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1.
J Med Invest ; 68(3.4): 238-243, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34759137

RESUMO

Peripherally derived steroids affect steroid production in the brain via the blood-brain barrier. However, steroid concentrations are lower in the cerebrospinal fluid than those in the blood, indicating restricted influx of steroids because of their metabolization by choroid plexus (CP) epithelial cells. Here, we analyzed the gene expression of steroidogenic enzymes [cholesterol side-chain cleavage enzyme (P450scc), 17α-hydroxylase/C17-C20 lyase (P450c17), 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase type 1 (17ß-HSD1), aromatase (Cyp19a1), and 5α-reductase type 1 (5α-R1)]. These genes were expressed to a lesser extent in the CP than in the testis and to a similar extent in the cerebral cortex. However, P450scc levels were higher in the CP than in the cerebral cortex, whereas Cyp19a1 levels showed the opposite trend. We also evaluated the effects of orchiectomy and testosterone on the expression of these genes. P450c17 and 5α-R1 levels were unaffected by orchiectomy, whereas P450scc and 3ß-HSD levels were increased and decreased, respectively. Cyp19a1 expression increased upon testosterone treatment, whereas that of 17ß-HSD decreased upon orchiectomy or administration of testosterone. Immunohistochemistry analysis revealed that 17ß-HSD was expressed in the cytoplasm of CP epithelial cells. These results indicate that CP epithelial cells synthesize and convert the certain types of steroids to contribute to the homeostasis of steroids in the brain. J. Med. Invest. 68 : 238-243, August, 2021.


Assuntos
Enzima de Clivagem da Cadeia Lateral do Colesterol , Plexo Corióideo , Animais , Enzima de Clivagem da Cadeia Lateral do Colesterol/genética , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Plexo Corióideo/metabolismo , Masculino , Ratos , Esteroide 17-alfa-Hidroxilase/genética , Esteroide 17-alfa-Hidroxilase/metabolismo , Esteroides , Testosterona
2.
J Steroid Biochem Mol Biol ; 212: 105924, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34089832

RESUMO

Endogenous Cushing syndrome (CS) is an endocrine disorder marked by excess cortisol production rendering patients susceptible to visceral obesity, dyslipidemia, hypertension, osteoporosis and diabetes mellitus. Adrenal CS is characterized by autonomous production of cortisol from cortisol-producing adenomas (CPA) via adrenocorticotropic hormone-independent mechanisms. A limited number of studies have quantified the steroid profiles in sera from patients with CS. To understand the intratumoral steroid biosynthesis, we quantified 19 steroids by mass spectrometry in optimal cutting temperature compound (OCT)-embedded 24 CPA tissue from patients with overt CS (OCS, n = 10) and mild autonomous cortisol excess (MACE, n = 14). Where available, normal CPA-adjacent adrenal tissue (AdjN) was also collected and used for comparison (n = 8). Immunohistochemistry (IHC) for CYP17A1 and HSD3B2, two steroidogenic enzymes required for cortisol synthesis, was performed on OCT sections to confirm the presence of tumor tissue and guided subsequent steroid extraction from the tumor. LC-MS/MS was used to quantify steroids extracted from CPA and AdjN. Our data indicated that CPA demonstrated increased concentrations of cortisol, cortisone, 11-deoxycortisol, corticosterone, progesterone, 17OH-progesterone and 16OH-progesterone as compared to AdjN (p < 0.05). Compared to OCS, MACE patient CPA tissue displayed higher concentrations of corticosterone, 18OH-corticosterone, 21-deoxycortisol, progesterone, and 17OH-progesterone (p < 0.05). These findings also demonstrate that OCT-embedded tissue can be used to define intra-tissue steroid profiles, which will have application for steroid-producing and steroid-responsive tumors.


Assuntos
Adenoma/metabolismo , Neoplasias do Córtex Suprarrenal/metabolismo , Síndrome de Cushing/metabolismo , Esteroides/metabolismo , Adenoma/sangue , Neoplasias do Córtex Suprarrenal/sangue , Adulto , Cromatografia Líquida , Síndrome de Cushing/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Progesterona Redutase/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Esteroides/sangue , Espectrometria de Massas em Tandem
3.
Ecotoxicol Environ Saf ; 217: 112198, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33862428

RESUMO

The mechanism of neurodevelopmental toxicity of decabromodiphenyl ether (BDE209) remains unclear. Recent evidence suggests that neurosteroids disorders play a vital role in BDE209 induced-neurodevelopmental toxicity. To explore the mechanism of it, pregnant ICR mice were orally gavaged with 0, 225, and 900 mg kg-1 BDE209 for about 42 days. Spatial learning and memory abilities of offspring were tested on postnatal day (PND) 21. Offspring were euthanized at PND26, the neuronal structure, neurosteroids level, and related proteins including neurosteroids synthase, ionotropic receptors and cAMP-response element binding protein (CREB) pathway were evaluated, as well as Ca2+ concentration and the mitochondrial membrane potential (Mmp). Our results showed that BDE209 impaired learning and memory abilities and disrupted neuronal structure. Meanwhile, BDE209 decreased the pregnenolone (PREG), dehydroepiandrosterone (DHEA), progesterone (PROG) and allopregnanolone (ALLO) levels in the serum and brain, as well as the mRNA and protein levels of cholesterol-side-chain cleavage enzyme (P450scc), steroid 17α-hy-droxylase (P450C17), 3ß-hydroxysteroid dehydrogenase (3ß-HSD) and steroid 5α-reductase of type I (5α-R) in the hippocampi. Also, BDE209 suppressed mRNA and protein levels of NR1, NR2A and NR2B subunits of the N-methyl-D-aspartic acid receptor (NMDAR) and α1 subunit of the Gamma-amino butyric acid A receptor (GABAAR), but increased the levels of ß2 and γ2 subunits of the GABAAR in the hippocampi. Moreover, BDE209 increased the Ca2+ concentration and phosphorylation extracellular regulated protein kinases (P-ERK) 1/2 level, but decreased the P-CREB and Mmp level in the hippocampi. These results indicate that BDE209 exposure during pregnancy and lactation is possible to affect learning and memory formation of offspring by the neurosteroid-mediated ionotropic receptors dysfunction.


Assuntos
Éteres Difenil Halogenados/toxicidade , Sistema Nervoso/crescimento & desenvolvimento , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/crescimento & desenvolvimento , Encéfalo/metabolismo , Enzima de Clivagem da Cadeia Lateral do Colesterol , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Feminino , Lactação , Masculino , Camundongos , Camundongos Endogâmicos ICR , Sistema Nervoso/efeitos dos fármacos , Neurônios/metabolismo , Neuroesteroides , Gravidez , Pregnanolona/metabolismo , Progesterona/metabolismo , Receptores de GABA/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo
4.
Nat Commun ; 12(1): 2260, 2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33859207

RESUMO

Metabolic control is mediated by the dynamic assemblies and function of multiple redox enzymes. A key element in these assemblies, the P450 oxidoreductase (POR), donates electrons and selectively activates numerous (>50 in humans and >300 in plants) cytochromes P450 (CYPs) controlling metabolism of drugs, steroids and xenobiotics in humans and natural product biosynthesis in plants. The mechanisms underlying POR-mediated CYP metabolism remain poorly understood and to date no ligand binding has been described to regulate the specificity of POR. Here, using a combination of computational modeling and functional assays, we identify ligands that dock on POR and bias its specificity towards CYP redox partners, across mammal and plant kingdom. Single molecule FRET studies reveal ligand binding to alter POR conformational sampling, which results in biased activation of metabolic cascades in whole cell assays. We propose the model of biased metabolism, a mechanism akin to biased signaling of GPCRs, where ligand binding on POR stabilizes different conformational states that are linked to distinct metabolic outcomes. Biased metabolism may allow designing pathway-specific therapeutics or personalized food suppressing undesired, disease-related, metabolic pathways.


Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Ligantes , Redes e Vias Metabólicas , Aromatase/metabolismo , Linhagem Celular , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/isolamento & purificação , Ensaios Enzimáticos , Transferência Ressonante de Energia de Fluorescência , Humanos , Lipossomos/metabolismo , Simulação de Acoplamento Molecular , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Imagem Individual de Molécula , Esteroide 17-alfa-Hidroxilase/metabolismo , Esteroide 21-Hidroxilase/metabolismo , Especificidade por Substrato
5.
J Am Chem Soc ; 143(10): 3729-3733, 2021 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-33656879

RESUMO

CYP17A1 is an essential human steroidogenic enzyme, which catalyzes two sequential reactions leading to the formation of androstenedione from progesterone and dehydroepiandrosterone from pregnenolone. The second reaction is the C17-C20 bond scission, which is strongly dependent on the presence of cytochrome b5 and displays a heretofore unexplained more pronounced acceleration when 17OH-progesteone (17OH-PROG) is a substrate. The origin of the stimulating effect of cytochrome b5 on C-C bond scission catalyzed by CYP17A1 is still debated as mostly due to either the acceleration of the electron transfer to the P450 oxy complex or allosteric effects of cytochrome b5 favoring active site conformations that promote lyase activity. Using resonance Raman spectroscopy, we compared the effect of Mn-substituted cytochrome b5 (Mn-Cytb5) on the oxy complex of CYP17A1 with both proteins co-incorporated in lipid nanodiscs. For CYP17A1 with 17OH-PROG, a characteristic shift of the Fe-O mode is observed in the presence of Mn-b5, indicating reorientation of a hydrogen bond between the 17OH group of the substrate from the terminal to the proximal oxygen atom of the Fe-O-O moiety, a configuration favorable for the lyase catalysis. For 17OH-pregnenolone, no such shift is observed, the favorable H-bonding orientation being present even without Mn-Cytb5. These new data provide a precise allosteric interpretation for the more pronounced acceleration seen for the 17OH-PROG substrate.


Assuntos
Citocromos b5/química , Esteroide 17-alfa-Hidroxilase/metabolismo , Regulação Alostérica , Biocatálise , Domínio Catalítico , Citocromos b5/metabolismo , Humanos , Pregnenolona/química , Pregnenolona/metabolismo , Esteroide 17-alfa-Hidroxilase/química , Especificidade por Substrato
6.
J Biol Chem ; 296: 100571, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33753170

RESUMO

It has been recognized for >50 years that cytochrome b5 (b5) stimulates some cytochrome P450 (P450)-catalyzed oxidations, but the basis of this function is still not understood well. The strongest stimulation of catalytic activity by b5 is in the P450 17A1 lyase reaction, an essential step in androgen synthesis from 21-carbon (C21) steroids, making this an excellent model system to interrogate b5 function. One of the issues in studying b5-P450 interactions has been the limited solution assay methods. We constructed a fluorescently labeled variant of human b5 that can be used in titrations. The labeled b5 bound to WT P450 17A1 with a Kd of 2.5 nM and rapid kinetics, on the order of 1 s-1. Only weak binding was observed with the clinical P450 17A1 variants E305G, R347H, and R358Q; these mutants are deficient in lyase activity, which has been hypothesized to be due to attenuated b5 binding. Kd values were not affected by the presence of P450 17A1 substrates. A peptide containing the P450 17A1 Arg-347/Arg-358 region attenuated Alexa 488-T70C-b5 fluorescence at higher concentrations. The addition of NADPH-P450 reductase (POR) to an Alexa 488-T70C-b5:P450 17A1 complex resulted in a concentration-dependent partial restoration of b5 fluorescence, indicative of a ternary P450:b5:POR complex, which was also supported by gel filtration experiments. Overall, these results are interpreted in the context of a dynamic and tight P450 17A1:b5 complex that also binds POR to form a catalytically competent ternary complex, and variants that disrupt this interaction have low catalytic activity.


Assuntos
Androgênios/biossíntese , Citocromos b5/metabolismo , Liases/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Humanos , Cinética , Mutação , Ligação Proteica , Esteroide 17-alfa-Hidroxilase/genética
7.
Mol Cell Endocrinol ; 528: 111261, 2021 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-33781841

RESUMO

Cytochrome P450 17A1 (CYP17A1) is a critical steroidogenic enzyme, essential for producing glucocorticoids and sex hormones. This review discusses the complex activity of CYP17A1, looking at its role in both the classical and backdoor steroidogenic pathways and the complex chemistry it carries out to perform both a hydroxylation reaction and a carbon-carbon cleavage, or lyase reaction. Functional and structural investigations have informed our knowledge of these two reactions. This review focuses on a few specific aspects of this discussion: the identities of reaction intermediates, the coordination of hydroxylation and lyase reactions, the effects of cytochrome b5, and conformational selection. These discussions improve understanding of CYP17A1 in a physiological setting, where CYP17A1 is implicated in a variety of steroidogenic diseases. This information can be used to improve ways in which CYP17A1 can be effectively modulated to treat diseases such as prostate and breast cancer, Cushing's syndrome, and glioblastoma.


Assuntos
Neoplasias da Mama/metabolismo , Síndrome de Cushing/metabolismo , Glioblastoma/metabolismo , Neoplasias da Próstata/metabolismo , Esteroide 17-alfa-Hidroxilase/química , Esteroide 17-alfa-Hidroxilase/metabolismo , Domínio Catalítico , Feminino , Humanos , Hidroxilação , Masculino , Especificidade por Substrato
8.
J Clin Endocrinol Metab ; 106(5): 1389-1397, 2021 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-33524149

RESUMO

BACKGROUND: While previous studies indicate that the zonae reticularis (ZR) and glomerulosa (ZG) diminish with aging, little is known about age-related transformations of the zona fasciculata (ZF). OBJECTIVES: To investigate the morphological and functional changes of the adrenal cortex across adulthood, with emphasis on (i) the understudied ZF and (ii) sexual dimorphisms. METHODS: We used immunohistochemistry to evaluate the expression of aldosterone synthase (CYP11B2), visinin-like protein 1 (VSNL1), 3ß-hydroxysteroid dehydrogenase type II (HSD3B2), 11ß-hydroxylase (CYP11B1), and cytochrome b5 type A (CYB5A) in adrenal glands from 60 adults (30 men), aged 18 to 86. Additionally, we employed mass spectrometry to quantify the morning serum concentrations of cortisol, 11-deoxycortisol (11dF), 17α-hydroxyprogesterone, 11-deoxycorticosterone, corticosterone, and androstenedione in 149 pairs of age- and body mass index-matched men and women, age 21 to 95 years. RESULTS: The total cortical area was positively correlated with age (r = 0.34, P = 0.008). Both the total (VSNL1-positive) and functional ZG (CYP11B2-positive) areas declined with aging in men (r = -0.57 and -0.67, P < 0.01), but not in women. The CYB5A-positive area declined with age in both sexes (r = -0.76, P < 0.0001). In contrast, the estimated ZF area correlated positively with age in men (r = 0.59, P = 0.0006) and women (r = 0.49, P = 0.007), while CYP11B1-positive area remained unchanged across ages. Serum cortisol, corticosterone, and 11-deoxycorticosterone levels were stable across ages, while 11dF levels increased slightly with age (r = 0.16, P = 0.007). CONCLUSION: Unlike the ZG and ZR, the ZF and the total adrenal cortex areas enlarge with aging. An abrupt decline of the ZG occurs with age in men only, possibly contributing to sexual dimorphism in cardiovascular risk.


Assuntos
Córtex Suprarrenal/patologia , Córtex Suprarrenal/fisiologia , Envelhecimento/fisiologia , Adolescente , Córtex Suprarrenal/metabolismo , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/patologia , Estudos de Casos e Controles , Citocromo P-450 CYP11B2/metabolismo , Citocromos b5/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Tamanho do Órgão , Progesterona Redutase/metabolismo , Esteroide 11-beta-Hidroxilase/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Adulto Jovem , Zona Fasciculada/metabolismo , Zona Fasciculada/patologia , Zona Glomerulosa/metabolismo , Zona Glomerulosa/patologia , Zona Reticular/metabolismo , Zona Reticular/patologia
9.
Mol Cell Endocrinol ; 526: 111194, 2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33592286

RESUMO

This study demonstrates the application of a mathematical steroidogenic model, constructed with individual in vitro enzyme characterisations, to simulate in vivo steroidogenesis in a diseased state. This modelling approach was applied to the South African Angora goat, that suffers from hypocortisolism caused by altered adrenal function. These animals are extremely vulnerable to cold stress, leading to substantial monetary loss in the mohair industry. The Angora goat has increased CYP17A1 17,20-lyase enzyme activity in comparison with hardy livestock species. Determining the effect of this altered adrenal function on adrenal steroidogenesis during a cold stress response is difficult. We developed a model describing adrenal steroidogenesis under control conditions, and under altered steroidogenic conditions where the animal suffers from hypocortisolism. The model is parameterised with experimental data from in vitro enzyme characterisations of a hardy control species. The increased 17,20-lyase activity of the Angora goat CYP17A1 enzyme was subsequently incorporated into the model and the response to physiological stress is simulated under both control and altered adrenal steroidogenic conditions.


Assuntos
Hidrocortisona/metabolismo , Modelos Moleculares , Esteroide 17-alfa-Hidroxilase/metabolismo , Esteroides/biossíntese , Animais , Simulação por Computador , Cabras , Funções Verossimilhança , Reprodutibilidade dos Testes , Fatores de Tempo
10.
Anim Reprod Sci ; 226: 106694, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33486154

RESUMO

An electromagnetic field (EMF) may have effects on female reproduction. This study was conducted to determine whether EMF [50 and 120 Hz, 2 and 4 h of incubation in the presence or absence of progesterone (P4, 10-5 M)] affects androgen synthesis and release from the pig endometrium. Endometrial slices were collected from pigs (n = 5) during the fetal peri-implantation period (i.e., days 15-16 of gestation) and treated in vitro with EMF. The selected endometrial slices were treated with P4 to determine whether this hormone has effects on protection of the tissue from EMF radiation. The CYP17A1 and HSD3B1 mRNA transcript abundance, steroid 17αhydroxylase/17, 20-lyase (cytochrome P450c17) and hydroxyΔ5steroid dehydrogenase/3ß and steroidΔisomerase (3ßHSD) protein abundance were examined using Real-Time PCR and Western Blot procedures, respectively. In media collected after incubation, the concentrations of androstenedione (A4) and testosterone (T) were quantified used a RIA. When P4 was added to the culture medium, EMF radiation had suppressive effects on endometrial T release after 2 and 4 h of incubation when the EMF treatment was occurring and increased A4 release after 4 h of incubation with EMF at 120 Hz. When there was no inclusion of P4, release of A4 was decreased after 2 h of EMF treatment at 120 Hz and after 4 h of EMF treatment at 50 and 120 Hz. Progesterone did not have functions that protected the pig endometrium against EMF radiation during the fetal peri-implantation period.


Assuntos
Campos Eletromagnéticos/efeitos adversos , Implantação do Embrião/efeitos da radiação , Endométrio/efeitos da radiação , Suínos/fisiologia , Testosterona/metabolismo , Animais , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Gravidez , Progesterona/metabolismo , Progesterona Redutase/genética , Progesterona Redutase/metabolismo , Esteroide 17-alfa-Hidroxilase/genética , Esteroide 17-alfa-Hidroxilase/metabolismo , Esteroide Isomerases/genética , Esteroide Isomerases/metabolismo
11.
Mol Cell Endocrinol ; 525: 111174, 2021 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-33503463

RESUMO

Advances in technology are only beginning to reveal the complex interactions between hosts and their resident microbiota that have co-evolved over centuries. In this review, we present compelling evidence that implicates the host-associated microbiome in the generation of 11ß-hydroxyandrostenedione, leading to the formation of potent 11-oxy-androgens. Microbial steroid-17,20-desmolase cleaves the side-chain of glucocorticoids (GC), including cortisol (and its derivatives of cortisone, 5α-dihydrocortisol, and also (allo)- 3α, 5α-tetrahydrocortisol, but not 3α-5ß-tetrahydrocortisol) and drugs (prednisone and dexamethasone). In addition to side-chain cleavage, we discuss the gut microbiome's robust potential to transform a myriad of steroids, mirroring much of the host's metabolism. We also explore the overlooked role of intestinal steroidogenesis and efflux pumps as a potential route for GC transport into the gut. Lastly, we propose several health implications from microbial steroid-17,20-desmolase function, including aberrant mineralocorticoid, GC, and androgen receptor signaling in colonocytes, immune cells, and prostate cells, which may exacerbate disease states.


Assuntos
Bactérias/enzimologia , Trato Gastrointestinal/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Animais , Microbioma Gastrointestinal , Saúde , Humanos , Hidrocortisona/química , Hidrocortisona/metabolismo
13.
Ecotoxicol Environ Saf ; 208: 111427, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33049449

RESUMO

This study aimed to determine the in vivo effect of silver nanoparticles (AgNPs) on the concentration of sex steroids (progesterone - P4, estradiol - E2, testosterone - T) and thyroid hormones (thyroxine - T4, triiodothyronine - T3) in the blood plasma as well as the messenger ribonucleic acid (mRNA) and protein expression of HSD3ß, CYP17A1 and CYP19A1 enzymes and steroid hormone concentrations in chicken ovarian follicles. AgNPs did not affect serum steroid hormone levels, but increased T3 levels depending on the size and concentration of AgNPs. At the level of ovarian tissues, AgNPs: (i) affected the levels of E2 and T in prehierachical follicles; (ii) reduced the expression of CYP19A1 mRNA and protein and consequently diminished E2 concentration in small white follicles; and (iii) increased the expression of CYP17A1 mRNA in large white follicles, without changing its protein expression. The results indicate that AgNPs affect chicken ovarian steroidogenesis. The effects of AgNPs depend on exposure time, the type of follicle and the degree of its development and are associated with the modulation of steroidogenic gene expression and E2 and T synthesis. Prehierachical follicles seem to be more susceptible to AgNPs than preovulatory ones. In conclusion, AgNPs by targeting the chicken ovary may indirectly influence the selection processes of prehierarchical follicles to the pre-ovulatory hierarchy and disturb the ovarian steroidogenesis. Furthermore, AgNPs may affect thyroid hormone metabolism in different ways by size which in turn may influence energy homeostasis of the target cells.


Assuntos
Nanopartículas Metálicas/toxicidade , Folículo Ovariano/fisiologia , Prata/toxicidade , Hormônios Tireóideos/fisiologia , Animais , Aromatase , Galinhas/metabolismo , Estradiol/metabolismo , Feminino , Hormônios Esteroides Gonadais/metabolismo , Folículo Ovariano/efeitos dos fármacos , Ovário/metabolismo , Progesterona/metabolismo , RNA Mensageiro/metabolismo , Prata/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Testosterona/metabolismo , Hormônios Tireóideos/metabolismo , Tri-Iodotironina/metabolismo
14.
Int J Mol Sci ; 22(1)2020 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-33375507

RESUMO

The present study was performed to clone and characterize the structures and functions of steroidogenic factor 1 (sf-1) and 17α-hydroxylase/lyase (cyp17α) promoters in yellow catfish Pelteobagrus fulvidraco, a widely distributed freshwater teleost. We successfully obtained 1981 and 2034 bp sequences of sf-1 and cyp17α promoters, and predicted the putative binding sites of several transcription factors, such as Peroxisome proliferator-activated receptor alpha (PPARα), Peroxisome proliferator-activated receptor gamma (PPARγ) and Signal transducer and activator of transcription 3 (STAT3), on sf-1 and cyp17α promoter regions, respectively. Overexpression of PPARγ significantly increased the activities of sf-1 and cyp17α promoters, but overexpression of PPARα significantly decreased the promoter activities of sf-1 and cyp17α. Overexpression of STAT3 reduced the activity of the sf-1 promoter but increased the activity of the cyp17α promoter. The analysis of site-mutation and electrophoretic mobility shift assay suggested that the sf-1 promoter possessed the STAT3 binding site, but did not the PPARα or PPARγ binding sites. In contrast, only the PPARγ site, not PPARα or STAT3 sites, was functional with the cyp17α promoter. Leptin significantly increased sf-1 promoter activity, but the mutation of STAT3 and PPARγ sites decreased leptin-induced activation of sf-1 promoter. Our findings offered the novel insights into the transcriptional regulation of sf-1 and cyp17α and suggested leptin regulated sf-1 promoter activity through STAT3 site in yellow catfish.


Assuntos
Peixes-Gato/genética , Regulação da Expressão Gênica/genética , Regiões Promotoras Genéticas , Esteroide 17-alfa-Hidroxilase/genética , Fator Esteroidogênico 1/genética , Animais , Sítios de Ligação , Peixes-Gato/metabolismo , Clonagem Molecular , Genes Reporter , Células HEK293 , Humanos , Leptina/metabolismo , Luciferases/metabolismo , Mutação , PPAR alfa/genética , PPAR alfa/metabolismo , PPAR gama/genética , PPAR gama/metabolismo , Ligação Proteica , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Fator Esteroidogênico 1/metabolismo , Regulação para Cima
15.
Int J Mol Sci ; 21(19)2020 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-32992734

RESUMO

In a healthy female reproductive system, a subtle hormonal and metabolic dance leads to repetitive cyclic changes in the ovaries and uterus, which make an effective ovulation and potential implantation of an embryo possible. However, that is not so in the case of polycystic ovary syndrome (PCOS), in which case the central mechanism responsible for entraining hormonal and metabolic rhythms during the menstrual cycle is notably disrupted. In this review we provide a detailed description of the possible scenario of PCOS pathogenesis. We begin from the analysis of how a set of genetic disorders related to PCOS leads to particular malfunctions at a molecular level (e.g., increased enzyme activities of cytochrome P450 (CYP) type 17A1 (17α-hydroxylase), 3ß-HSD type II and CYP type 11A1 (side-chain cleavage enzyme) in theca cells, or changes in the expression of aquaporins in granulosa cells) and discuss further cellular- and tissue-level consequences (e.g., anovulation, elevated levels of the advanced glycation end products in ovaries), which in turn lead to the observed subsequent systemic symptoms. Since gene-editing therapy is currently out of reach, herein special emphasis is placed on discussing what kinds of drug targets and which potentially active substances seem promising for an effective medication, acting on the primary causes of PCOS on a molecular level.


Assuntos
Hormônios/metabolismo , Síndrome do Ovário Policístico , 3-Hidroxiesteroide Desidrogenases/metabolismo , Aquaporinas/metabolismo , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Feminino , Células da Granulosa/enzimologia , Células da Granulosa/patologia , Humanos , Síndrome do Ovário Policístico/tratamento farmacológico , Síndrome do Ovário Policístico/enzimologia , Síndrome do Ovário Policístico/genética , Esteroide 17-alfa-Hidroxilase/metabolismo , Células Tecais/enzimologia , Células Tecais/patologia
16.
J Clin Endocrinol Metab ; 105(12)2020 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-32865200

RESUMO

CONTEXT: Girls with premature adrenarche (PA) may have a higher risk of developing polycystic ovary syndrome (PCOS) and metabolic syndrome. The biological purpose of adrenarche is unknown and the role of novel biosynthetic pathways remains unclear. OBJECTIVE: To compare the urinary steroid metabolome and enzyme activities of girls with PA to age-matched control girls and to published steroid values of girls with normal adrenarche and of women with PCOS and their newborn daughters. DESIGN: Prospective observational study from 2009 to 2014. SETTING: Academic pediatric endocrinology referral center. PARTICIPANTS: Twenty-three girls with PA and 22 healthy, age-matched girls. MAIN OUTCOME MEASURES: Steroid metabolites in 24-hour urine samples, including 4 progesterones, 5 corticosterones, aldosterone, 13 androgens, 2 estrogens, 14 glucocorticoids, and enzyme activities represented by metabolite ratios. RESULTS: Girls with PA had a higher body mass index (mean standard deviation scores 0.9 vs -0.3, P = 0.013). Androgen excretion was higher in PA girls than in control girls (median 3257 nmol/24 hours vs 1627 nmol/24 hours, P < 0.001), in particular metabolites from alternate androgen pathways. The amount of progesterone, corticosterone, aldosterone, estrogen, and cortisol metabolites were similar between groups. Activities of 17ß-hydroxysteroid-dehydrogenase and of 17,20-lyase were higher in girls with PA. Activities of 3ß-hydroxysteroid-dehydrogenase, 21-hydroxylase, and 5α-reductase activity were not different between groups, in contrast to published results on girls with normal adrenarche or PCOS females. CONCLUSIONS: Metabolites and enzymes involved in alternate androgen pathways appear to be markers of PA. Prospective studies should assess whether steroid production in PA also differs from adrenarche at normal timing and persists into adulthood.


Assuntos
17-Hidroxiesteroide Desidrogenases/sangue , Adrenarca/sangue , Puberdade Precoce/sangue , Esteroide 17-alfa-Hidroxilase/sangue , 17-Hidroxiesteroide Desidrogenases/metabolismo , Adrenarca/metabolismo , Adrenarca/fisiologia , Androgênios/sangue , Androgênios/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Corticosterona/sangue , Corticosterona/metabolismo , Estrogênios/sangue , Estrogênios/metabolismo , Feminino , Humanos , Hidrocortisona/sangue , Hidrocortisona/metabolismo , Metaboloma , Puberdade Precoce/metabolismo , Esteroide 17-alfa-Hidroxilase/metabolismo , Suíça , Regulação para Cima
17.
J Reprod Immunol ; 142: 103191, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32937223

RESUMO

OBJECTIVE: Follicular development can be disturbed due to many factors, including having polycystic ovaries. Aberrant expression of genes involved in steroidogenesis pathway could lead to aberrant oocyte development. In this study, the gene expression levels of a number of genes that is functioning in steroidogenesis pathway were investigated. MATERIALS AND METHODS: The spare oocytes were collected from NEU Hospital IVF Center following controlled ovarian stimulation cycle. RNA was extracted using RNA/DNA Purification Kit (Norgen, Canada) and reverse transcription was performed using TruScript First Strand cDNA Synthesis Kit (Norgen, Canada). Real time PCR was conducted using LightCycler® 480 SYBR Green I Master (Roche, UK). RESULTS AND CONCLUSION: The expression levels of CYP11, CYP17, CYP19, HSD17B1, HSD3B2 and ACTB were detected in human MII stage oocytes obtained from oocyte donors aged between 18-30 years. The number of follicles and oocytes collected from the patients with polycystic ovaries were slightly higher compared to the control group. The expression level of CYP11A1 was shown to be statistically different in the oocytes obtained from the patients who do not have polycystic ovaries (p < 0.05), whereas statistically significant expression levels were observed for CYP17 in the oocytes obtained from patients with polycystic ovaries (p < 0.05). The expression level of HSD17B1 was also shown to be statistically different in the oocytes (p < 0.05). The extrapolation of the results indicates that the genes involved in steroidogenesis pathway are altered in cases of polycystic ovaries. Thus, it may have a role in the development of polycystic ovaries.


Assuntos
Androgênios/biossíntese , Hiperandrogenismo/patologia , Oócitos/enzimologia , Síndrome do Ovário Policístico/complicações , Adolescente , Adulto , Estudos de Casos e Controles , Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Estradiol Desidrogenases/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Humanos , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Oogênese , Folículo Ovariano/patologia , Síndrome do Ovário Policístico/patologia , Esteroide 17-alfa-Hidroxilase/metabolismo , Adulto Jovem
18.
Steroids ; 164: 108728, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32931809

RESUMO

Four novel indole steroids based on dehydroepiandrosterone (IS-1), estrone (IS-2) and pregnenolone (IS-3) were obtained and studied for their ability to inhibit C6 glioma proliferation. A reduction in cell proliferation by 52 ± 13% was observed for IS-1 at 10 µM, whereas IS-3 and abiraterone acetate at 10 µM caused a 36 ± 8% decrease. Surprisingly, the cellular effects reported for abiraterone, namely, cytochrome P450 CYP17A1 inhibition and endoplasmic reticulum stress were not detected for IS-1. However, both abiraterone and IS-1 significantly increased glutathione levels. Docking studies predicted good affinity of IS-1 to liver X receptors and regulatory protein Keap1, which are proposed to be involved in the compounds' antiproliferative activity.


Assuntos
Neoplasias Encefálicas/patologia , Proliferação de Células/efeitos dos fármacos , Desenho de Fármacos , Glioma/patologia , Hidroxiesteroides/farmacologia , Indóis/farmacologia , Androstenos/farmacologia , Animais , Neoplasias Encefálicas/metabolismo , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Glioma/metabolismo , Glutationa/metabolismo , Hidroxiesteroides/química , Indóis/química , Simulação de Acoplamento Molecular , Ratos , Análise Espectral/métodos , Esteroide 17-alfa-Hidroxilase/genética , Esteroide 17-alfa-Hidroxilase/metabolismo
19.
BMC Endocr Disord ; 20(1): 144, 2020 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-32957973

RESUMO

BACKGROUND: Congenital adrenal hyperplasia (CAH) with 17α-hydroxylase deficiency is a rare disease; patients often require lifetime cortisol treatment. In this case report, we presented a patient with CAH and 17α-hydroxylase deficiency, who was previously misdiagnosed as having primary aldosteronism. Furthermore, the flash glucose monitoring system (FGMS) was used to ascertain a suitable cortisol therapeutic regimen for this patient. CASE PRESENTATION: A 29-year-old woman presented with sex dysgenesis, hypertension and hypokalaemia. She had been diagnosed with primary aldosteronism at a local hospital. The re-measured aldosterone level in our hospital was below the normal range after antihypertensive medication adjustment, suggesting that the primary aldosteronism was a misdiagnosis. The patient was finally diagnosed as having CAH with 17α-hydroxylase deficiency according to the endocrine profile, adrenocorticotropic hormone stimulation test, and genetic analysis. Then, the patient was recommended cortisol treatment, during which the endocrine profile, blood pressure, plasma potassium level, and blood glucose level were observed to ascertain a suitable dosage. The FGMS was used to monitor blood glucose level, which indicated that the patient's glucose metabolism was maintained normally under the final treatment dosage. CONCLUSION: The misdiagnosis might have been because of the effects of the antihypertension medications on aldosterone and renin levels. The final dosage of cortisol treatment achieved a normal endocrine profile, while maintaining the homeostasis of blood glucose level, plasma potassium level and blood pressure. FGMS may be an effective method to ascertain a suitable cortisol therapeutic regimen for patients with CAH and 17α-hydroxylase deficiency.


Assuntos
Hiperplasia Suprarrenal Congênita/tratamento farmacológico , Automonitorização da Glicemia/métodos , Glicemia/análise , Hidrocortisona/uso terapêutico , Esteroide 17-alfa-Hidroxilase/metabolismo , Hiperplasia Suprarrenal Congênita/complicações , Hiperplasia Suprarrenal Congênita/metabolismo , Hiperplasia Suprarrenal Congênita/patologia , Hormônio Adrenocorticotrópico/metabolismo , Adulto , Anti-Inflamatórios/uso terapêutico , Feminino , Humanos , Prognóstico , Esteroide 17-alfa-Hidroxilase/genética
20.
J Steroid Biochem Mol Biol ; 204: 105750, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32920127

RESUMO

Ghrelin is a 28-amino acid peptide hormone that regulates ovarian steroid hormone synthesis; however, there is limited evidence regarding the regulation of this pathway by ghrelin in mice ovary. Thus, we aimed to investigate whether central ghrelin action plays a role in murine reproductive health by inhibiting steroid synthesis. Further, we sought to examine the mechanism of central ghrelin action in ovarian steroid hormone synthesis. After the administration of intracerebroventricular ghrelin (1 nmol), we found reduced serum concentrations of oestradiol and progesterone and reduced secretion of follicle-stimulating hormone and luteinising hormone. Although ghrelin reduced 3ß-hydroxysteroid dehydrogenase mRNA and protein levels in the hypothalamus, it did not affect the expression of steroidogenic acute regulatory protein and cytochrome P450 17A1. In the ovary, central ghrelin regulation indirectly inhibited the mRNA and protein levels of steroidogenic acute regulatory protein, cytochrome P450 17A1, and 3ß-hydroxysteroid dehydrogenase. Moreover, no changes were observed in the expression of proliferating cell nuclear antigen and phosphorylation of extracellular signal-regulated kinase. We hypothesised that central ghrelin regulation suppressed serum oestradiol and progesterone levels by indirectly inhibiting the expression of steroidogenic acute regulatory protein, cytochrome P450 17A1, and 3ß-hydroxysteroid dehydrogenase in the ovary. In this regulation, the suppressed secretion of the follicle-stimulating hormone and luteinising hormone in the pituitary by ghrelin could be involved. Furthermore, hypothalamic 3ß-hydroxysteroid dehydrogenase expression is reduced by ghrelin injection.


Assuntos
Grelina/metabolismo , Hormônios/sangue , 17-Hidroxiesteroide Desidrogenases/genética , 17-Hidroxiesteroide Desidrogenases/metabolismo , Animais , Feminino , Hipotálamo/metabolismo , Injeções Intraventriculares , Camundongos Endogâmicos C57BL , Ovário/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Reprodução , Esteroide 17-alfa-Hidroxilase/genética , Esteroide 17-alfa-Hidroxilase/metabolismo
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