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1.
J Endocrinol Invest ; 45(2): 347-359, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34341969

RESUMO

BACKGROUND: Congenital adrenal hyperplasia (CAH) due to autosomal recessive 21-hydroxylase deficiency (21-OHD) is caused by defects in the CYP21 (CYP21A2) gene. Several mutations have been identified in the CYP21 (CYP21A2) gene of patients with 21-OHD. We aimed at determining the frequency of these mutations among a group of Egyptian patients and studying the genotype-phenotype correlation. METHODS: Forty-seven patients with CAH due to 21-OHD from 42 different families diagnosed by clinical and hormonal evaluation and classified accordingly into salt wasting (SW) and simple virilizing (SV) phenotypes were enrolled. Their ages ranged between 1.78 and 18.99 years. Molecular analysis of the CYP21 (CYP21A2) gene was performed for the detection of eleven common mutations: P30L, I2 splice (I2 G), Del 8 bp E3 (G110del8nt), I172N, cluster E6 (I236N, V237E, M239K), V281L, L307 frameshift (F306 + T), Q318X, R356W, P453S, R483P by polymerase chain reaction (PCR) and reverse hybridization. RESULTS: Disease-causing mutations were identified in 47 patients, 55.31% of them were compound heterozygous. The most frequent mutations were I2 splice (25.43%), followed by cluster E6 (16.66%) and P30L (15.78%). Two point mutations (P453S, R483P) were not identified in any patient. In the SW patients, genotypes were more compatible with their phenotypes. CONCLUSION: Molecular characterization should be considered along with clinical and biochemical diagnosis of CAH since it could confirm the diagnosis, outline the treatment strategy and morbidity, and ensure proper genetic counseling.


Assuntos
Hiperplasia Suprarrenal Congênita , Cortisona/biossíntese , Esteroide 21-Hidroxilase/genética , Virilismo , Desequilíbrio Hidroeletrolítico , Hiperplasia Suprarrenal Congênita/sangue , Hiperplasia Suprarrenal Congênita/epidemiologia , Hiperplasia Suprarrenal Congênita/genética , Hiperplasia Suprarrenal Congênita/fisiopatologia , Criança , Egito/epidemiologia , Feminino , Estudos de Associação Genética/métodos , Estudos de Associação Genética/estatística & dados numéricos , Predisposição Genética para Doença , Testes Genéticos/métodos , Humanos , Lactente , Masculino , Mutação , Seleção de Pacientes , Virilismo/diagnóstico , Virilismo/epidemiologia , Virilismo/genética , Desequilíbrio Hidroeletrolítico/diagnóstico , Desequilíbrio Hidroeletrolítico/epidemiologia , Desequilíbrio Hidroeletrolítico/genética , Adulto Jovem
2.
Metab Eng ; 55: 59-67, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31212031

RESUMO

Steroidal compounds are one of the most widely marketed pharmaceutical products. Chemical synthesis of steroidal compounds faces many challenges, including the requirement for multiple chemical steps, low yield and selectivity in several synthesis steps, low profitability and the production of environmental pollutants. Consequently, in recent decades there has been growing interest in the use of microbial systems to produce pharmaceutical compounds. Several microbial systems have recently been developed for the microbial synthesis of the glucocorticoid hydrocortisone, which serves as a key intermediate in the production of several other pharmaceutically important steroidal compounds. In this study, we sought to establish an efficient, microbial-based system, for the conversion of hydrocortisone into cortisone. To this end, we developed a strategy for high-yield cortisone production based on ectopic expression of the guinea-pig 11ß-Hydroxysteroid dehydrogenase type 1 (11ß-HSD1) in Bacillus megaterium. We screened different constructs, containing a variety of promoters tailored for B. megaterium, and created modified versions of the enzyme by protein engineering to optimize cortisone yield. Furthermore, we utilized co-expression of an alcohol dehydrogenase to promote NADP+ regeneration, which significantly improved 11ß-HSD1 activity. The process thereby developed was found to show a remarkably high regioselectivity of >95% and to generate cortisone yields of up to 13.65 g L-1 d-1, which represents a ∼1000-fold improvement over the next-best reported system. In summary, we demonstrate the utility of B. megaterium MS941 as a suitable host for recombinant protein production and its high potential for industrial steroid production.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 1 , Bacillus megaterium , Cortisona/biossíntese , Hidrocortisona/metabolismo , Microrganismos Geneticamente Modificados , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/genética , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Animais , Bacillus megaterium/enzimologia , Bacillus megaterium/genética , Cortisona/genética , Cobaias , Hidrocortisona/genética , Microrganismos Geneticamente Modificados/genética , Microrganismos Geneticamente Modificados/metabolismo , Oxirredução , Engenharia de Proteínas
3.
Appl Biochem Biotechnol ; 185(4): 1004-1013, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29399745

RESUMO

ß-cyclodextrins derivatives (CDs) have applied in steroids biotransformation industry because of their unique properties. However, the effect of ß-CDs on the growth rate, activity, conversion, and characters of whole cells has not been concerned. In this study, the growth rate and cellular morphology of Arthrobacter simplex (ASP) pretreated by six kinds of ß-CDs were measured. The results showed that most ß-CDs inhibited the growth of ASP, among which randomly methylated-ß-CDs has the most serve inhibition; however, sulfonic acid-ß-CD promoted the growth of cells. The morphology size and the surface of all ß-CDs-pretreated cells were changed compared with the control group. Besides, the conversion of cortisone acetate (CA) increased in ß-CDs-pretreatment system and ß-CDs-containing system, which reached 97.98% in hydroxypropyl-ß-cyclodextrin (HP-ß-CD) containing-system and 78.69% in HP-ß-CD-pretreatment-system, but the dehydrogenase activity of all ß-CDs-pretreated cells decreased. ß-CDs with higher K value have stronger inclusion ability with CA, and along with the membrane permeability of ß-CDs-pretreated cells increased more, but they also have more serve damage on the ASP cells, which is negative to increase the conversion of CA. This study improved our understanding of the effect of ß-CDs when they were used in the steroids biotransformation by ASP whole cells, and provided data basis for the selection of suitable CDs for application.


Assuntos
Arthrobacter/metabolismo , Cortisona/análogos & derivados , beta-Ciclodextrinas/metabolismo , Biotransformação , Cortisona/biossíntese , Metilação
4.
Biotechnol Lett ; 38(4): 597-602, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26704767

RESUMO

OBJECTIVE: To establish a method for the recovery and reutilization of hydroxypropyl-ß-cyclodextrin (HP-ß-CD) to lower the cost of its industrial application in cortisone acetate bioconversion. RESULTS: HP-ß-CD is not degraded by Arthrobacter simplex CPCC140451 (ASP) resting cells and 96.4 % HP-ß-CD could be recovered by isobutyl acetate extraction. Moreover, the inclusion ability of recovered HP-ß-CD barely decreased. The saccharide metabolic and catalytic activities of ASP were greater in the aqueous phase after extracting with isobutyl acetate than other organic solvents. Cyclic utilization tests showed that cortisone acetate conversion ratio was 91.0 % after eight cycles and reached 95.7 % with 0.2-0.6 mM HP-ß-CD. Furthermore, >90 % conversion ratio was reached per cycle through a co-cyclic-utilization method with HP-ß-CD and immobilized ASP. CONCLUSION: Cortisone acetate conversion ratio in the HP-ß-CD cyclic-utilization method is promising for industrial applications. The method can also be expanded to other CDs and other hydrophobic compounds bioconversion.


Assuntos
Arthrobacter/enzimologia , Cortisona/análogos & derivados , beta-Ciclodextrinas/química , 2-Hidroxipropil-beta-Ciclodextrina , Arthrobacter/metabolismo , Proteínas de Bactérias/metabolismo , Biocatálise , Cortisona/biossíntese
5.
Toxicol Sci ; 121(2): 320-7, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21427057

RESUMO

There is increasing concern over the risk of environmentally relevant doses of bisphenol A (BPA) on human endocrine systems. Effects of BPA on steroidogenesis and the related molecular mechanisms were investigated in H295R human adenocarcinoma cells. This immortal cell line is unique in expressing all the enzymes of the steroidogenic pathways. The effects of BPA on steroidogenesis, 17ß-estradiol (E2) metabolism, and aromatase activity were examined in H295R cells exposed to BPA from 3.0 × 10(-1) to 3.0 × 10(3) ng/ml. Concentrations of BPA in basic cell culture materials were verified. Stable CYP17A-knockdown H295R cells were developed to verify the mechanism of inhibited steroidogenesis by BPA. Background concentrations of BPA in control cell culture media ranged from 0.03 to 0.38 ng/ml. Significantly lesser concentrations of androstenedione, testosterone, cortisol, and cortisone were caused by exposure to 30-3000 ng BPA/ml. In contrast, sconcentrations of estrone (E1) and E2 were significantly greater in BPA-exposed H295R cells. Lesser production of androstenedione and testosterone by H295R cells exposed to BPA was the most sensitive endpoint (no observable effect concentrations < 30 ng BPA/ml). CYP17A knockdown in H295R cells resulted in less production of both 17α hydroxyprogesterone and androstenedione. The results are consistent with the hypothesis that in H295R cells, BPA selectively inhibits 17,20-lyase but not 17α-hydroxylase. The primary mechanism causing increased E2 in the medium was inhibition of E2 metabolism rather than greater aromatase (CYP19) activity. These results suggest that BPA has the potential to interfere with cellular steroidogenesis in humans through multiple molecular mechanisms.


Assuntos
Inibidores da Aromatase/toxicidade , Aromatase/metabolismo , Disruptores Endócrinos/toxicidade , Estradiol/biossíntese , Fenóis/toxicidade , 17-alfa-Hidroxiprogesterona/metabolismo , Androstenodiona/biossíntese , Aromatase/genética , Compostos Benzidrílicos , Linhagem Celular Tumoral , Cortisona/biossíntese , Estradiol/genética , Estrona/biossíntese , Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Hidrocortisona/biossíntese , Reação em Cadeia da Polimerase , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Esteroide 17-alfa-Hidroxilase/genética , Esteroide 17-alfa-Hidroxilase/metabolismo , Testosterona/biossíntese
6.
Prikl Biokhim Mikrobiol ; 47(1): 50-7, 2011.
Artigo em Russo | MEDLINE | ID: mdl-21442921

RESUMO

Transformation of 16 delta5-3beta-hydroxy- and delta4-3-ketosteroids of androstane and pregnane classes was carried out using Curvularia lunata mycelium suspended in phosphate buffer with methyl-beta-cyclodextrine (MCD). As the result, 20 monohydroxy- and dihydroxy-metabolites, whose structure was determined using specters of proton magnetic resonance and mass-specters, have been isolated. Hydroxylation of delta5-3beta-hydroxy-steroids occurred mostly in the C-7alpha position whereas hydroxylation of delta4-3-ketosteroids was in the C-11beta position. Only androst-4-en-3,17-dione, 9alpha-hydroxyl-androstenedione, and androsts-1,4-diene-3,17-dione were hydroxylated at C-14alpha position. Besides main 11beta-derivatives, the 6beta- and 7beta-hydroxy-derivatives with yield 10 and 30%, respectively, were isolated during transformation of progesterone and hydroxymethyl pregnadienon. The ratio of MCD to transforming steroid was 1 : 1 (mol/mol). Hydroxycortisone and 7alpha-hydroxyandrostenolone with the yield 55 and 77%, respectively, were obtained at the maximal concentrations of cortexolone 20 g/l and androstenolone acetate 10 g/l in the presence of MCD. Absorption of steroids on mycelium, lower speed of their transformation, low concentrations of modifying substrates, and low yield of hydroxyderivatives have been observed in the absence of MCD.


Assuntos
Cortisona/biossíntese , Desidroepiandrosterona/biossíntese , Hidroxiesteroides/metabolismo , Cetosteroides/metabolismo , beta-Ciclodextrinas/metabolismo , Ascomicetos/química , Ascomicetos/metabolismo , Técnicas de Cultura de Células , Cortisona/análogos & derivados , Cortisona/isolamento & purificação , Desidroepiandrosterona/análogos & derivados , Desidroepiandrosterona/isolamento & purificação , Hidroxilação , Hidroxiesteroides/química , Cetosteroides/química , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Micélio/química , Micélio/metabolismo , Fosfatos/metabolismo , Solubilidade
7.
Rev Med Chil ; 136(6): 701-10, 2008 Jun.
Artigo em Espanhol | MEDLINE | ID: mdl-18769825

RESUMO

BACKGROUND: Cortisol has been implicated in hypertension and lately reported to be regulated at the pre-receptor level by the 11betaHSD1 enzyme, which converts cortisone (E) to cortisol (F). Over-expression of this enzyme in adipose tissue could determine an increase in available cortisol that interacts with the mineralocorticoid receptor (MR) in renal, brain and heart tissue, leading to similar hypertensive effects as in 11betaHSD2 impaired patients. Several polymorphisms have been reported in HSDl IB 1 gene (CAI5, CAI9 and InsA83557), which could modify HSDl IB 1 gene expression or activity. AIM: To determine the distribution and prevalence of CAI5, CAI9 and InsA83557 in the HSDl IBl gene, and to correlate these results with biochemical parameters in cortisol/ ACTH (HPA) and renin-angiotensin-aldosterone (RAA) axis in patients with essential hypertension (EH). PATIENTS AND METHODS: We studied 113 EH patients (76 non-obese and 37 obese, with a body mass índex >30 kg/m(2)) and 30 normotensive adults (NT). In each patient, we measured serum levels of E E, serum aldosterone (SA), plasma renin activity (PRA), adrenocorticotrophic hormone (ACTH), the urinary free cortisol/creatinine (UFF/Cr), F/ACTH and SA/PRA ratios. Each polymorphism was studied by PCR and 8% polyacrylamide gel electrophoresis. Statistical associations were evaluated by Pearson correlations and the genetic equilibrium by the Hardy-Weinberg (H-W) equation. RESULTS: We found all three polymorphisms in the EH and the NT group, both in genetic equilibrium. In obese essential hypertensives, the CAI5 polymorphism showed association with SA/PRA ratio (r =0.189, p =0.012) and F/ACTH (r =0.301, p 0.048); CA19 also showed correlation with F/ACTH in obese EH (r = 0.220, p 0.009). The InsA83557polymorphism correlated with UFF/Cr in both EH (r =0.206; p =0.03), and in obese EH (r =0.354; p =0.05). CONCLUSIONS: The CAI5 and CAI9 polymorphism correlated with changes in biochemical parameters in HPA and RAA axis of obese essential hypertensives. These changes may result in modifications in the expression of 11betaHSD1, leading to increased cortisol and aldosterone levels independent of ACTH and renin control, respectively.


Assuntos
Hipertensão/genética , Polimorfismo Genético , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/genética , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/metabolismo , Hormônio Adrenocorticotrópico/sangue , Adulto , Aldosterona/sangue , Estudos de Casos e Controles , Doença Crônica , Cortisona/biossíntese , Feminino , Frequência do Gene , Humanos , Hidrocortisona/sangue , Hipertensão/enzimologia , Masculino , Repetições de Microssatélites , Obesidade/enzimologia , Obesidade/genética , Reação em Cadeia da Polimerase , Renina/sangue , Adulto Jovem
8.
Rev. méd. Chile ; 136(6): 701-710, jun. 2008. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-490754

RESUMO

Background: Cortisol has been implicated in hypertension and lately reported to be regulated at the pre-receptor level by the 11ßHSD1 enzyme, which converts cortisone (E) to cortisol (F). Over expression ofthis enzyme in adipose tissue could determine an increase in available cortisol that interacts with the mineralocorticoid receptor (MR) in renal, brain and heart tissue, leading to similar hypertensive effects as in 11ßHSD2 impaired patients. Severa! polymorphisms have been reported in HSDl IB 1 gene (CAI5, CAI9 and InsA83557), which could modify HSDl IB 1 gene expression or activity. Aun: To determine the distribution and prevalence of CAI5, CAI9 and InsA83557 in the HSDl IBl gene, and to correlate these results with biochemical parameters in cortisol/ ACTH (HPA) and renin-angiotensin-aldosterone (RAA) axis in patients with essential hypertension (EH). Patients and Methods: We studied 113 EHpatients (76 non-obese and 37 obese, with a body mass índex >30 kg/m²) and 30 normotensive adults (NT). In each patient, we measured serum levéis of E E, serum aldosterone (SA), plasma renin activity (PRA), adrenocorticotrophic hormone (ACTH), the urinary free cortisol/creatinine (UFF/Cr), F/ACTH and SA/PRA ratios. Each polymorphism was studied by PCR and 8 percent polyacrylamide gel electrophoresis. Statistical associations were evaluated by Pearson correlations and the genetic equilibñum by the Hardy-Weinberg (H-W) equation. Results: We found all three polymorphisms in the EH and the NT group, both in genetic equilibñum. In obese essential hypertensives, the CAI5polymorphism showed association with SA/PRA ratio (r =0.189, p =0.012) and F/ACTH (r =0.301, p 0.048); CA19 also showed correlation with F/ACTH in obese EH (r = 0.220, p 0.009). The InsA83557polymorphism correlated with UFF/Cr in both EH (r =0.206; p =0.03), and in obese EH (r =0.354; p =0.05). Conclusions: The CAI5 and CAI9 polymorphism correlated with changes in biochemical parameters...


Assuntos
Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Hipertensão/genética , Polimorfismo Genético , /genética , /metabolismo , Hormônio Adrenocorticotrópico/sangue , Aldosterona/sangue , Estudos de Casos e Controles , Doença Crônica , Cortisona/biossíntese , Frequência do Gene , Hidrocortisona/sangue , Hipertensão/enzimologia , Repetições de Microssatélites , Obesidade/enzimologia , Obesidade/genética , Reação em Cadeia da Polimerase , Renina/sangue , Adulto Jovem
9.
J Endocrinol ; 195(1): 1-6, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17911391

RESUMO

The year 1946 was not only the year that the Society for Endocrinology was founded, but also the year that Edward Kendall's compound E (cortisone) was first synthesised by Louis Sarett. By 1948, sufficient quantities of compound E were available for the rheumatologist Philip Hench to test it successfully for the first time in a patient with rheumatoid arthritis. It was immediately hailed as a 'wonder drug' and was shown to be effective in a number of inflammation-associated conditions, most notably rheumatoid arthritis. The subsequent development of endocrinology as a discipline is inextricably linked to the chemistry, biology and medicine of antiinflammatory glucocorticoids. Sixty years after the first chemical synthesis of cortisone, corticosteroids remain among the top ten most commonly used prescription and over the counter drugs. Basic and clinical studies of glucocorticoid biosynthesis, metabolism and action have trail-blazed developments in endocrinology ever since. This article surveys the extraordinary cortisone timeline, from first synthesis until now. The concluding scientific message is that intracrine metabolism of cortisone to cortisol via 11beta hydroxysteroid dehydrogenase type 1 likely sustains local amplification of glucocorticoid action at sites of inflammation throughout the body. The broader message is that the discovery of compound E by Kendall (basic scientist), its large-scale synthesis by Sarett (industrial chemist) and its therapeutic application by Hench (rheumatologist) serves as a paradigm for modern translational medicine. It is concluded that endocrinology will remain a force in health and disease if it continues to evolve sans frontières at the basic/applied/clinical science interface. A challenge for the Society for Endocrinology is to ensure this happens.


Assuntos
Cortisona/biossíntese , Cortisona/uso terapêutico , Endocrinologia/tendências , Anti-Inflamatórios/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/metabolismo , Humanos , Hidrocortisona/metabolismo
10.
J Pineal Res ; 41(1): 58-66, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16842542

RESUMO

The primate fetal adrenal reaches a large size relative to body weight followed by a rapid decrease in size in the postnatal period. We tested the hypothesis that maternal melatonin stimulates growth and prevents maturation of the primate fetal adrenal gland. We suppressed maternal melatonin by exposing eight pregnant capuchin monkeys to constant light (LL) from 63% to 90% gestation (term 155 days). Three of these received daily oral melatonin replacement (LL + Mel). Five mothers remaining in light:dark cycle were used as controls. Fetuses were delivered at 90% gestation. The absence of maternal melatonin selectively decreased fetal adrenal weight (Control: 488.8 +/- 51.5; LL: 363.2 +/- 27.7 and LL + Mel 519 +/- 46 mg; P < 0.05 ANOVA) without effecting fetal weight, placental weight or the weight of other fetal tissues. Changes in fetal adrenal size were accompanied by an increase in the levels of Delta5-3beta-hydroxysteroid dehydrogenase (3beta-HSD) mRNA (Control: 0.8 +/- 0.2; LL: 5.2 +/- 0.6 and LL + Mel 0.8 +/- 0.1; 3beta-HSD/18S-rRNA; P < 0.05 ANOVA). In vitro we found that maternal melatonin suppression increased basal progesterone production to levels similar to those of the adult adrenal gland (Control: 0.36 +/- 0.09; LL 0.99 +/- 0.13; LL + Mel 0.18 +/- 0.06 and adult: 0.88 +/- 0.10 ng/mg of tissue; P < 0.05 ANOVA) but no change in cortisol production. We found an increased production of cortisone (Control: 1.65 +/- 0.60; LL: 5.44 +/- 0.63; LL + Mel: 2.90 +/- 0.38 and adult: 1.70 +/- 0.45 ng/mg of tissue; P < 0.05 ANOVA). Collectively, the effects of maternal melatonin suppression and their reversion by maternal melatonin replacement suggest that maternal melatonin stimulates growth and prevents maturation of the capuchin monkey fetal adrenal gland.


Assuntos
Glândulas Suprarrenais/embriologia , Desenvolvimento Fetal/fisiologia , Melatonina/fisiologia , 3-Hidroxiesteroide Desidrogenases/genética , 3-Hidroxiesteroide Desidrogenases/metabolismo , Glândulas Suprarrenais/enzimologia , Glândulas Suprarrenais/metabolismo , Animais , Cebus , Cortisona/biossíntese , DNA Complementar , Feminino , Hidrocortisona/biossíntese , Imuno-Histoquímica , Masculino , Progesterona/biossíntese , RNA Mensageiro/genética
11.
Diabetes ; 55(3): 667-74, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16505229

RESUMO

The splanchnic bed produces cortisol at rates approximating extraadrenal tissues by converting cortisone to cortisol via the 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 pathway. It is not known whether splanchnic cortisol production is regulated by nutrient ingestion and/or by the accompanying changes in hormone secretion. To address this question, 18 healthy humans were randomized to ingest either a mixed meal or to receive an intravenous saline infusion while total-body, splanchnic, and D3 cortisol production (an index of 11beta-HSD type 1 activity) were measured using the combined hepatic catheterization and D4 cortisol infusion methods. Fasting glucose and insulin concentrations did not differ on the meal and saline study days. Glucose and insulin concentrations increased after meal ingestion, peaking at 11.0 +/- 1.0 mmol/l and 451 +/- 64 pmol/l, respectively, at 45 min, then fell to baseline thereafter. In contrast, glucose and insulin concentrations slowly fell to 5.1 +/- 0.1 mmol/l and 27 +/- 6 pmol/l during the 6 h of observation on the saline study day. Fasting cortisol concentration did not differ on the meal and saline study days. Cortisol increased (P < 0.05) to a peak of 353 +/- 55 nmol/l after meal ingestion but did not change after saline infusion. The increase in cortisol after meal ingestion was associated with an increase in both total body cortisol (from 748 +/- 63 to 1,620 +/- 235 nmol/min; P < 0.01) and total body D3 cortisol (from 99 +/- 11 to 143 +/- 11 nmol/min; P < 0.01) production, whereas there was no change in either on the saline study day. The increase in total-body cortisol and D3 cortisol production after meal ingestion originated in extrasplanchnic tissues since splanchnic cortisol production (mean 0-360 min: 254 +/- 83 vs. 262 +/- 36 nmol/min) and splanchnic D3 cortisol production (mean 0-360 min: 72 +/- 22 vs. 77 +/- 14 nmol/min) did not differ on the meal and saline study days. We conclude that ingestion of a mixed meal does not alter either splanchnic cortisol production or the conversion of D4 cortisol to D3 cortisol or, therefore by implication, flux via the splanchnic 11beta-HSD type 1 pathway.


Assuntos
Hidrocortisona/biossíntese , Circulação Esplâncnica , 11-beta-Hidroxiesteroide Desidrogenases/fisiologia , Adulto , Glicemia/análise , Cortisona/biossíntese , Feminino , Artéria Femoral/metabolismo , Alimentos , Veias Hepáticas/metabolismo , Humanos , Insulina/sangue , Masculino , Pessoa de Meia-Idade
12.
J Clin Endocrinol Metab ; 88(6): 2384-92, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12788832

RESUMO

11 beta-Hydroxysteroid dehydrogenase type 2 (11 beta-HSD2) plays a crucial role in converting hormonally active cortisol to inactive cortisone, thereby conferring specificity on the mineralocorticoid receptor. Mutations in the gene encoding 11 beta-HSD2 (HSD11B2) account for an inherited form of hypertension, the syndrome of apparent mineralocorticoid excess, in which cortisol induces hypertension and hypokalemia. A similar clinical picture to apparent mineralocorticoid excess occurs after the ingestion of licorice and carbenoxolone, which are competitive inhibitors of 11 beta-HSD2. Reduced 11 beta-HSD2 activity may explain the increased sodium retention in preeclampsia, renal disease, and liver cirrhosis. Substrate saturation of 11 beta-HSD2 occurs in Cushing's syndrome and explains the mineralocorticoid excess state that characterizes ectopic ACTH syndrome. Polymorphic variability in the HSD11B2 gene in part determines salt sensitivity, a forerunner for adult onset hypertension. Furthermore, reduced placental 11 beta-HSD2 expression might underpin the Barker hypothesis, the epidemiological link between reduced birth weight and adult hypertension. At a prereceptor level, 11 beta-HSD2 plays a key role in normal physiology in the corticosteroid regulation of sodium homeostasis and pathophysiology of hypertension.


Assuntos
Cortisona/biossíntese , Hidrocortisona/metabolismo , Hidroxiesteroide Desidrogenases/metabolismo , Hipertensão/metabolismo , 11-beta-Hidroxiesteroide Desidrogenase Tipo 2 , Humanos , Hipertensão/etiologia , Mineralocorticoides/metabolismo
13.
J Clin Endocrinol Metab ; 81(11): 3855-60, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8923828

RESUMO

We describe two female siblings who had production of cortisol (F; as determined from excretion of urinary metabolites) high enough to give rise to Cushing's disease, but who had no clinical indications of the condition. The teenage patients were hirsute as a result of adrenal hyperandrogenism. A notable feature of the condition was the elevated excretion of corticosteroid metabolites with 11-carbonyl groups and very low excretion of 11 beta-hydroxylated steroids. We termed this disorder apparent cortisone (E) reductase disorder. The steroid metabolite phenotype appeared to be the opposite of that seen in the apparent mineralocorticoid excess syndrome, in which the excretion of 11-keto compounds is attenuated. As an example, the tetrahydrocortisol plus 5 alpha-tetrahydrocortisol/tetrahydrocortisone ratio was about 0.04 compared to normal values of about 1.0 and apparent mineralocorticoid excess syndrome values of 5.0-50.0. Paradoxically, among the F metabolites that had not undergone A-ring reduction, 11 beta-hydroxylated steroids dominated over 11-carbonyl compounds. The F/E ratio was about 1.8 compared to an average normal value of 0.54. Neither the father nor the mother of the patient had abnormal F metabolite/E metabolite ratios, although the father did excrete highly elevated free E and F, possibly an unrelated condition. A conclusion was not reached regarding the basis of the disorder. We considered that the most likely causes were 1) defective hepatic 11 beta-hydroxysteroid dehydrogenase-1, 2) failure to develop the adult form of F metabolism, or 3) excessive activity of A ring reduction enzymes acting on E.


Assuntos
Hiperfunção Adrenocortical/enzimologia , Hidrocortisona/biossíntese , Hidroxiesteroide Desidrogenases/deficiência , 11-beta-Hidroxiesteroide Desidrogenases , Adolescente , Corticosteroides/metabolismo , Hiperfunção Adrenocortical/etiologia , Hiperfunção Adrenocortical/metabolismo , Adulto , Androgênios/biossíntese , Cortisona/biossíntese , Cortisona/metabolismo , Feminino , Hirsutismo/enzimologia , Hirsutismo/etiologia , Hirsutismo/metabolismo , Humanos , Hidrocortisona/metabolismo , Hidroxiesteroide Desidrogenases/metabolismo , Fígado/enzimologia , Masculino , Fenótipo
14.
J Steroid Biochem Mol Biol ; 49(1): 81-5, 1994 May.
Artigo em Inglês | MEDLINE | ID: mdl-8003443

RESUMO

Inhibition of 11 beta-hydroxysteroid dehydrogenase (11 beta-OHSD) can cause excess mineralocorticoid effects and hypokalemia. Several substances causing hypokalemia (glycyrrhizic acid in licorice and gossypol) inhibit this enzyme. We tested other compounds for activity to inhibit 11 beta-OHSD in guinea pig kidney cortex microsomes with NADP as cofactor and cortisol as substrate. Furosemide was an inhibitor while bumetanide was not, indicating a mechanism for the increase K+ excretion caused by furosemide compared with bumetanide. Naringenin (found in grapefruit juice), ethacrynic acid, and chenodeoxycholic acid had inhibitor IC50 values similar to glycyrrhizic acid. We conclude that various compounds can inhibit this enzyme and may play a role in K+ metabolism and adrenocorticosteroid action.


Assuntos
Flavanonas , Flavonoides/farmacologia , Furosemida/farmacologia , Hidroxiesteroide Desidrogenases/antagonistas & inibidores , Córtex Renal/enzimologia , Microssomos/enzimologia , 11-beta-Hidroxiesteroide Desidrogenases , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Ácidos Cólicos/farmacologia , Cortisona/biossíntese , Antagonistas de Estrogênios/farmacologia , Ácido Glicirretínico/análogos & derivados , Ácido Glicirretínico/farmacologia , Ácido Glicirrízico , Cobaias , Hidrocortisona/metabolismo , Técnicas In Vitro , Córtex Renal/efeitos dos fármacos , Cinética , Masculino , Microssomos/efeitos dos fármacos , Esteróis/farmacologia
15.
J Pharm Pharmacol ; 46(4): 305-9, 1994 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8051615

RESUMO

To identify the inhibitor of prednisolone metabolism contained in Saiboku-To, we conducted in-vitro experiments of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), using rat liver homogenate and cortisol as a typical substrate. We studied the effects of ten herbal constituents on 11 beta-HSD. Five herbal extracts showed inhibitory activity with Glycyrrhiza glabra > Perillae frutescens > Zizyphus vulgaris > Magnolia officinalis > Scutellaria baicalensis. This suggests that unknown 11 beta-HSD inhibitors are contained in four herbs other than G. glabra which contains a known inhibitor, glycyrrhizin (and glycyrrhetinic acid). Seven chemical constituents which have been identified as the major urinary products of Saiboku-To in healthy and asthmatic subjects were studied; magnolol derived from M. officinalis showed the most potent inhibition of the enzyme (IC50, 1.8 x 10(-4) M). Although this activity was less than that of glycyrrhizin, the inhibition mechanism (non-competitive) was different from a known competitive mechanism. These results suggest that magnolol might contribute to the inhibitory effects of Saiboku-To on prednisolone metabolism through inhibition of 11 beta-HSD.


Assuntos
Asma/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Hidroxiesteroide Desidrogenases/antagonistas & inibidores , Imunossupressores/farmacologia , Lignanas , Fígado/efeitos dos fármacos , Medicina Kampo , 11-beta-Hidroxiesteroide Desidrogenases , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Antineoplásicos Fitogênicos/uso terapêutico , Compostos de Bifenilo/isolamento & purificação , Compostos de Bifenilo/farmacologia , Compostos de Bifenilo/uso terapêutico , Cromatografia Líquida de Alta Pressão , Cortisona/biossíntese , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/uso terapêutico , Glucocorticoides/análise , Ácido Glicirretínico/análogos & derivados , Ácido Glicirretínico/farmacologia , Ácido Glicirrízico , Hidrocortisona/metabolismo , Imunossupressores/química , Imunossupressores/uso terapêutico , Fígado/enzimologia , Masculino , Ratos , Ratos Wistar
16.
J Endocrinol Invest ; 16(1): 65-8, 1993 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8445158

RESUMO

The in vitro metabolism of [1,2-3H] deoxycorticosterone (DOC), [1,2-3H] 18-hydroxy-11-deoxycorticosterone (18-OHDOC) and [1,2-3H] 11-deoxycortisol (S) was studied in adrenal adenoma homogenates from patients with primary hyperaldosteronism. Tumor tissues actively converted deoxycorticosterone and 18-hydroxy-11-deoxycorticosterone to 18-hydroxycorticosterone and aldosterone. Yields of cortisol and cortisone were also large showing that the tissues did not lack the zona fasciculata-like 11 beta-hydroxylation ability.


Assuntos
Adenoma/metabolismo , Neoplasias das Glândulas Suprarrenais/metabolismo , Glucocorticoides/biossíntese , Hiperaldosteronismo/metabolismo , Mineralocorticoides/biossíntese , 18-Hidroxicorticosterona/metabolismo , 18-Hidroxidesoxicorticosterona/metabolismo , Aldosterona/biossíntese , Cortisona/biossíntese , Cortodoxona/metabolismo , Desoxicorticosterona/metabolismo , Feminino , Humanos , Hidrocortisona/biossíntese , Masculino
17.
Chronobiol Int ; 9(5): 350-5, 1992 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-1394606

RESUMO

The circadian rhythm of the in vitro biosynthesis of cortisol and cortisone in mice adrenals has been documented in the absence and presence of 0.1 mumol metyrapone, an inhibitor of steroid 11 beta-monooxygenase. After 3 weeks of synchronization with 12 h light:12 h darkness, adrenalectomy was performed at eight circadian stages: 0, 4, 9, 10, 13, 16, 21, and 22 h after light onset (HALO). Because it has been shown that mice adrenals could convert exogenous 11-deoxycortisol, the synthesis of 11-oxysteroids (cortisol+cortisone) in adrenal homogenates was studied from tritiated precursor. The pattern of steroid synthesis showed a maximum around the end (10 HALO) and a minimum at the beginning of the resting period (0 HALO); the variation was approximately 10%. A similar pattern was observed in the presence of a approximately 50% inhibiting dose of metyrapone. On the other hand, the percent inhibition of 11-oxysteroids synthesis was greater at the beginning of the resting period (0 HALO) and minimum around the end of the activity span (21 HALO), with an overall variation of 20%. However, the variations were statistically insignificant (unpaired t test).


Assuntos
Glândulas Suprarrenais/efeitos dos fármacos , Ritmo Circadiano/fisiologia , Metirapona/farmacologia , Glândulas Suprarrenais/metabolismo , Animais , Cortisona/biossíntese , Hidrocortisona/biossíntese , Técnicas In Vitro , Luz , Masculino , Camundongos , Esteroide 11-beta-Hidroxilase/antagonistas & inibidores
18.
Artigo em Inglês | MEDLINE | ID: mdl-1360370

RESUMO

1. The conversion of progesterone to 17 alpha-hydroxyprogesterone (17 alpha-OHP), 11-deoxycortisol (11-DOCR) and deoxycorticosterone (DOC) was significantly higher in female rainbow trout than in male trout; in contrast, the interrenal production of cortisol (CR) plus cortisone (CN) was higher in males than in females. 2. Following treatment with 1 mg/kg of HCB, the interrenal conversion of progesterone to 17 alpha-OHP and 11-DOCR was significantly increased in male and female trout but at 20 mg/kg of HCB, the production of these metabolites was increased in males and decreased in females; CR+CN production was unchanged after HCB treatment in both sexes.


Assuntos
Glândula Inter-Renal/efeitos dos fármacos , Bifenilos Policlorados/toxicidade , Esteroides/biossíntese , 17-alfa-Hidroxiprogesterona , Animais , Autorradiografia , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Cortisona/biossíntese , Cortodoxona/metabolismo , Desoxicorticosterona/biossíntese , Feminino , Hidrocortisona/biossíntese , Hidroxiprogesteronas/metabolismo , Glândula Inter-Renal/metabolismo , Masculino , Progesterona/metabolismo , Caracteres Sexuais , Truta/metabolismo
19.
Acta Endocrinol (Copenh) ; 125(2): 160-4, 1991 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-1897333

RESUMO

To study the possible impairment of 11 beta-hydroxysteroid-dehydrogenase in patients with chronic renal insufficiency, urinary excretion rates of the four main glucocorticoid-metabolites, tetrahydrocortisol, tetrahydrocortisone, allotetrahydrocortisol and allotetrahydrocortisone were determined by capillary gas chromatography in 22 patients with chronic renal insufficiency with (N = 15) and without (N = 7) hypertension, but without hemodialysis treatment. Whereas the sum of all 41 steroid metabolites determined by capillary gas chromatography was reduced (p less than 0.001) in patients with chronic renal insufficiency as compared with 22 healthy individuals, the relative contribution of the four glucocorticoid metabolites to total steroid excretion was similar in patients with renal insufficiency (22 +/- 12%) and in healthy subjects (20 +/- 5%). However, the excreted amount of tetrahydrocortisol exceeded that of tetrahydrocortisone in all but 3 (normotensive) patients with chronic renal insufficiency, but only in one healthy subject resulting, in patients with chronic renal insufficiency, in a ratio of tetrahydrocortisone vs tetrahydrocortisone of 0.7 +/- 0.4 (hypertensive patients 0.5 +/- 0.2; normotensive patients 1.1 +/- 0.4; controls 1.9 +/- 0.9, p less than 0.001 vs patients with chronic renal insufficiency). This ratio of tetrahydrocortisone/tetrahydrocortisol showed a correlation with serum concentrations of creatinine (p less than 0.001). These results provide indirect proof of an impaired conversion of cortisol to cortisone in moderate renal insufficiency and may suggest a relationship with the hypertension frequently seen in this group of patients.


Assuntos
Cortisona/biossíntese , Hidrocortisona/metabolismo , Hipertensão/etiologia , Falência Renal Crônica/metabolismo , Adulto , Idoso , Cromatografia Gasosa , Feminino , Glucocorticoides/urina , Humanos , Falência Renal Crônica/complicações , Falência Renal Crônica/urina , Masculino , Pessoa de Meia-Idade , Testosterona/metabolismo
20.
Biochem Int ; 20(6): 1111-8, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-2164396

RESUMO

Snake (Ptyas mucosa) pituitaries were heated, extracted with an acidic medium, and the extract subsequently chromatographed on carboxymethyl cellulose (CMC). Fractions were assayed for their abilities to displace D-ala2-D-leu5-[tyrosyl, 3,53H] enkephalin from binding to rat brain membranes and 125I-beta-endorphin from binding to its antibody, and to stimulate corticosterone production by isolated rat adrenal cells. The fraction unadsorbed on CMC (having the least basic character) had the lowest opiate receptor binding activity and beta-endorphin-like immunoreactivity. The highest steroidogenic activity, opiate receptor binding activity and beta-endorphin-like immunoreactivity were concentrated in a strongly adsorbed fraction. Snake pituitaries were also extracted with Tris-HCl buffer (pH 7.8). The extract from the second procedure was subsequently chromatographed on ConA-Sepharose, ultrafiltered and dialyzed to obtain a nonglycopeptide fraction with a molecular weight between 1,000 and 10,000. This fraction also exhibited steroidogenic and opiate receptor binding activities.


Assuntos
Hormônio Adrenocorticotrópico/análise , Hipófise/metabolismo , Serpentes/metabolismo , beta-Endorfina/análise , Hormônio Adrenocorticotrópico/metabolismo , Animais , Cromatografia por Troca Iônica , Cortisona/biossíntese , Masculino , Radioimunoensaio , Ratos , Ratos Endogâmicos , Receptores Opioides/metabolismo , beta-Endorfina/metabolismo
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