrenal/patología , Cortodoxona/metabolismo , Deshidroepiandrosterona/metabolismo , 17-alfa-Hidroxipregnenolona/metabolismo , 17-alfa-Hidroxipregnenolona/orina , 17-alfa-Hidroxiprogesterona/orina , Neoplasias de la Corteza Suprarrenal/metabolismo , Neoplasias de la Corteza Suprarrenal/cirugía , Neoplasias de la Corteza Suprarrenal/orina , Carcinoma Corticosuprarrenal/metabolismo , Carcinoma Corticosuprarrenal/cirugía , Carcinoma Corticosuprarrenal/orina , Adulto , Cortodoxona/orina , Deshidroepiandrosterona/orina , Femenino , Cromatografía de Gases y Espectrometría de Masas , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , ARN Neoplásico/química , ARN Neoplásico/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
We describe an ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC MS/MS) method suitable for a routine laboratory to determine endogenous and exogenous glucocorticoids in plasma, plasma ultrafiltrate, urine and saliva in a single analytical run. After addition of a multi-analyte internal standard, a standardised sample preparation procedure with solid phase extraction followed, before injecting into a tandem mass spectrometer with positive mode electron spray ionisation and multiple reactant monitoring acquisition. The chromatography time was 3min. The limit of quantitation for cortisol and cortisone in plasma was 3.75nmol/L and linearity extended to 2000nmol/L. The limit of quantitation for cortisol in plasma ultrafiltrate and saliva was 0.6nmol/L. The limit of quantitation for 11-deoxycortisol and prednisolone was 5nmol/L and for dexamethasone 1nmol/L. The intra-assay CV was <5% and the inter-assay CV <10% for all analytes in all matrices. Comparison with an immuno-assay (IA) plasma cortisol method resulted in a regression equation of UHPLC=0.79×IA+31.12 with R(2)=0.960 (p<0.0001). Comparison with a high performance liquid chromatography (HPLC) cortisol method yielded a regression equation of UHPLC=1.06×HPLC+9.82, R(2)=0.992 (p<0.0001). The simultaneous measurement of endogenous and exogenous glucocorticoids contributed to patient care in cases with dexamethasone and metyrapone dynamic tests and unsuspected therapeutic glucocorticoid use.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Dexametasona/análisis , Prednisolona/análisis , Pregnenos/análisis , Saliva/química , Espectrometría de Masas en Tándem/métodos , Adulto , Anciano , Cortodoxona/antagonistas & inhibidores , Cortodoxona/sangre , Cortodoxona/orina , Dexametasona/sangre , Dexametasona/orina , Femenino , Humanos , Hidrocortisona/análisis , Hidrocortisona/sangre , Hidrocortisona/orina , Modelos Lineales , Masculino , Persona de Mediana Edad , Prednisolona/sangre , Prednisolona/orina , Pregnenos/sangre , Pregnenos/orina , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Adulto JovenRESUMEN
We use gas chromatography-mass spectrometry (GC-MS) to determine the urine peak area ratio of tetrahydrocortisol (THF) to tetrahydrodeoxycortisol (THS) in spot urine samples of eight male volunteers after a single intramuscular injection of 100 mg hydrocortisone (HC) and after a single oral administration of 10 mg HC at six different post-treatment times over 24 h with 1 week between the two treatments. Control spot urine samples were also obtained from a group of 100 volunteers of each sex for GC-MS analysis. In addition, one female volunteer was collected for GC-MS and isotope ratio mass spectrometry (IRMS) analysis after a single oral administration of 40 mg HC and 40 mg cortisone (C) at 15 and 10 different post-treatment times over 30 h, respectively. IRMS analysis focused on the acetylated derivative of 11-keto-etiocholanolone (11KE) and 11beta-hydroxy-etiocholanolone (11OHE) as target metabolites, and on androsterone (A) as an endogenous reference compound (ERC) for calculating the corresponding delta(13)C (per thousand) depletion values. There was a small but significant sex-related difference for the THF/THS ratio in the control group with mean THF/THS ratio values of 10 and 13.5 for women and men, respectively. A cut-off value of 28 (mean+2 S.D.) for the THF/THS ratio offered a narrow detection window with 39% of suspicious samples after HC-oral treatment, and a wide detection window with 94% of suspicious samples after HC-intramuscular administration in men. For the woman the same cut-off value offered a wide detection window after HC and C administration with 100% and 90% of suspicious samples, respectively. On the basis of a cut-off value of 3 per thousand for the delta(13)C (per thousand) depletion, the exogenous origin was widely evidenced for at least one target compound in 93% and 80% of the HC and C samples, respectively. We conclude by discussing the predictive ability of the urine THF/THS ratio and its usefulness in pointing out suspicious samples resulting from the systemic administration of HC and C.
Asunto(s)
Cortisona/administración & dosificación , Cortodoxona/análogos & derivados , Doping en los Deportes , Hidrocortisona/administración & dosificación , Detección de Abuso de Sustancias/métodos , Tetrahidrocortisol/orina , Administración Oral , Antiinflamatorios/administración & dosificación , Antiinflamatorios/química , Estudios de Casos y Controles , Cortisona/química , Cortodoxona/orina , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Hidrocortisona/química , Inyecciones Intramusculares , Masculino , Factores SexualesRESUMEN
CONTEXT: Variation in the region of chromosome 8 including the genes steroid 11beta-hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) influences mineralocorticoid and glucocorticoid metabolism. However, the relative importance of polymorphisms in CYP11B1 and CYP11B2 in determining these phenotypes is unknown. OBJECTIVE: Our objective was to investigate genetic influences of the CYP11B1 and CYP11B2 genes on mineralocorticoid metabolism. DESIGN: We measured 24-h urinary excretion of the key metabolites of the principal mineralocorticoids, glucocorticoids and androgens secreted by the adrenal cortex. We genotyped polymorphisms spanning the CYP11B1 and CYP11B2 genes, which together capture all common variations at the locus. PARTICIPANTS: Participants included 573 members of 105 British Caucasian families ascertained on a hypertensive proband. MAIN OUTCOME MEASURES: We assessed heritability of urinary tetrahydroaldosterone (THAldo) excretion and association of THAldo excretion with genotype. RESULTS: The heritability of THAldo excretion was 52% (P < 10(-6)). There was significant association between THAldo and genotype at several of the CYP11B1/B2 polymorphisms. The strongest association was observed at the rs6387 (2803A/G) polymorphism in intron 3 of CYP11B1 (P = 0.0004). Association followed a codominant model with a 21% higher THAldo excretion per G allele. Genotype at rs6387 accounted for 2.1% of the total population variability of THAldo. We found significant association between THAldo excretion and urinary total androgen excretion, urinary tetrahydrodeoxycortisol level, and urinary cortisol metabolites (all P < 0.001). CONCLUSIONS: Aldosterone synthesis is highly heritable and is affected by genotype at CYP11B1. Our findings support the hypothesis that genetically determined differences in 11-hydroxylation efficiency can have downstream effects on mineralocorticoid synthesis. Such effects may be of relevance to the development of low-renin essential hypertension.
Asunto(s)
Aldosterona/biosíntesis , Variación Genética , Esteroide 11-beta-Hidroxilasa/genética , Aldosterona/análogos & derivados , Aldosterona/metabolismo , Aldosterona/orina , Cortodoxona/análogos & derivados , Cortodoxona/orina , Citocromo P-450 CYP11B2/genética , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Esteroide 11-beta-Hidroxilasa/metabolismo , Esteroides/orinaRESUMEN
OBJECTIVE: To further analyse the significance and mutual relationship of thyroid function-linked alterations in cortisol metabolism that have been separately and variously reported. PATIENTS AND MEASUREMENTS: Twenty-four-hour urine samples from 21 patients with hyperthyroidism (Graves' disease), 16 patients with hypothyroidism (Hashimoto's thyroiditis), 21 healthy age- and sex-matched controls for hyperthyroidism, and 16 healthy age- and sex-matched controls for hypothyroidism were evaluated for 6beta-hydroxycortisol (6beta-OHF), tetrahydrocortisol (THF), tetrahydrocortisone (THE), allo-tetrahydrocortisol (allo-THF), urinary free cortisol (UFF), urinary free cortisone (UFE) and 17-hydroxycorticosteroid (17-OHCS). RESULTS: Urinary 17-OHCS, THE and allo-THF levels increased considerably in hyperthyroid patients compared to the controls, while UFF and THF showed no difference between the two groups. Urinary 6beta-OHF was significantly lower in the hyperthyroid patients than in the controls. Both the urinary allo-THF + THF/THE and the UFF/UFE ratios were significantly lower in the hyperthyroid patients than in the controls, whereas only the former was significantly higher in the hypothyroid patients than in the controls. The urinary allo-THF/THF ratio was significantly higher in the hyperthyroid patients and significantly lower in the hypothyroid patients than in the controls. In an analysis of pooled subjects including all groups (n = 64), free T4 levels correlated negatively (P < 0.0001) with the urinary allo-THF + THF/THE ratio but not with the UFF/UFE ratio. The serum levels of free T4 correlated positively (P < 0.0001) with the urinary allo-THF/THF ratio. CONCLUSION: The thyroid hormones seem to affect the total 11beta-HSD activity (allo-THF + THF/THE) more strongly than the renal 11beta-HSD2 activity (UFF/UFE). 5alpha-reductase activity (allo-THF/THF) is also enhanced in hyperthyroidism, while the reduction of urinary 6beta-OHF in hyperthyroidism might be a secondary effect of the altered activity of the total 11beta-HSD and 5alpha-reductase.
Asunto(s)
Enfermedad de Graves/orina , Enfermedad de Hashimoto/orina , Hidrocortisona/metabolismo , 17-Hidroxicorticoesteroides/orina , Adulto , Anciano , Estudios de Casos y Controles , Cortisona/orina , Cortodoxona/análogos & derivados , Cortodoxona/orina , Femenino , Humanos , Hidrocortisona/análogos & derivados , Hidrocortisona/orina , Análisis de los Mínimos Cuadrados , Masculino , Persona de Mediana Edad , Tetrahidrocortisol/orina , Tetrahidrocortisona/orinaAsunto(s)
Cortodoxona/análogos & derivados , Enfermedad de Addison/diagnóstico , Enfermedades de las Glándulas Suprarrenales/diagnóstico , Hormona Adrenocorticotrópica , Biomarcadores/orina , Cromatografía en Papel , Colorimetría , Cortodoxona/orina , Síndrome de Cushing/diagnóstico , Dexametasona , Cromatografía de Gases y Espectrometría de Masas , Humanos , Metirapona , Radioinmunoensayo , Valores de Referencia , Manejo de Especímenes , Enfermedades de la Tiroides/diagnósticoRESUMEN
AIMS: Left ventricular mass (LVM) is under the control of aldosterone and angiotensin II in experimental hypertension, but the effect of aldosterone on LVM is controversial in essential hypertension (EH). Some EH patients show a mild impairment of 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) activity without clinical features of the syndrome of apparent mineralocorticoid excess, where the incomplete cortisol-to-cortisone conversion leads to glucocorticoid-mediated mineralocorticoid effects. The mineralocorticoid receptor and 11beta-HSD2 are co-expressed in human heart. We investigated whether LVM may be regulated by glucocorticoids in EH patients. METHODS AND RESULTS: The ratio between 24 h urinary tetrahydro derivatives of cortisol and cortisone (THFs/THE), plasma renin activity, 24 h urinary aldosterone, blood pressure, and LVM indexed for height(2.7) (LVMh(2.7)) were analysed in 493 never-treated hypertensives and 98 normotensives. THFs/THE was associated with LVMh(2.7) in hypertensives and normotensives (r=0.32, P<0.001, and r=0.17, P=0.04, respectively) and persisted after adjusting for confounders (multiple regression analysis). Body mass index, sex, recumbent plasma renin activity, and THFs/THE accounted for 26.1% of LVMh(2.7) variation. Urinary aldosterone was not correlated with LVMh(2.7). CONCLUSION: We suggest that glucocorticoids may take part in the regulation of LVM in EH patients as a function of 11beta-HSD2 activity, and contribute to the target organ damage associated with essential hypertension.
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11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 2/metabolismo , Cortodoxona/análogos & derivados , Glucocorticoides/metabolismo , Hipertensión/enzimología , Hipertrofia Ventricular Izquierda/enzimología , Aldosterona/orina , Presión Sanguínea/fisiología , Estudios de Cohortes , Cortodoxona/orina , Ecocardiografía/métodos , Femenino , Humanos , Hipertensión/patología , Hipertensión/orina , Hipertrofia Ventricular Izquierda/patología , Hipertrofia Ventricular Izquierda/orina , Masculino , Persona de Mediana Edad , Renina/metabolismo , Tetrahidrocortisol/orina , Tetrahidrocortisona/orinaRESUMEN
Genetic variation in the gene encoding aldosterone synthase (CYP11B2) has previously been shown to be associated with hypertension and left ventricular hypertrophy. The intermediate phenotype most consistently associated with variation at this locus is that of elevated plasma 11-deoxycortisol (S). However, in normal subjects, aldosterone synthase does not metabolize S, which is converted to cortisol (F) by the enzyme 11 beta hydroxylase, encoded by the gene CYP11B1, which lies adjacent to CYP11B2 on chromosome 8. It is possible that the quantitative trait locus for the phenotype is within CYP11B1 and that linkage disequilibrium across the extended locus could account for these observations. However, variation across the whole CYP11B1/B2 locus had not been extensively characterized with respect to these phenotypes. We genotyped six polymorphisms in the CYP11B2 gene and three polymorphisms in the CYP11B1 gene in 248 Caucasian nuclear families comprising 1428 individuals. We measured plasma levels of S and F in 460 individuals from 86 families and urinary excretion rates of tetrahydrodeoxycortisol (THS) and tetrahydrodeoxycorticosterone in 573 individuals from 105 families. We examined heritability of the phenotypes and their association with genotypes and haplotypes at this locus. All steroid phenotypes except urinary tetrahydrodeoxycorticosterone were highly heritable (P < 0.00001). There was strong linkage disequilibrium across the CYP11B1/B2 locus. There was modest evidence for association between polymorphisms of CYP11B2 and plasma levels of S (P = 0.02 for T4986C polymorphism) and the plasma S to F ratio, reflecting the activity of 11-beta hydroxylase (P = 0.01 for T4986C polymorphism). There was strong evidence for association between polymorphisms of both CYP11B1 and CYP11B2 and urinary THS, which was strongest for the CYP11B1 exon 1 polymorphism (P = 0.00002). Addition of other marker data to CYP11B1 exon 1 did not improve the fit of a log-linear model. Genotype at CYP11B1 explained approximately 5% of the variance in urinary THS excretion in the population. Thus, it is likely that linkage disequilibrium between causative CYP11B1 variants and CYP11B2 polymorphisms account for the previous observations. Further fine-mapping studies across the CYP11B1 locus are required to localize the causative variant(s) for the biochemical phenotype; this may also identify susceptibility alleles for hypertension and left ventricular hypertrophy.
Asunto(s)
Cortodoxona/orina , Citocromo P-450 CYP11B2/genética , Variación Genética , Desequilibrio de Ligamiento/genética , Polimorfismo Genético , Esteroide 11-beta-Hidroxilasa/genética , Corticoesteroides/sangre , Presión Sanguínea , Mapeo Cromosómico , Exones/genética , Femenino , Genotipo , Humanos , Intrones/genética , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple/genética , Caracteres Sexuales , Reino Unido , Población Blanca/genéticaRESUMEN
OBJECTIVE: Aldosterone synthase, a key enzyme in the terminal steps of aldosterone synthesis, is encoded by the CYP11B2 gene. A polymorphism in the 5' coding region of this gene (-344 C/T) is associated with hypertension, particularly with elevation of the aldosterone to renin ratio. A second polymorphism (a conversion in intron 2 to resemble that of the neighbouring 11beta-hydroxylase (CYP11B1) gene) is found in close linkage dysequilibrium with the variant at -344 C/T. The mechanism by which these variants predispose to cardiovascular disease and the precise intermediate phenotype associated with them remains speculative. DESIGN: We performed a focused physiological study in normal volunteers stratified by CYP11B2 genotype. PATIENTS: Twenty-three subjects homozygous for the T allele and 21 homozygous for the C allele of the -344 C/T polymorphism of CYP11B2 were studied. MEASUREMENTS: Basal and angiotensin II stimulated plasma and 24-h urinary steroid excretion during low (60 mmol/day) and high (160 mmol/day) sodium intake and plasma steroids after ACTH stimulation were measured. RESULTS: No influence of polymorphic variation on basal or stimulated plasma cortisol or aldosterone or other plasma steroid concentrations during either dietary phase was seen. However, excretion of tetrahydro-11-deoxycortisol (the urinary metabolite of 11-deoxycortisol), which is the precursor of cortisol) was increased in TT subjects during sodium restriction, consistent with impairment of zona fasciculata 11beta-hydroxylation. CONCLUSIONS: We conclude that this polymorphism has no major influence on normal zona glomerulosa function but is associated with a change in 11beta-hydroxylation in the zona fasciculata. The mechanism remains uncertain, but alteration of 11-deoxycortisol levels without change in cortisol suggests altered efficiency of 11beta-hydroxylation. In the long term, this may lead to a minor but chronic increase in ACTH drive to the gland, which may have consequences for steroid synthesis and predispose to the risk of cardiovascular disease.
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Corteza Suprarrenal/fisiología , Hormona Adrenocorticotrópica/administración & dosificación , Angiotensina II/administración & dosificación , Cortodoxona/análogos & derivados , Citocromo P-450 CYP11B2/genética , Polimorfismo Genético/genética , Sodio en la Dieta/administración & dosificación , Adulto , Aldosterona/sangre , Corticosterona/sangre , Cortodoxona/sangre , Cortodoxona/metabolismo , Cortodoxona/orina , Estudios Cruzados , Desoxicorticosterona/sangre , Método Doble Ciego , Homocigoto , Humanos , Hidrocortisona/sangre , Hidrocortisona/orina , Infusiones Intravenosas , MasculinoRESUMEN
The terminal stages in the synthesis of aldosterone and cortisol are catalysed by the enzymes aldosterone synthase and 11beta-hydroxylase respectively. We have previously reported that polymorphic variation in the 5' promoter region (-344C/T) of the gene encoding aldosterone synthase (CYP11B2) is associated with increased aldosterone metabolite excretion and with hypertension associated with a raised aldosterone to renin ratio (ARR). Additionally, basal and ACTH-stimulated plasma levels of 11-deoxycortisol, the precursor of cortisol, are higher in subjects carrying the T-allelic variant. We have now identified in a family study (573 individuals from 105 extended families ascertained through a hypertensive proband) that excretion of the main metabolite of this steroid (tetrahydro-11-deoxycortisol, THS) is heritable (19.4%) and that the T-allele of CYP11B2 is more strongly associated with higher THS levels than the C-allele. Raised plasma and urinary levels of 11-deoxycortisol suggest that there is relative inefficiency of 11beta-hydroxylation in the zona fasciculata; the P450 enzyme responsible for this step is encoded by the gene CYP11B1, which is highly homologous with and adjacent to CYP11B2. The association of genetic variation in the promoter of CYP11B2 which, in the adrenal cortex, is only expressed in zona glomerulosa, and zona fasciculata 11beta-hydroxylation function is paradoxical. There may be linkage dys-equilibrium between this polymorphism and a quantitative trait locus (QTL) in CYP11B1. Chronic alteration of 11beta-hydroxylase activity may increase ACTH drive to the adrenal cortex, altering the regulation of aldosterone synthesis. This may explain, at least partly, the association between CYP11B2 polymorphisms and hypertension.
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Aldosterona/biosíntesis , Presión Sanguínea/genética , Citocromo P-450 CYP11B2/genética , Hipertensión/genética , Mutación Puntual , Regiones Promotoras Genéticas/genética , Hormona Adrenocorticotrópica/metabolismo , Aldosterona/genética , Presión Sanguínea/fisiología , Cortodoxona/sangre , Cortodoxona/orina , Citocromo P-450 CYP11B2/metabolismo , Predisposición Genética a la Enfermedad , Haplotipos , Humanos , Hidrocortisona/metabolismo , Hipertensión/sangre , Hipertensión/fisiopatología , Hipertensión/orina , Polimorfismo Genético , Regiones Promotoras Genéticas/fisiología , Sitios de Carácter Cuantitativo/genética , Esteroide 11-beta-Hidroxilasa/metabolismo , Zona Fascicular/fisiopatologíaRESUMEN
In glucocorticoid-remediable aldosteronism (GRA), there is a large interfamily variation of phenotype. We report three subjects with GRA in a single family (parents, two brothers and two sisters), of whom only one (proband) displayed classical features of the mineralocorticoid excess. The proband was a man found to be hypertensive and hypokalaemic at the age of 24 years. Plasma renin activity was suppressed and plasma aldosterone was repeatedly elevated. Blood pressure and aldosterone levels normalized within 5 days of dexamethasone therapy. The presence of a chimaeric CYP11B1/CYP11B2 gene was demonstrated by long-PCR and Southern blotting (crossover site at the end of intron 3) in the proband, in the younger sister (sibling 1) and in the father. In these patients, sequencing of the chimaeric portion of CYP11B1 did not reveal any mutation, while sequencing of the chimaeric portion of CYP11B2 showed a V386A polymorphism in exon 7, known to cause only a minimal impairment of enzymatic activity. Sibling 1 was normotensive, normokalaemic and had normal PRA and aldosterone. The father had normal blood pressure and potassium, low-normal PRA and normal aldosterone. All three subjects had elevated levels of urinary 18-hydroxycortisol and 18-oxocortisol. Baseline 11-deoxycorticosterone (DOC), corticosterone (B) and aldosterone were high in the proband and normal in the father and sibling 1; 11-deoxycortisol (S) and cortisol (F) were normal. ACTH induced a normal increase of B, DOC, S and F, and an excessive aldosterone increase in all three patients. Abnormalities in the chimaeric portions of CYB11B1 or CYP11B2 genes did not account for the phenotypic disparity of the different members in a single GRA family. Altered regulation of the chimaeric gene may be responsible for differences in its activity.
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Dexametasona/uso terapéutico , Glucocorticoides/uso terapéutico , Hidrocortisona/análogos & derivados , Hiperaldosteronismo/genética , Adulto , Anciano , Aldosterona/sangre , Aldosterona/orina , Cortodoxona/orina , Citocromo P-450 CYP11B2/genética , Femenino , Genotipo , Humanos , Hidrocortisona/orina , Hiperaldosteronismo/tratamiento farmacológico , Hiperaldosteronismo/metabolismo , Hipertensión/genética , Masculino , Persona de Mediana Edad , Linaje , Fenotipo , Renina/sangre , Renina/orina , Esteroide 11-beta-Hidroxilasa/genéticaRESUMEN
OBJECTIVES: To study the effect of endogenous steroids on the presence of uterine leiomyomas. METHODS: Urine samples of 27 premenopausal women with leiomyomas and 25 age-matched healthy premenopausal women were collected. The concentration of estrogens and androgens in the urine samples of the two groups were determined using a gas chromatography mass spectrometer and the two groups were compared. To study metabolic changes in patients indirectly, the concentration ratios of precursor metabolite to product metabolite of the two groups were also compared. RESULTS: Urinary concentrations of 17beta-estradiol, 5-androstene-3beta, 16beta, 17beta, triol, 11-keto-ethiocholanolone, 11beta-hydroxy-androsterone, 11beta-hydroxy-etiocholanolone, THS, THA, THE, alpha-cortol and beta-cortol were significantly higher in patients than in controls. The concentration ratios of 17beta-estradiol/estrone and 11/beta-hydroxy-ethiocholanolone/11beta-hydroxy-androsterone increased in patients. CONCLUSIONS: The presence of uterine leiomyomas correlates with an increase in urinary concentrations of estrogens and androgens, and it appears to be caused by a decrease in patients' metabolism of steroids.
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Andrógenos/orina , Androsterona/análogos & derivados , Corticosterona/análogos & derivados , Cortodoxona/análogos & derivados , Estrógenos/orina , Etiocolanolona/análogos & derivados , Leiomioma/metabolismo , Neoplasias Uterinas/metabolismo , Adulto , Androstenoles/orina , Androsterona/orina , Estudios de Casos y Controles , Corticosterona/orina , Cortodoxona/orina , Deshidroepiandrosterona/orina , Estradiol/orina , Estrona/orina , Etiocolanolona/orina , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Leiomioma/orina , Persona de Mediana Edad , Pregnanos/orina , Premenopausia , Tetrahidrocortisona/orina , Neoplasias Uterinas/orinaRESUMEN
BACKGROUND: It has been suggested that an altered setpoint of the 11betaHSD-mediated cortisol to cortisone interconversion towards cortisol contributes to sodium retention in nephrotic syndrome patients. We studied the parameters of 11betaHSD activity in proteinuric patients, in particular its activity at the kidney level. We also studied the effect of angiotensin-II receptor blockade on the parameters of 11betaHSD activity. MATERIALS AND METHODS: Serum cortisol/cortisone ratio and the urinary ratios of (tetrahydrocortisol + allo-tetrahydrocortisol)/tetrahydrocortisone [(THF + allo-THF)/THE] and of urinary free cortisol/free cortisone (UFF/UFE) were measured in eight proteinuric patients and compared with eight matched, healthy subjects. Patients were subsequently studied after 4 weeks' treatment with losartan 50 mg day-1 and placebo, respectively. RESULTS: No significant differences between the proteinuric patients and the healthy subjects were observed in the serum cortisol, serum cortisone, serum cortisol to cortisone ratio, or in the urinary excretions of THF, allo-THF, THE, sum of cortisol metabolites, or the (THF + allo-THF)/THE ratio. Urinary free cortisol excretion and the UFF/UFE ratio were lower in the proteinuric patients than in the healthy subjects (56 +/- 21 vs. 85 +/- 24 pmol min-1, P < 0.05, and 0.39 +/- 0.07 vs. 0.63 +/- 0.28, P < 0.05, respectively). Mean arterial pressure and proteinuria were reduced significantly during losartan treatment, but without concomitant changes in peripheral cortisol metabolism. CONCLUSIONS: Increased renal inactivation of cortisol in proteinuric patients does not support the contention that altered 11betaHSD activity contributes to sodium retention in patients with nephrotic syndrome. Losartan 50 mg d.d. reduces mean arterial pressure and proteinuria, but does not exert a significant effect on the cortisol to cortisone interconversion.
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Antagonistas de Receptores de Angiotensina , Cortodoxona/análogos & derivados , Hidroxiesteroide Deshidrogenasas/metabolismo , Riñón/enzimología , Losartán/uso terapéutico , Proteinuria/tratamiento farmacológico , Proteinuria/enzimología , 11-beta-Hidroxiesteroide Deshidrogenasas , Adulto , Estudios de Casos y Controles , Cortisona/sangre , Cortisona/orina , Cortodoxona/orina , Femenino , Humanos , Hidrocortisona/sangre , Hidrocortisona/orina , Masculino , Proteinuria/metabolismo , Tetrahidrocortisol/orina , Tetrahidrocortisona/orinaRESUMEN
Relative 11beta-hydroxysteroid dehydrogenase deficiency has been shown previously to arise from endogenous hypercortisolism in diseases of the hypothalamic/pituitary/adrenocortical system; whether stress induced hypercortisolism may also result in substrate overload of 11beta-hydroxysteroid dehydrogenase has not yet been studied. We therefore studied the characteristics of cortisol metabolisation during the postoperative period of cardiac surgery, representing a well standardized surgical procedure. In a prospective, observational, consecutive case study, 14 patients undergoing cardiac surgery were investigated. During the first two days after cardiac surgery urine was collected from the patients during two 10 hour overnight periods (8 p.m. (day of surgery) until 6 a.m., and during the following night). Using capillary gas-chromatography, main urinary cortisol metabolites were quantified (tetrahydrocortisone, tetrahydrocortisol, allo-tetrahydrocortisol, cortolones, cortols as sum of cortisol metabolites (CM)). Free urinary cortisol (FUC) was determined by an automated immunoassay after extraction. The ratio of cortisol metabolites (tetrahydrocortisol, allo-tetrahydrocortisol, cortols) to cortisone metabolites (tetrahydrocortisone, cortolones) was calculated to characterize the overall activity of 11beta-hydroxysteroid dehydrogenase, an enzyme system catalyzing the conversion of cortisol to inactive cortisone (CMR, cortisol metabolisation ratio). Total cortisol metabolisation (including hepatic ring A-reduction and conjugation) was estimated by a cortisol turnover quotient (CM/FUC). In all urinary samples the ratio of cortisol to cortisone metabolites was markedly elevated compared to controls (patients: median 1.9, interquartile range 1.5-2.4, absolute range 1.0-3.2; controls: median 0.45, interquartile range 0.36-0.52); this ratio was positively correlated to FUC (r2 = 0.30; p = 0.003). The cortisol turnover quotient was markedly reduced (patients: median 38.0, interquartile range 20.0-103.9, absolute range 8.3-211.9; controls: median 259, interquartile range 176-415) and inversely correlated to FUC (r2 = 0.64, p < 0.001). It is concluded that major surgical trauma results in a marked relative reduction of cortisol inactivation probably consequent to substrate overload of the metabolizing enzymes; as the activity of these enzymes (mainly 11beta-hydroxysteroid dehydrogenase) is crucial for the modulation of cortisol receptor access, tissue corticoid sensitivity in the postoperative period may vary substantially from physiological conditions.