RESUMO
CONTEXT: Oral once-daily dual-release hydrocortisone (DR-HC) replacement therapy has demonstrated an improved metabolic profile compared to conventional 3-times-daily (TID-HC) therapy among patients with primary adrenal insufficiency. This effect might be related to a more physiological cortisol profile, but also to a modified pattern of cortisol metabolism. OBJECTIVE: This work aimed to study cortisol metabolism during DR-HC and TID-HC. DESIGN: A randomized, 12-week, crossover study was conducted. INTERVENTION AND PARTICIPANTS: DC-HC and same daily dose of TID-HC were administered to patients with primary adrenal insufficiency (nâ =â 50) vs healthy individuals (nâ =â 124) as controls. MAIN OUTCOME MEASURES: Urinary corticosteroid metabolites were measured by gas chromatography/mass spectrometry at 24-hour urinary collections. RESULTS: Total cortisol metabolites decreased during DR-HC compared to TID-HC (Pâ <â .001) and reached control values (Pâ =â .089). During DR-HC, 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) activity measured by tetrahydrocortisolâ +â 5α-tetrahydrocortisol/tetrahydrocortisone ratio was reduced compared to TID-HC (Pâ <â .05), but remained increased vs controls (Pâ <â .001). 11ß-HSD2 activity measured by urinary free cortisone/free cortisol ratio was decreased with TID-HC vs controls (Pâ <â .01) but normalized with DR-HC (Pâ =â .358). 5α- and 5ß-reduced metabolites were decreased with DR-HC compared to TID-HC. Tetrahydrocortisol/5α-tetrahydrocortisol ratio was increased during both treatments, suggesting increased 5ß-reductase activity. CONCLUSIONS: The urinary cortisol metabolome shows striking abnormalities in patients receiving conventional TID-HC replacement therapy, with increased 11ß-HSD1 activity that may account for the unfavorable metabolic phenotype in primary adrenal insufficiency. Its change toward normalization with DR-HC may mediate beneficial metabolic effects. The urinary cortisol metabolome may serve as a tool to assess optimal cortisol replacement therapy.
Assuntos
Doença de Addison , Hidrocortisona/farmacocinética , Esteroides/urina , Doença de Addison/tratamento farmacológico , Doença de Addison/metabolismo , Doença de Addison/urina , Adulto , Idoso , Cortisona/metabolismo , Cortisona/urina , Estudos Cross-Over , Preparações de Ação Retardada/farmacocinética , Preparações de Ação Retardada/uso terapêutico , Europa (Continente) , Feminino , Humanos , Hidrocortisona/uso terapêutico , Hidrocortisona/urina , Masculino , Metaboloma/efeitos dos fármacos , Pessoa de Meia-Idade , Pregnanos/metabolismo , Pregnanos/urina , Esteroides/metabolismo , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Tetra-Hidrocortisona/urina , UrináliseRESUMO
Successful translation of laboratory-based surface-enhanced Raman scattering (SERS) platforms to clinical applications requires multiplex and ultratrace detection of small biomarker molecules from a complex biofluid. However, these biomarker molecules generally exhibit low Raman scattering cross sections and do not possess specific affinity to plasmonic nanoparticle surfaces, significantly increasing the challenge of detecting them at low concentrations. Herein, we demonstrate a "confine-and-capture" approach for multiplex detection of two families of urine metabolites correlated with miscarriage risks, 5ß-pregnane-3α,20α-diol-3α-glucuronide and tetrahydrocortisone. To enhance SERS signals by 1012-fold, we use specific nanoscale surface chemistry for targeted metabolite capture from a complex urine matrix prior to confining them on a superhydrophobic SERS platform. We then apply chemometrics, including principal component analysis and partial least-squares regression, to convert molecular fingerprint information into quantifiable readouts. The whole screening procedure requires only 30 min, including urine pretreatment, sample drying on the SERS platform, SERS measurements, and chemometric analyses. These readouts correlate well with the pregnancy outcomes in a case-control study of 40 patients presenting threatened miscarriage symptoms.
Assuntos
Pregnanodiol/urina , Tetra-Hidrocortisona/urina , Calibragem , Teoria da Densidade Funcional , Feminino , Humanos , Estrutura Molecular , Tamanho da Partícula , Gravidez , Pregnanodiol/análogos & derivados , Pregnanodiol/metabolismo , Análise Espectral Raman , Propriedades de Superfície , Tetra-Hidrocortisona/metabolismo , Fatores de TempoRESUMO
Low-grade metabolic acidosis (LGMA), as induced by high dietary acid load or sodium chloride (NaCl) intake, has been shown to increase bone and protein catabolism. Underlying mechanisms are not fully understood, but from clinical metabolic acidosis interactions of acid-base balance with glucocorticoid (GC) metabolism are known. We aimed to investigate GC activity/metabolism under alkaline supplementation and NaCl-induced LGMA. Eight young, healthy, normal-weight men participated in two crossover designed interventional studies. In Study A, two 10-day high NaCl diet (32 g/d) periods were conducted, one supplemented with 90 mmol KHCO3/day. In Study B, participants received a high and a low NaCl diet (31 vs. 3 g/day), each for 14 days. During low NaCl, the diet was moderately acidified by replacement of a bicarbonate-rich mineral water (consumed during high NaCl) with a non-alkalizing drinking water. In repeatedly collected 24-h urine samples, potentially bioactive-free GCs (urinary-free cortisol + free cortisone) were analyzed, as well as tetrahydrocortisol (THF), 5α-THF, and tetrahydrocortisone (THE). With supplementation of 90 mmol KHCO3, the marker of total adrenal GC secretion (THF + 5α-THF + THE) dropped (p = 0.047) and potentially bioactive-free GCs were reduced (p = 0.003). In Study B, however, GC secretion and potentially bioactive-free GCs did not exhibit the expected fall with NaCl-reduction as net acid excretion was raised by 30 mEq/d. Diet-induced acidification/alkalization affects GC activity and metabolism, which in case of long-term ingestion of habitually acidifying western diets may constitute an independent risk factor for bone degradation and cardiometabolic diseases.
Assuntos
Acidose/induzido quimicamente , Acidose/metabolismo , Álcalis/farmacologia , Glucocorticoides/metabolismo , Cloreto de Sódio , Equilíbrio Ácido-Base/efeitos dos fármacos , Adulto , Bicarbonatos/farmacologia , Cortisona/urina , Estudos Cross-Over , Dieta , Água Potável , Glucocorticoides/urina , Humanos , Hidrocortisona/urina , Masculino , Compostos de Potássio/farmacologia , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Adulto JovemRESUMO
One major issue of newborn screening programs for 21-hydroxylase deficiency (21OHD) is the high rate of false-positive results, especially in preterm neonates. Urinary steroid metabolite analysis using gas chromatography-mass spectrometry (GC-MS) is suitable as a confirmatory diagnostic tool. The objective of this study was to analyze retrospectively diagnostic metabolite ratios in neonates and infants with and without 21OHD using GC-MS with emphasis on glucocorticoid metabolism, and to develop reference values for the steroid metabolite ratios for the diagnosis of 21OHD. We retrospectively analyzed urinary steroid hormone metabolites determined by GC-MS of 95 untreated neonates and infants with 21OHD (1-148 days), and 261 neonates and infants (100 preterms) without 21OHD (0-217 days). Metabolites of 17α-hydroxyprogesterone showed specificities below 98%, whereas the 21-deoxycortisol metabolite pregnanetriolone clearly separated 21OHD from non-21OHD subjects. The best diagnostic ratio for 21OHD was pregnanetriolone to 6α-hydroxy-tetrahydrocortisone. The lowest value of this ratio in the 21OHD group (0.47) was at least eight times higher than the highest values in the non-21OHD group (0.055). We have given appropriate reference values for steroid metabolite ratios in the largest 21OHD cohort so far described. Consideration of glucocorticoid metabolism, especially the use of typical neonatal 6α-hydroxylates metabolites, leads to improvement of diagnostic metabolite ratios.
Assuntos
Hiperplasia Suprarrenal Congênita/diagnóstico , Hiperplasia Suprarrenal Congênita/urina , Cromatografia Gasosa-Espectrometria de Massas/métodos , Metabolômica/métodos , Hiperplasia Suprarrenal Congênita/metabolismo , Estudos de Coortes , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pregnanotriol/análogos & derivados , Pregnanotriol/metabolismo , Pregnanotriol/urina , Valores de Referência , Esteroides/metabolismo , Esteroides/urina , Tetra-Hidrocortisona/análogos & derivados , Tetra-Hidrocortisona/metabolismo , Tetra-Hidrocortisona/urinaRESUMO
Chronic stress as well as major depressive disorders is associated with cortisol metabolism. Two enzymes modulate cortisol (F) and cortisone (E) interconversion: 11ß-hydroxysteroid dehydrogenase type 1 and type 2 (11ß-HSD1 and 11ß-HSD2). Furthermore, F and E were inactivated by 5α and 5ß reductases to their tetrahydro-metabolites: tetrahydrocortisol (THF), allo-tetrahydrocortisol (5α-THF) and tetrahydrocortisone (THE). To better understand depression a LC-MS/MS method for simultaneous determination of F, E THF, 5α-THF and THE in human urine has been developed and validated. The quantification range was 0.1-160 ng mL(-1) for F and E, and 0.2-160 ng mL(-1) for the tetrahydro-metabolites, with >86.1% recovery for all analytes. The nocturnal urine concentrations of F, E and tetrahydro-metabolites in 12 apparently healthy male adult volunteers and 12 drug-free male patients (age range, 20-50 years) with a diagnosis of depression were analyzed. A series of significant changes in glucocorticoid metabolism can be detected: F/E ratios and (THF+5α-THF)/THE ratios as well as F and THF concentrations were significantly higher in depression patients than in healthy subjects (p<0.05); 5α-THF/F ratios, 5α-THF/THF ratios as well as 5α-THF concentrations were significantly lower in depression patients (p<0.05). The results pointed to the decreased 11ß-HSD2 activity and a dysfunction in the 5α-reductase pathway in depressed patients. This method allows the assessment of 11ß-HSD1/2 and 5α/ß-reductase activities in a single analytical run providing an innovative tool to explain the potential etiology of depression.
Assuntos
Cortisona/química , Cortisona/urina , Transtorno Depressivo Maior/urina , Hidrocortisona/química , Hidrocortisona/urina , Tetra-Hidrocortisona/química , Tetra-Hidrocortisona/urina , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2/metabolismo , Adulto , Estudos de Casos e Controles , Colestenona 5 alfa-Redutase/metabolismo , Cromatografia Líquida/métodos , Cortisona/metabolismo , Transtorno Depressivo Maior/metabolismo , Glucocorticoides/química , Glucocorticoides/metabolismo , Glucocorticoides/urina , Humanos , Hidrocortisona/metabolismo , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas em Tandem/métodos , Tetra-Hidrocortisol/análogos & derivados , Tetra-Hidrocortisol/química , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Adulto JovemRESUMO
Dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis is suggested as a pathophysiological factor in bipolar disorder and schizophrenia. Increased clearance of cortisol was recently indicated as a component in the HPA axis hyperdrive. The aim of the present study was to test the model of increased cortisol metabolism in a new replication sample separately and combined with a previously published sample of bipolar disorder and schizophrenia. Spot urine was sampled from 212 healthy controls (HC) and 221 patients with a schizophrenia spectrum disorder (SCZ, n = 115) and bipolar disorder (BD, n = 106). Of these, a subsample of 169 HC and 155 patients was included in a previous report. Urinary free cortisol, cortisone and their metabolites were measured, and the activities of 5α-reductase, 5ß-reductase and 11ß-HSD were estimated and analyzed for differences between groups. In the new sample, there was increased enzyme activity in SCZ for 5ß-reductase (p = 0.024 vs HC; p = 0.027 vs BD) and 11ß-HSD2 (p = 0.014 vs HC; p = 0.004 vs BD). In the combined sample, there was increased activity in SCZ for 5α-reductase (p < 0.001 vs HC; p = 0.020 vs BD), 5ß-reductase (p < 0.001 vs HC; p = 0.045 vs BD) and 11ß-HSD2 (p < 0.001 vs HC; p = 0.043 vs BD), and in BD for 5ß-reductase (p = 0.002), 11ß-HSD2 (p = 0.039) and 5α-reductase (trend, p = 0.084) (all vs HC). The findings confirm increased systemic cortisol metabolism in BD and SCZ. This is most consistent in SCZ, with BD taking an intermediate position. The design makes it impossible to determine the direction of the effect. However, the findings merit further study of cortisol metabolism as a possible component in the HPA axis dysfunction and pathophysiology of BD and SCZ.
Assuntos
Transtorno Bipolar/metabolismo , Hidrocortisona/metabolismo , Esquizofrenia/metabolismo , Adolescente , Adulto , Análise de Variância , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Escalas de Graduação Psiquiátrica , Tetra-Hidrocortisol/análogos & derivados , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismo , Adulto JovemRESUMO
OBJECTIVE: Increased glucocorticoid metabolite excretion and enhanced expression and activity of 11ß-hydroxysteroid dehydrogenase type 1 in adipose tissue are closely correlated with obesity and its detrimental consequences. Weight loss ameliorates the latter. The aim of this study was to explore whether increased glucocorticoid exposure in obesity is improved with substantial weight loss and thus is a consequence rather than a cause of obesity. DESIGN AND PATIENTS: A prospective cohort study in 31 women. MEASUREMENTS: 11ß-HSD type 1 expression and activity, urinary glucocorticoid metabolite excretion, body composition including regional adipose tissue depots and insulin resistance by HOMA-IR before and 2 years after gastric bypass surgery. RESULTS: After weight loss, excretion of cortisol and cortisone metabolites decreased. Both cortisol and cortisone metabolite excretion correlated with central obesity, where the intraabdominal fat depot showed the strongest association. Cortisol metabolites correlated with 11ß-HSD type 1 activity in abdominal subcutaneous adipose tissue. The ratio of cortisol to cortisone metabolites [(5α-tetrahydrocortisol (5αTHF) + tetrahydrocortisol (THF) + α-cortol)/(tetrahydrocortisone (THE) + α-cortolone)] and the ratio of 5α-THF/THF both decreased after stable weight loss, reflecting a downregulation of the net activities of 11ß-HSD type 1 and 5α-reductase. CONCLUSION: Long-term weight loss in women is not only followed by reduced glucocorticoid production, but also favourably decreases the global and tissue-specific activity of the cortisol-activating enzyme 11 ß-HSD type 1, possibly contributing to the health benefits of bariatric surgery.
Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Hidrocortisona/urina , Obesidade/metabolismo , Obesidade/urina , Redução de Peso/fisiologia , Adulto , Estudos de Coortes , Cortisona/urina , Feminino , Humanos , Pessoa de Meia-Idade , Estudos Prospectivos , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismoRESUMO
Intense physical exercise activates the hypothalamic-pituitary-adrenocortical axis but little is known about changes in glucocorticoid sensitivity at the target cell level. No data are available on the acute effects of exercise on 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 activity, which generates biologically active cortisol from inactive cortisone and is expressed also in skeletal muscle. Fifteen healthy, trained males (age mean +/- SE 28 +/- 1) were assessed on three non-consecutive days: at rest, during an endurance and strength sessions. During each session, between 1000 and 1600 hours, 6-h urine and four salivary samples were collected. Urinary total tetrahydrocortisol (THF) + alloTHF, tetrahydrocortisone (THE), cortisol (F) and cortisone (E) were measured with HPLC-tandem mass spectrometry; urinary-unconjugated F and E were measured by HPLC-UV. Salivary cortisol and interleukin (IL)-6 were measured by RIA and ELISA, respectively. Both endurance and strength exercises caused an increase in (THF + alloTHF)/THE ratio (mean +/- SE 1.90 +/- 0.07 and 1.82 +/- 0.05 vs. 1.63 +/- 0.06, P < 0.01 and P = 0.03, respectively), consistent with increased systemic 11beta-HSD type 1 activity. No relationship was found with age, BMI, VO(2max) maximal power load or perceived exertion. No significant change was apparent in F/E ratio, an index of 11beta-HSD type 2 activity. No effect of exercise on salivary cortisol and IL-6 was observed, whereas a significant effect of sampling time was found. Intense physical exercise acutely increases systemic 11beta-HSD type 1 activity in humans. Such an increase may lead to higher cortisol concentration in target tissues, notably in skeletal muscle where it could contribute to limit exercise-induced muscle inflammatory response.
Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Esforço Físico/fisiologia , Adulto , Cortisona/metabolismo , Cortisona/urina , Exercício Físico/fisiologia , Saúde , Humanos , Hidrocortisona/metabolismo , Hidrocortisona/urina , Interleucina-6/metabolismo , Masculino , Resistência Física/fisiologia , Treinamento Resistido , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Tetra-Hidrocortisona/urina , Regulação para CimaRESUMO
Reduced concentration of serum ionized calcium and increased urinary calcium excretion have been reported in primary aldosteronism and glucocorticoid-treated patients. A reduced activity of the 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta HSD2) results in overstimulation of the mineralocorticoid receptor by cortisol. Whether inhibition of the 11 beta HSD2 by glycyrrhetinic acid (GA) may increase renal calcium excretion is unknown. Serum and urinary electrolyte and creatinine, serum ionized calcium, urinary calcium excretion, and the steroid metabolites (THF+5 alpha THF)/THE as a parameter of 11 beta HSD2 activity were repeatedly measured in 20 healthy subjects during baseline conditions and during 1 wk of 500 mg/d GA. One week of GA induced a maximal increment of 93% in (THF+5 alpha THF)/THE. Ambulatory BP was significantly higher at day 7 of GA than at baseline (126/77 +/- 10/7 versus 115/73 +/- 8/6 mmHg; P < 0.001 for systolic; P < 0.05 for diastolic). During GA administration, serum ionized calcium decreased from 1.26 +/- 0.05 to 1.18 +/- 0.04 mmol/L (P < 0.0001), and absolute urinary calcium excretion was enhanced from 29.2 +/- 3.6 to 31.9 +/- 3.1 micromol/L GFR (P < 0.01). Fractional calcium excretion increased from 2.4 +/- 0.3 to 2.7 +/- 0.3% (P < 0.01) and was negatively correlated to the fractional sodium excretion during GA (R = -0.35; P < 0.001). Moreover, serum potassium correlated positively with serum ionized calcium (R = 0.66; P < 0.0001). Inhibition of 11 beta HSD2 activity is sufficient to significantly increase the fractional excretion of calcium and decrease serum ionized calcium, suggesting decreased tubular reabsorption of this divalent cation under conditions of renal glucocorticoid/mineralocorticoid excess. The likely site of steroid-regulated renal calcium handling appears to be the distal tubule.
Assuntos
Cálcio/metabolismo , Ácido Glicirretínico/farmacologia , Hidroxiesteroide Desidrogenases/metabolismo , Rim/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2 , Adulto , Pressão Sanguínea/efeitos dos fármacos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Potássio/sangue , Estudos Prospectivos , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismoRESUMO
This study describes a new approach using stable isotope methodology in evaluating 11beta-HSD activities in vivo based on urinary excretion of cortisol, cortisone, and their A-ring reduced metabolites. The method involved the measurement of deuterium-labeled cortisol and its deuterium-labeled metabolites by GC/MS simultaneously with endogenous cortisol, cortisone, and their A-ring reduced metabolites after oral administration of deuterium-labeled cortisol to normal human subjects. This stable isotope approach offered unique advantages in assessing the appropriateness of measuring unconjugated and total (unconjugated + conjugated) cortisol, cortisone, and their A-ring reduced metabolites in urine as indices of renal 11beta-HSD2 activity in man. Our results strongly support that the measurement of urinary unconjugated cortisol and cortisone is a significant advance in assessing 11beta-HSD2 activity.
Assuntos
Deutério , Hidrocortisona , Hidroxiesteroide Desidrogenases/metabolismo , Rim/enzimologia , 11-beta-Hidroxiesteroide Desidrogenases , Adulto , Cortisona/metabolismo , Cortisona/urina , Deutério/administração & dosagem , Deutério/metabolismo , Humanos , Hidrocortisona/administração & dosagem , Hidrocortisona/metabolismo , Rim/metabolismo , Masculino , Espectrometria de Massas , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Tetra-Hidrocortisona/urina , Fatores de TempoRESUMO
Oral dehydroepiandrosterone (DHEA) replacement therapy may have a multitude of potential beneficial effects and exerts its action mainly via peripheral bioconversion to androgens (and estrogens). A daily dose of 50-mg DHEA has been shown by us and others to restore low endogenous serum DHEA concentrations to normal youthful levels followed by an increase in circulating androgens and estrogens. As the hepatic first-pass effect may lead to a non physiological metabolism of DHEA after oral ingestion we studied the influence of two single DHEA doses (50 and 100 mg) on the excretion of steroid metabolites in 14 elderly males [age 58.8+/-5.1 years (mean +/- SEM)] with endogenous DHEAS levels <1500 ng/ml and in 9 healthy females (age 23.3+/-4.1 years) with transient suppression of endogenous DHEA secretion induced by dexamethasone (dex) pretreatment (4x0.5 mg/day/4 days). Urinary steroid profiles in the elderly males were compared to the steroid patterns found in 15 healthy young men (age 28.9+/-5.1 years). In the females the results were compared to their individual baseline excretion without dex pretreatment. Urinary steroid determinations were carried out by semiautomatic capillary gas-liquid chromatography. In both genders DHEA administration induced significant increases in urinary DHEA (females: baseline vs. 50 mg vs. 100 mg: 361+/-131 vs. 510+/-264 vs. 1541+/-587 microg/day; males: placebo vs. 50 mg vs. 100 mg: 434+/-154 vs. 1174+/-309 vs. 4751+/-1059 microg/day) as well as in the major DHEA metabolites androsterone (A) and etiocholanolone (Et). Fifty mg DHEA led to an excretion of DHEA and its metabolites only slightly above baseline levels found in young females and in young men, respectively, whereas 100 mg induced clearly supraphysiological values. After 50 mg DHEA the ratios of urinary DHEA metabolites (A/DHEA, Et/DHEA) were not significantly different between elderly males vs. young male volunteers and young healthy females versus their individual baseline levels. In conclusion, an oral dose of 30 to 50 mg DHEA restores a physiological urinary steroid profile in subjects with DHEA deficiency without evidence for a relevant hepatic first-pass effect on urinary metabolites.
Assuntos
Desidroepiandrosterona/administração & dosagem , Desidroepiandrosterona/metabolismo , Administração Oral , Adulto , Androgênios/metabolismo , Androgênios/farmacocinética , Androgênios/urina , Dexametasona/farmacologia , Relação Dose-Resposta a Droga , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores Sexuais , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Tetra-Hidrocortisona/urinaRESUMO
The interconversion of hormonally active cortisol (F) and inactive cortisone (E) is catalyzed by two isozymes of 11beta-hydroxysteroid dehydrogenase (11betaHSD), an oxo-reductase converting E to F (11betaHSD1) and a dehydrogenase (11betaHSD2) converting F to E. 11betaHSD1 is important in mediating glucocorticoid-regulated glucose homeostasis and regional adipocyte differentiation. Earlier studies conducted with GH-deficient subjects treated with replacement GH suggested that GH may modulate 11betaHSD1 activity. In 7 acromegalic subjects withdrawing from medical therapy (Sandostatin-LAR; 20-40 mg/month for at least 12 months), GH rose from 7.1 +/- 1.5 to 17.5 +/- 4.3 mU/L (mean +/- SE), and insulin-like growth factor I (IGF-I) rose from 43.0 +/- 8.8 to 82.1 +/- 13.7 nmol/L (both P < 0.05) 4 months after treatment. There was a significant alteration in the normal set-point of F to E interconversion toward E. The fall in the urinary tetrahydrocortisols/tetrahydocortisone ratio (THF+allo-THF/THE; 0.82 +/- 0.06 to 0.60 +/- 0.06; P < 0.02) but unaltered urinary free F/urinary free E ratio (a marker for 11betaHSD2 activity) suggested that this was due to inhibition of 11betaHSD1 activity. An inverse correlation between GH and the THF+allo-THF/THE ratio was observed (r = -0.422; P < 0.05). Conversely, in 12 acromegalic patients treated by transsphenoidal surgery (GH falling from 124 +/- 49.2 to 29.3 +/- 15.4 mU/L; P < 0.01), the THF+allo-THF/THE ratio rose from 0.53 +/- 0.06 to 0.63 +/- 0.07 (P < 0.05). Patients from either group who failed to demonstrate a change in GH levels showed no change in the THF+allo-THF/THE ratio. In vitro studies conducted on cells stably transfected with either the human 11betaHSD1 or 11betaHSD2 complementary DNA and primary cultures of human omental adipose stromal cells expressing only the 11betaHSD1 isozyme indicated a dose-dependent inhibition of 11betaHSD1 oxo-reductase activity with IGF-I, but not GH. Neither IGF-I nor GH had any effect on 11betaHSD2 activity. GH, through an IGF-I-mediated effect, inhibits 11betaHSD1 activity. This reduction in E to F conversion will increase the MCR of F, and care should be taken to monitor the adequacy of function of the hypothalamo-pituitary-adrenal axis in acromegalic subjects and in GH-deficient, hypopituitary patients commencing replacement GH therapy. Conversely, enhanced E to F conversion occurs with a reduction in GH levels; in liver and adipose tissue this would result in increased hepatic glucose output and visceral adiposity, suggesting that part of the phenotype currently attributable to adult GH deficiency may be an indirect consequence of its effect on tissue F metabolism via 11betaHSD1 expression.
Assuntos
Hormônio do Crescimento Humano/farmacologia , Hidroxiesteroide Desidrogenases/metabolismo , Fator de Crescimento Insulin-Like I/farmacologia , Isoenzimas/metabolismo , 11-beta-Hidroxiesteroide Desidrogenases , Acromegalia/tratamento farmacológico , Acromegalia/metabolismo , Acromegalia/cirurgia , Tecido Adiposo/enzimologia , Adulto , Idoso , Diferenciação Celular , Linhagem Celular , Feminino , Hormônio do Crescimento Humano/metabolismo , Humanos , Fator de Crescimento Insulin-Like I/metabolismo , Rim/enzimologia , Masculino , Pessoa de Meia-Idade , Octreotida/administração & dosagem , Adeno-Hipófise/cirurgia , Células Estromais/enzimologia , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismoRESUMO
A method is described for the preparation of multi-labeled tetrahydrocortisol (3alpha,11beta,17alpha,21-tetrahydroxy-5beta-[1, 2,3,4,5-2H5]pregnan-20-one, THF-d5), allo-tetrahydrocortisol (3alpha,11beta,17alpha,21-tetrahydroxy-5alpha-[1 ,2,3,4,5-2H5]pregnan-20-one, allo-THF-d5), and tetrahydrocortisone (3alpha,17alpha,21-trihydroxy-5beta-[1,2,3,4,5-2H5]pre gnane-11,20-dione, THE-d5) containing five non-exchangeable deuterium atoms in the steroid ring A. Reductive deuteration at C-1, C-2, C-3, C-4, and C-5 of prednisolone or prednisone was performed in CH3COOD with rhodium (5%) on alumina under the deuterium atmosphere. The isotopic purities of the labeled compounds as [2H5]-form were estimated to be 86.17 atom%D for THF-d5, 74.46 atom%D for allo-THF-d5 and 81.90 atom%D for THE-d5, based on the ion intensities in the region of the molecular ion of methoxime-trimethylsilyl (MO-TMS) derivatives measured by GC-MS.
Assuntos
Hidrocortisona/metabolismo , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismo , Cromatografia Líquida , Deutério , Humanos , Marcação por Isótopo , Espectroscopia de Ressonância Magnética , Espectrometria de MassasRESUMO
A reversed-phase high-performance liquid chromatography-electrospray-tandem mass spectrometry assay (HPLC-ESI-MS/MS) was developed to quantitate cortisol, cortisone, 20 alpha- and beta-dihydrocortisol, 20 alpha- and beta-dihydrocortisone, tetrahydrocortisol, and tetrahydrocortisone. The technique was used to analyze perfusate from the isolated human placental lobule for cortisol and its metabolites. Analytes were prepared from the perfusion medium using C18 solid-phase extraction cartridges. The internal standard for the analyses was 6 alpha-methylprednisolone. Chromatography was performed on a Novapak C18 column at ambient temperature using 53% methanol and 47% 10 mM ammonium formate buffer (pH 4.0) as mobile phase, at a flow rate of 80 microL/min. A PE-SCIEX API III triple quadrupole instrument was used for mass spectrometric detection. An ionspray (pneumatically assisted electrospray) interface was used in negative and positive ionization mode. The assay was linear over the range 100-2000 micrograms/L for each analyte. The instrumental limit of detection was 50 pg. Assay imprecision at 400 and 800 micrograms/L was < or = 10% (total coefficient of variation). Accuracy ranged between 83.2% for 20 beta-dihydrocortisone to 102.6% for cortisone. Recovery of 1000 micrograms/L analyte ranged from 91.3% for cortisone to 109.7% for tetrahydrocortisol.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Hidrocortisona/análise , Espectrometria de Massas/métodos , Placenta/química , 11-beta-Hidroxiesteroide Desidrogenases , Cortisona/análogos & derivados , Cortisona/análise , Cortisona/metabolismo , Estudos de Avaliação como Assunto , Feminino , Humanos , Hidrocortisona/análogos & derivados , Hidrocortisona/metabolismo , Hidroxiesteroide Desidrogenases/antagonistas & inibidores , Técnicas In Vitro , Perfusão , Placenta/metabolismo , Gravidez , Tetra-Hidrocortisol/análise , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/análise , Tetra-Hidrocortisona/metabolismoRESUMO
A characteristic feature of the ectopic ACTH syndrome is a state of mineralocorticoid excess, although the etiology remains obscure. Some forms of endocrine hypertension, such as licorice ingestion, have been explained by cortisol acting as a mineralocorticoid in the setting of inhibition or deficiency of 11 beta-hydroxysteroid dehydrogenase (11 beta HSD). This enzyme is responsible for the conversion of cortisol (F) to hormonally inactive cortisone, and its activity in vivo can be inferred from the ratio of the urinary excretion of tetrahydrocortisol (THF) and its isomer (5 alpha THF) to tetrahydrocortisone. Twenty-two patients with Cushing's syndrome (11 pituitary dependent, 9 ectopic, and 2 adrenal adenomas) and 13 controls were studied. Compared to controls. Cushing's patients had a significant increase (P < 0.001) in the excretion of all principal metabolites of F, secondary to a 5- to 6-fold increase in the cortisol secretion rate [median, 34.0 (range, 13.3-327) mg/day in Cushing's vs. 6.1 (range, 2.5-10.3) mg/day in controls]. The THF plus 5 alpha THF/tetrahydrocortisone ratio was significantly increased in Cushing's syndrome regardless of etiology [mean, 1.81 (range, 1.09-9.99) in Cushing's vs. 0.81 (range, 0.51-1.47) in controls; P < 0.001), indicative of defective 11 beta HSD activity. Furthermore, compared to patients with pituitary-dependent Cushing's, this ratio was significantly higher in patients with the ectopic ACTH syndrome (4.12 vs. 1.49; P < 0.01) and was inversely correlated with serum potassium levels (r = -0.57; P = 0.01; n = 22). One explanation for the mineralocorticoid excess state of the ectopic ACTH syndrome appears to be that cortisol gains inappropriate access to the mineralocorticoid receptor through failure of its normal metabolism by 11 beta HSD. The reason for the defective 11 beta HSD activity is unclear, but it may be secondary to substrate saturation, inhibition by other adrenal steroids, or product inhibition.
Assuntos
Síndrome de ACTH Ectópico/metabolismo , Síndrome de Cushing/metabolismo , Hidroxiesteroide Desidrogenases/metabolismo , Mineralocorticoides/metabolismo , 11-beta-Hidroxiesteroide Desidrogenases , Feminino , Humanos , Hidrocortisona/metabolismo , Masculino , Pessoa de Meia-Idade , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismoRESUMO
During continuous infusion of 3H-cortisol in the circulation of the guinea-pig mother or fetus, radioactive metabolites appear in both maternal and fetal blood. These cortisol-derived compounds were identified principally as cortisone, tetrahydrocortisol (THF) and tetrahydrocortisone (THE). There were unidentified others in low quantities. The cortisone of the maternal plasma is 100% maternal in origin since that of the fetal plasma is 50% fetal in origin between days 62 and 66 and increased thereafter. An identical profile was noted for THF. THE seemed to be synthetized in the fetal guinea-pig and was transferred to the mother in increasing amounts near term. Liver concentrations of cortisol were higher than those of plasma in the mother. Maternal liver appeared to be the main organ of cortisol metabolism in the mother-fetus unit, but maternal adrenal may contribute to this metabolism.
Assuntos
Hidrocortisona/metabolismo , Troca Materno-Fetal , Glândulas Suprarrenais/metabolismo , Animais , Cortisona/metabolismo , Feminino , Feto/metabolismo , Cobaias , Hidrocortisona/sangue , Fígado/metabolismo , Placenta/metabolismo , Gravidez , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisona/metabolismoRESUMO
In order to determine the possibility of assessing intrauterine fetal well being by measuring amniotic fluid steroid levels, a number of selected glucocorticoids and estrogens were assayed. Estriol (E3) and estetrol (E4) levels at 36-40 weeks of pregnancy, especially that of total E3 (T-E3 and unconjugated E4 (U-E4) rose significantly (p less than 0.01), and were at a ratio of T-E3/U-E3 8; T-E4/U-E4 2, respectively. A definite correlation was found between T-E3 and U-E3 (r = 0.97) which was not detected in the case of E4, and there was a high correlation with umbilical arterial levels of both unconjugated and total E3 and E4. Amniotic fluid cortisol (F) levels in both unconjugated (U-F) and sulfate conjugated (S-F) rose preferentially at 36-40 weeks and exceeded the levels of both umbilical arterial and maternal peripheral plasma (p less than 0.01). Both tetrahydro-cortisone (THE) and -cortisol (THF) levels increased at 29-35 weeks which in the select cases were comparable with the increases seen at 36-40 weeks. The THE level showed a definite correlation with that of umbilical artery (r = 0.73) whereas this was not the case for S-F. In pregnancies complicated with toxemia and/or diabetes mellitus, T-E3 and S-F levels are less than the lower limits for normal pregnancies. This tendency was not observed with T-E4 and THE. In pregnancies with severe RH isoimmunization, the levels of T-E3, T-E4 and S-F are markedly lower than those seen in normal pregnancies, and do not display the increase with gestational advances. On the other hand, THE and THF levels were shown to be at levels higher than in normal pregnancies. Thus, in the latter half of pregnancy, amniotic fluid steroid assays, especially T-E3, S-F and THE may be of use in the assessment of fetal well being.
Assuntos
Líquido Amniótico/metabolismo , Cortisona/análogos & derivados , Estetrol/metabolismo , Estriol/análogos & derivados , Estriol/metabolismo , Tetra-Hidrocortisona/metabolismo , Feminino , Sangue Fetal/metabolismo , Humanos , Pré-Eclâmpsia/metabolismo , Gravidez , Complicações na Gravidez/metabolismo , Terceiro Trimestre da Gravidez , Gravidez em Diabéticas/metabolismoRESUMO
6 alpha-Hydroxy metabolites of cortisol were determined in the urine of pregnant (36-40 weeks of gestation) and nonpregnant women and in amniotic fluid from nearly fullterm pregnant women because relatively large amounts of these compounds are excreted in the urine of 2-day-old infants (> 200 micrograms/day). The corticosteroids analyzed by high pressure liquid chromatography and gas chromatography-mass spectrometry were 6 alpha-hydroxy derivatives of (allo)tetrahydrocortisone (3 alpha, 17 alpha, 21-trihydroxy-5 epsilon-pregnan-11,20-dione), (allo)tetrahydrocortisol (3 alpha, 11 beta, 17 alpha, 21-tetrahydroxy-5 epsilon-pregnan-20-one), and alpha- and beta-cortolone (3 alpha, 17 alpha, 20 epsilon, 21-tetrahydroxy-5 beta-pregnan-11-one). All of these compounds were found in the urine samples from both groups of women and in the amniotic fluid samples in contrast to those found in the urine samples from the neonates where 6 alpha-hydroxy compounds of (allo)tetrahydrocortisol and allotetrahydrocortisone were not positively identified because of insufficient yields. The pregnant women excreted significantly larger amounts of 6 alpha-hydroxy metabolites of cortisol (approximately 600 micrograms/day) than the control women (approximately 90 micrograms/day), and the rate of urinary excretion of these 6 alpha-hydroxy compounds was 7.82 and 1.30 micrograms/kg . day, respectively, for these groups of women compared to 54.3 micrograms/kg . day for the neonates. The precursors of these metabolites within the fetal body originated largely from the maternal circulation, and, therefore, the 6 alpha-hydroxy metabolites of cortisol excreted by the mother refer mainly to fetal metabolism and to a lesser extent, to the fetal secretion of cortisol.
Assuntos
Cortisona/análogos & derivados , Hidrocortisona/análogos & derivados , Recém-Nascido , Gravidez , Pregnanos/urina , Tetra-Hidrocortisol/análogos & derivados , Tetra-Hidrocortisona/análogos & derivados , Adulto , Líquido Amniótico/metabolismo , Feminino , Feto/metabolismo , Humanos , Hidroxicorticosteroides/metabolismo , Hidroxicorticosteroides/urina , Masculino , Pregnanos/metabolismo , Tetra-Hidrocortisol/metabolismo , Tetra-Hidrocortisol/urina , Tetra-Hidrocortisona/metabolismo , Tetra-Hidrocortisona/urinaRESUMO
The role of steroid-17-aldols, 20 beta-isocortisol (11 beta, 17,20 beta-trihydroxy-3-oxo-pregn-4-en-21-al) or 20 beta-iso THE (3 alpha,17,20 beta-trihydroxy-11-oxo-pregnan-21-al), as preferred intermediates for the biosynthesis of cortoic acids was studied in human subjects. The results demonstrated that the isosteroids were converted moe efficiently than cortisol to cortoic acids and hexahydro neutral metabolites. In all cases, the oxidation state at C-11 was largely conserved. After the administration of the 20 beta compounds both 20 alpha and 20 beta epimers of the acidic and neutral metabolites were isolated. This inversion occurred without oxidation at C-20 and provided evidence for the mediation of an epimerase in this transformation. The results further indicate that reversion of the isosteroids to ketolic intermediates (i.e. cortisol, tetrahydrocortisol, and tetrahydrocortisone did not occur.