Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 3.382
Filtrar
1.
J Physiol Pharmacol ; 70(3)2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31566189

RESUMO

Endocrine-disrupting chemicals (EDCs) have structures similar to steroid hormones and can interfere with hormone synthesis and normal physiological functions of reproductive organs. For example, sex steroid hormones influence calcium signaling of the cardiac muscle in early embryo development. To confirm the effect of progesterone (P4), octyl-phenol (OP), and bisphenol A (BPA) on early differentiation of mouse embryonic stem cells (mESCs) into cardiomyocytes, mESCs were treated with P4, OP, and BPA two days after attachment and media were replaced every two days. In addition, cells were treated with mifepristone (RU486), a synthetic steroid that has an affinity for progesterone receptor (Pgr), for one day starting on day 11. Beating ratio was decreased with P4, OP, and BPA treatment. The Pgr mRNA level was significantly increased in the P4-, OP- and BPA-treated groups. However, the mRNA level of the calcium channel gene (Trpv2), contraction-related genes (Ryr2, Cam2, and Mylk3) and cardiac development and morphogenesis genes (Rbp4, Ly6e, and Gata4) were significantly decreased in the P4-, OP-, and BPA-treated groups. Interestingly, treatment with RU486 rescued the altered calcium channel gene, contraction-related genes, and cardiac development and morphogenesis genes. P4, OP, and BPA treatments reduced the intracellular calcium level. Taken together, these results indicate that EDCs (OP and BPA) has a structure similar to that of endogenous steroid hormones such as progesterone and estrogen, and OP and BPA act like progesterone to inhibit and disrupt cardiomyocyte differentiation of mESCs.


Assuntos
Compostos Benzidrílicos/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Embrionárias Murinas/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Fenóis/farmacologia , Animais , Cálcio/metabolismo , Canais de Cálcio/efeitos dos fármacos , Canais de Cálcio/metabolismo , Disruptores Endócrinos/metabolismo , Estrogênios/metabolismo , Camundongos , Mifepristona/farmacologia , Células-Tronco Embrionárias Murinas/metabolismo , Miócitos Cardíacos/metabolismo , Progesterona/metabolismo , Receptores de Progesterona/metabolismo
2.
Int J Mol Sci ; 20(18)2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31510090

RESUMO

The negative association between psychological stress and male fertility has been known for many years. This study was aimed at (i) identifying spermatogenesis impairment induced by psychological stress in rats and (ii) exploring the role of glucocorticoid receptor (GR) signaling in these adverse effects (if they exist). Male Sprague Dawley rats were exposed to a six-week period of unpredictable chronic mild stress (uCMS) along with cotreatment of GR antagonist RU486 (1 mg/kg/day). Testicular damage was assessed by testicular pathological evaluation, epididymal sperm concentration, serum testosterone levels, testicular apoptotic cell measurements, and cell cycle progression analyses. Rats in the uCMS group had decreased levels of serum testosterone and decreased epididymal sperm concentration. The uCMS-treated rats also had decreased numbers of spermatids and increased levels of apoptotic seminiferous tubules; additionally, cell cycle progression of spermatogonia was arrested at the G0/G1 phase. Furthermore, uCMS exposure caused an increase in serum corticosterone level and activated GR signaling in the testes including upregulated GR expression. RU486 treatment suppressed GR signaling and alleviated the damaging effects of stress, resulting in an increased epididymal sperm concentration. Overall, this work demonstrated for the first time that the activation of GR signaling mediates stress-induced spermatogenesis impairment and that this outcome is related to cell apoptosis and cell cycle arrest in germ cells.


Assuntos
Epididimo/metabolismo , Receptores de Glucocorticoides/metabolismo , Espermatogênese/fisiologia , Espermatozoides/metabolismo , Estresse Psicológico/fisiopatologia , Testículo/metabolismo , Animais , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Epididimo/citologia , Epididimo/efeitos dos fármacos , Masculino , Mifepristona/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Glucocorticoides/antagonistas & inibidores , Túbulos Seminíferos/citologia , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/metabolismo , Contagem de Espermatozoides , Espermatogênese/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/citologia , Testículo/efeitos dos fármacos , Testosterona/sangue
3.
Biomed Khim ; 65(4): 311-315, 2019 Jun.
Artigo em Russo | MEDLINE | ID: mdl-31436172

RESUMO

It was studed basal and ACTH-stimulated production of cyclic adenosine monophosphate (cAMP) and corticosteroid hormones (progesterone and corticosterone) in rat adrenals in vitro under streptozotocin diabetes, in conditions of mifepristone administration and their combination. It was shown that in streptozotocin diabetes animals, both the basal and adrenocorticotropic hormone (ACTH) stimulated cAMP production significantly increased; this was accompanied by the increase in basal and ACTH-stimulated progesterone and corticosterone production in rat adrenals in vitro. Repeated administration of mifepristone to control and diabetic rats caused an increase mainly in ACTH-stimulated production of the main glucocorticoid hormone, corticosterone, without additional changes in the cAMP level. The results obtained suggest activation of two mechanisms of steroidogenesis enhancement in experimental animals. In rats with streptozotocin diabetes, both basal and ACTH-stimulated activity of all stages of steroidogenesis increase, which is mediated by the increased formation of cAMP as second messenger mediating the ACTH action on adrenocortical cells. Prolonged administration of mifepristone to control and diabetic rats resulted in increased activity of only late stages of steroidogenesis with predominant elevation of synthesis of physiologically active hormone corticosterone without additional changes in cAMP production level.


Assuntos
Hiperfunção Adrenocortical/fisiopatologia , AMP Cíclico/fisiologia , Diabetes Mellitus Experimental/tratamento farmacológico , Mifepristona/farmacologia , Hiperfunção Adrenocortical/complicações , Hormônio Adrenocorticotrópico/fisiologia , Animais , Corticosterona/fisiologia , Diabetes Mellitus Experimental/complicações , Ratos
4.
EBioMedicine ; 47: 170-183, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31466918

RESUMO

BACKGROUND: Recent clinical trials on ovarian cancer with mifepristone (MF) have failed, despite in vitro findings on its strong progesterone (P4) antagonist function. METHODS: Ovarian cancer human and murine cell lines, cultured high-grade human primary epithelial ovarian cancer (HG-hOEC) cells and their explants; as well as in vivo transgenic mice possessing ovarian cancer were used to assess the molecular mechanism underlying mifepristone (MF) agonistic actions in ovarian cancer progression. FINDINGS: Herein, we show that ovarian cancer cells express traceable/no nuclear P4 receptor (PGR), but abundantly P4 receptor membrane component 1 (PGRMC1). MF significantly stimulated ovarian cancer cell migration, proliferation and growth in vivo, and the translocation of PGRMC1 into the nucleus of cancer cells; the effects inhibited by PGRMC1 inhibitor. The beneficial antitumor effect of high-doses MF could not be achieved in human cancer tissue, and the low tissue concentrations achieved with the therapeutic doses only promoted the growth of ovarian cancers. INTERPRETATION: Our results indicate that treatment of ovarian cancer with MF and P4 may induce similar adverse agonistic effects in the absence of classical nuclear PGRs in ovarian cancer. The blockage of PGRMC1 activity may provide a novel treatment strategy for ovarian cancer. FUND: This work was supported by grants from the National Science Centre, Poland (2013/09/N/NZ5/01831 to DP-T; 2012/05/B/NZ5/01867 to MC), Academy of Finland (254366 to NAR), Moikoinen Cancer Research Foundation (to NAR) and EU PARP Cluster grant (UDA-POIG.05.01.00-005/12-00/NCREMFP to SW).


Assuntos
Antineoplásicos Hormonais/farmacologia , Mifepristona/farmacologia , Animais , Antineoplásicos Hormonais/administração & dosagem , Antineoplásicos Hormonais/farmacocinética , Biomarcadores , Carcinoma Epitelial do Ovário/genética , Carcinoma Epitelial do Ovário/metabolismo , Carcinoma Epitelial do Ovário/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Células Cultivadas , Feminino , Humanos , Imuno-Histoquímica , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Transgênicos , Mifepristona/administração & dosagem , Mifepristona/farmacocinética , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo
5.
Neuroscience ; 411: 255-269, 2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-31163207

RESUMO

Repeated stress induces systemic elevations in glucocorticoid levels. Stress is also associated with alterations in central nervous system astrocytes and oligodendrocytes that involve connexins and myelin proteins. Corticosteroid elevation seems a major factor in stress-induced neuropathology. Changes in astrocyte connexins and myelin components may be important mediators for the neurological effects of corticosteroid elevations. Two primary cell culture models, myelination culture from rat embryonic spinal cord (SC) or cerebral cortex (CC) consisting of neurons and glial cells (oligodendrocytes, microglia and astrocytes), and mixed astrocyte-and-oligodendrocyte culture prepared from postnatal rat CC, were used in this study. Cell cultures were treated with either vehicle, corticosterone (CORT) with or without glucocorticoid receptor antagonist mifepristone, or dexamethasone (DEX) during the period of in vitro myelination. Immunoreactivity of astrocyte connexin 43 (Cx43) and oligodendrocyte myelin basic protein (MBP), or the myelination index (co-localization of MBP and phosphorylated neurofilament) was determined by double immunofluorescent labeling. Oligodendrocyte morphology was evaluated by Sholl analysis. Prolonged exposure to CORT or DEX induced dose-dependent reduction of the myelination index, and of immunostaining for MBP and Cx43 in SC and CC myelination cultures, which was prevented by mifepristone. In glial cultures single CORT or DEX exposure caused shrinkage and simplification of/' MBP- or CNPase-positive oligodendrocyte processes. The results support that concurrent effects of glucocorticoids on myelination and astrocyte Cx43 immunoreactivity are mediated by glucocorticoid receptors and may partially account for the involvement of CNS glia in the pathological effects of prolonged stress.


Assuntos
Conexina 43/metabolismo , Dexametasona/farmacologia , Mifepristona/farmacologia , Bainha de Mielina/efeitos dos fármacos , Neuroglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Animais , Células Cultivadas , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Glucocorticoides/farmacologia , Antagonistas de Hormônios/farmacologia , Bainha de Mielina/metabolismo , Neuroglia/citologia , Neuroglia/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Ratos , Medula Espinal/citologia , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo
6.
J Obstet Gynaecol Res ; 45(8): 1530-1535, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31172644

RESUMO

AIM: The aim of this study was to evaluate the efficacy and safety of mifepristone for cervical ripening and induction of labor and compare the results with dinoprostone gel which is an established agent for labor induction. METHODS: A total of 100 patients were enrolled in a prospective study and assigned to one of two treatment protocols. After the exclusion of 10 patients, there were 46 patients in the mifepristone group and 44 in the dinoprostone group. Outcome was evaluated using the improvement in Bishop score, admission delivery interval, duration between induction and the onset of active phase of labor and the mode of delivery. RESULTS: The baseline demographics in the two groups were comparable. The improvement in Bishop's score at first post-intervention assessment was significantly better in dinoprostone group. Duration between instillation and active phase assessment was significantly lesser in dinoprostone group while the admission delivery interval was lesser in mifepristone group. There was no difference in mode of delivery between the two groups. CONCLUSION: The results of the study suggest that oral administration of 200 mg mifepristone in term patients is an effective method of labor induction; and is more convenient and equally safe as compared to intravaginal instillation of dinoprostone.


Assuntos
Dinoprostona/farmacologia , Trabalho de Parto Induzido/métodos , Mifepristona/farmacologia , Ocitócicos/farmacologia , Adulto , Dinoprostona/administração & dosagem , Dinoprostona/efeitos adversos , Feminino , Humanos , Trabalho de Parto Induzido/efeitos adversos , Mifepristona/administração & dosagem , Mifepristona/efeitos adversos , Ocitócicos/administração & dosagem , Ocitócicos/efeitos adversos , Gravidez , Estudos Prospectivos , Resultado do Tratamento , Adulto Jovem
7.
Oxid Med Cell Longev ; 2019: 9752698, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31089421

RESUMO

Aims: Insulin and glucocorticoids play crucial roles in skeletal muscle protein turnover. Fast-twitch glycolytic fibres are more susceptible to atrophy than slow-twitch oxidative fibres. Based on accumulating evidence, hydrogen sulfide (H2S) is a physiological mediator of this process. The regulatory effect of H2S on protein synthesis in fast-twitch fibres was evaluated. Results: A NaHS (sodium hydrosulfide) injection simultaneously increased the diameter of M. pectoralis major (i.e., fast-twitch glycolytic fibres) and activated the mammalian target of the rapamycin (mTOR)/p70S6 kinase (p70S6K) pathway. Dexamethasone (DEX) inhibited protein synthesis, downregulated mTOR and p70S6K phosphorylation, and suppressed the expression of the cystathionine γ-lyase (CSE) protein in myoblasts. The precursor of H2S, L-cysteine, completely abolished the inhibitory effects of DEX. The CSE inhibitor DL-propargylglycine (PAG) completely abrogated the effects of RU486 on blocking the suppressive effects of DEX. The H2S donor NaHS increased the H2S concentrations and abrogated the inhibitory effects of DEX on protein synthesis. Insulin increased protein synthesis and upregulated CSE expression. However, PAG abrogated the stimulatory effects of insulin on protein synthesis and the activity of the mTOR/p70S6K pathway. Innovation: These results demonstrated that CSE/H2S regulated protein synthesis in fast-twitch muscle fibres, and glucocorticoids and insulin regulated protein synthesis in an endogenous CSE/H2S system-dependent manner. Conclusions: The results from the present study suggest that the endogenous CSE/H2S system regulates fast-twitch glycolytic muscle degeneration and regeneration.


Assuntos
Cistationina gama-Liase/metabolismo , Glucocorticoides/farmacologia , Sulfeto de Hidrogênio/metabolismo , Insulina/farmacologia , Proteínas Musculares/biossíntese , Biossíntese de Proteínas/efeitos dos fármacos , Alquinos/farmacologia , Animais , Galinhas , Cisteína/farmacologia , Dexametasona/farmacologia , Glicina/análogos & derivados , Glicina/farmacologia , Injeções Intraperitoneais , Mifepristona/farmacologia , Mioblastos/efeitos dos fármacos , Mioblastos/metabolismo , Fosforilação/efeitos dos fármacos , Receptores de Glucocorticoides/metabolismo , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo
8.
Br Poult Sci ; 60(4): 395-403, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31132872

RESUMO

1. In this study, geese (Anas cygnoides) embryonic pituitary cells were cultured in vitro to determine if glucocorticoids could induce growth hormone (GH) expression and to investigate the molecular mechanisms involved in this process. 2. On embryonic day 15 (e15) and e20 the pituitary cells were treated with corticosterone (CORT), membrane impermeable bovine serum albumin-conjugate corticosterone (CORT-BSA), dexamethasone (DEX), and a glucocorticoid receptor (GR) antagonist (RU486) to detect responsiveness of somatotrophs to glucocorticoids. 3. Treatment with CORT, CORT-BSA, and DEX for as little as 6 h increased the percentage of GH-positive cells (P < 0.01) and increased GH mRNA expression (P < 0.01) in e15 goose pituitary cells compared to the control. CORT significantly increased the level of GH protein secreted from cultured e15 goose embryonic pituitary cells, and CORT-BSA increased GH secretion from e20 goose embryonic pituitary cells. 4. A significant increase was observed in the glucocorticoid receptor in GR transcription levels (P < 0.01) with CORT, CORT-BSA, and DEX treatment. Furthermore, the CORT-stimulated GH mRNA expression was completely negated by pre-treatment with RU486. 5. These findings demonstrate that glucocorticoids can stimulate somatotroph differentiation in vitro, as characterised by enhanced GH protein secretion andmRNA expression in cultured geese embryonic pituitary cells. The membrane GR was involved in pituitary somatotroph differentiation induced by glucocorticoids during the embryonic development of geese.


Assuntos
Diferenciação Celular/fisiologia , Corticosterona/farmacologia , Cortisona/farmacologia , Dexametasona/farmacologia , Gansos/fisiologia , Receptores de Glucocorticoides/metabolismo , Soroalbumina Bovina/farmacologia , Somatotrofos/fisiologia , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Células Cultivadas , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/fisiologia , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/fisiologia , Gansos/genética , Antagonistas de Hormônios/farmacologia , Mifepristona/farmacologia , Hipófise/fisiologia , RNA Mensageiro/metabolismo
9.
BMC Cancer ; 19(1): 376, 2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-31014286

RESUMO

BACKGROUND: Previous work in our laboratory demonstrated that antiprogestin mifepristone impairs the growth and adhesion of highly metastatic cancer cells, and causes changes in their cellular morphology. In this study, we further assess the anti-metastatic properties of mifepristone, by studying whether cytostatic doses of the drug can inhibit the migration and invasion of various cancer cell lines using a double fluorescence cytochemical labeling approach. METHODS: Cell lines representing cancers of the ovary (SKOV-3), breast (MDA-MB-231), glia (U87MG), or prostate (LNCaP) were treated with cytostatic concentrations of mifepristone. Wound healing and Boyden chamber assays were utilized to study cellular migration. To study cellular invasion, the Boyden chamber assay was prepared by adding a layer of extracellular matrix over the polycarbonate membrane. We enhanced the assays with the addition of double fluorescence cytochemical staining for fibrillar actin (F-actin) and DNA to observe the patterns of cytoskeletal distribution and nuclear positioning while cells migrate and invade. RESULTS: When exposed to cytostatic concentrations of mifepristone, all cancer cells lines demonstrated a decrease in both migration and invasion capacities measured using standard approaches. Double fluorescence cytochemical labeling validated that mifepristone-treated cancer cells exhibit reduced migration and invasion, and allowed to unveil a distinct migration pattern among the different cell lines, different arrays of nuclear localization during migration, and apparent redistribution of F-actin to the nucleus. CONCLUSION: This study reports that antiprogestin mifepristone inhibits migration and invasion of highly metastatic cancer cell lines, and that double fluorescence cytochemical labeling increases the value of well-known approaches to study cell movement.


Assuntos
Movimento Celular , Proliferação de Células , Fluorescência , Antagonistas de Hormônios/farmacologia , Mifepristona/farmacologia , Neoplasias/patologia , Humanos , Invasividade Neoplásica , Neoplasias/tratamento farmacológico , Células Tumorais Cultivadas , Cicatrização
10.
Reprod Biol ; 19(1): 14-21, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30852242

RESUMO

Heart and neural crest derivatives-expressed transcript 2 (HAND2) is a key transcription factor in progestin-induced decidualization of human endometrial stromal cells (ESCs). In the mouse, HAND2 plays an important role in uterine receptivity by suppressing several fibroblast growth factors (FGFs). However, the regulation of FGF family members by progestin-induced HAND2 and the role of FGF in vascular regeneration in the endometrium remains poorly understood. To investigate these molecular mechanisms, primary human ESCs were cultured with estradiol (E2), medroxyprogesterone acetate (MPA), progesterone receptor (PR) antagonist RU486, HAND2-specific small interfering RNA (siRNA), and recombinant FGF. The expression levels of FGF family members, HAND2, angiopoietin (ANGPT), and vascular endothelial growth factor (VEGF) were assessed by real-time PCR and ELISA. Out of six FGF genes known to be expressed in the human endometrium, only one, FGF9, was significantly downregulated in human ESCs after 3 days of progestin treatment. E2 + MPA attenuated the mRNA and protein levels of FGF9 during decidualization of ESCs, and this effect was blocked by RU486. Silencing of HAND2 significantly increased FGF9 expression in ESCs treated with E2 + MPA. Moreover, FGF9 activated FGF receptor in human ESCs, triggering ANGPT2 production, which resulted in enhancement of the ANGPT2/ANGPT1 protein ratio. Taken together, progestin-PR signaling and its target HAND2 play an essential role in FGF9 suppression in the human endometrium. In addition, progestin-induced HAND2 inhibits ANGPT2 production by suppressing FGF9 in ESCs. These results suggest that HAND2 may contribute to endometrial vascular maturation by regulating FGF9 during decidualization.


Assuntos
Angiopoietina-2/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Fator 9 de Crescimento de Fibroblastos/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Progestinas/farmacologia , Células Estromais/efeitos dos fármacos , Angiopoietina-2/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Endométrio/citologia , Estradiol/farmacologia , Feminino , Fator 9 de Crescimento de Fibroblastos/genética , Humanos , Luteolíticos/farmacologia , Acetato de Medroxiprogesterona/farmacologia , Mifepristona/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno , Células Estromais/metabolismo
11.
Cells ; 8(3)2019 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-30841579

RESUMO

Epigenetic regulation has been considered an important mechanism for influencing stem cell differentiation. In particular, histone deacetylases (HDACs) have been shown to play a role in the osteoblast differentiation of mesenchymal stem cells (MSCs). In this study, the effect of the HDAC inhibitor, valproic acid (VPA), on bone formation in vivo by MSCs was determined. Surprisingly, VPA treatment, unlike other HDAC inhibitors, produced a well-organized lamellar bone tissue when MSCs⁻collagen sponge constructs were implanted subcutaneously into nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice, although a decrease of osteocalcin (OC) expression was observed. Consequently, we decided to investigate the molecular mechanisms by which VPA exerts such effects on MSCs. We identified the glucocorticoid receptor (GR) as being responsible for that downregulation, and suggested a correlation between GR and HDAC2 inhibition after VPA treatment, as evidenced by HDAC2 knockdown. Furthermore, using co-immunoprecipitation analysis, we showed for the first time in the cytoplasm, binding between GR and HDAC2. Additionally, chromatin immunoprecipitation (ChIP) assays confirmed the role of GR in OC downregulation, showing recruitment of GR to the nGRE element in the OC promoter. In conclusion, our results highlight the existence of a cross-talk between GR and HDAC2, providing a mechanistic explanation for the influence of the HDAC inhibitor (namely VPA) on osteogenic differentiation in MSCs. Our findings open new directions in targeted therapies, and offer new insights into the regulation of MSC fate determination.


Assuntos
Citoplasma/metabolismo , Histona Desacetilase 2/metabolismo , Células-Tronco Mesenquimais/metabolismo , Osteocalcina/metabolismo , Receptores de Glucocorticoides/metabolismo , Ácido Valproico/farmacologia , Adulto , Biomarcadores/metabolismo , Citoplasma/efeitos dos fármacos , Polpa Dentária/citologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Humanos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Mifepristona/farmacologia , Osteocalcina/genética , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Implantação de Prótese , Ligação Proteica/efeitos dos fármacos , Elementos de Resposta/genética , Adulto Jovem
12.
Diabetes ; 68(5): 918-931, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30833469

RESUMO

Low 25-hydroxyvitamin D levels correlate with the prevalence of diabetes; however, the mechanisms remain uncertain. Here, we show that nutritional deprivation-responsive mechanisms regulate vitamin D metabolism. Both fasting and diabetes suppressed hepatic cytochrome P450 (CYP) 2R1, the main vitamin D 25-hydroxylase responsible for the first bioactivation step. Overexpression of coactivator peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α), induced physiologically by fasting and pathologically in diabetes, resulted in dramatic downregulation of CYP2R1 in mouse hepatocytes in an estrogen-related receptor α (ERRα)-dependent manner. However, PGC-1α knockout did not prevent fasting-induced suppression of CYP2R1 in the liver, indicating that additional factors contribute to the CYP2R1 repression. Furthermore, glucocorticoid receptor (GR) activation repressed the liver CYP2R1, suggesting GR involvement in the regulation of CYP2R1. GR antagonist mifepristone partially prevented CYP2R1 repression during fasting, suggesting that glucocorticoids and GR contribute to the CYP2R1 repression during fasting. Moreover, fasting upregulated the vitamin D catabolizing CYP24A1 in the kidney through the PGC-1α-ERRα pathway. Our study uncovers a molecular mechanism for vitamin D deficiency in diabetes and reveals a novel negative feedback mechanism that controls crosstalk between energy homeostasis and the vitamin D pathway.


Assuntos
Diabetes Mellitus/metabolismo , Jejum/sangue , Fatores de Transcrição/sangue , Fatores de Transcrição/metabolismo , Deficiência de Vitamina D/metabolismo , Vitamina D/sangue , Vitamina D/metabolismo , Animais , Colestanotriol 26-Mono-Oxigenase/metabolismo , Diabetes Mellitus/sangue , Jejum/fisiologia , Fígado/metabolismo , Camundongos , Mifepristona/farmacologia , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Receptores Estrogênicos/metabolismo , Receptores de Glucocorticoides/antagonistas & inibidores , Receptores de Glucocorticoides/metabolismo , Deficiência de Vitamina D/sangue
13.
Mol Cell Endocrinol ; 485: 88-96, 2019 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-30796948

RESUMO

The aim of this study was to analyze the effects of progesterone withdrawal on gene transcription in receptive endometrium by the administration of a single dose of 50 mg of the anti-progesterone receptor mifepristone (MFP) at the time of follicle rupture (FR). Six volunteer ovulatory women were studied, taking endometrial biopsies of three control and three MFP-treated women on days LH+2 (C-LH+2) and LH+7 (T-MFP), respectively. The biopsies were prepared for RNA isolation or histological and immunohistochemistry studies. The genomic data from 14 women (C-LH+7) were included as a historical control. The functional genomic analysis of the differentially expressed genes showed that MFP interfered negatively with the bio-functions decidualization of uterus and implantation of blastocyst and embryo. The results of this study confirm but also give new information on how MFP affects endometrial gene expression when administered at the time of FR and the dose used in emergency contraception.


Assuntos
Endométrio/química , Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes/efeitos dos fármacos , Antagonistas de Hormônios/administração & dosagem , Mifepristona/administração & dosagem , Adulto , Estudos de Casos e Controles , Endométrio/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Ontologia Genética , Genômica/métodos , Humanos , Mifepristona/farmacologia , Ovulação/efeitos dos fármacos , Adulto Jovem
14.
Int J Mol Sci ; 20(2)2019 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-30642045

RESUMO

Endogenous γ-aminobutyric acid (GABA)-dependent activity induces death of developing Purkinje neurons in mouse organotypic cerebellar cultures and the synthetic steroid mifepristone blocks this effect. Here, using brain-derived neurotrophic factor (BDNF) heterozygous mice, we show that BDNF plays no role in immature Purkinje cell death. However, interestingly, BDNF haploinsufficiency impairs neuronal survival induced by mifepristone and GABAA-receptors antagonist (bicuculline) treatments, indicating that the underlying neuroprotective mechanism requires the neurotrophin full expression.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Mifepristona/farmacologia , Fármacos Neuroprotetores/farmacologia , Células de Purkinje/citologia , Ácido gama-Aminobutírico/efeitos adversos , Animais , Apoptose , Bicuculina/farmacologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Sobrevivência Celular , Células Cultivadas , Haploinsuficiência , Camundongos , Células de Purkinje/efeitos dos fármacos , Células de Purkinje/metabolismo
15.
Int J Mol Sci ; 20(2)2019 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-30650648

RESUMO

Phospholipids (PLs) possess the unique ability to contribute to synovial joint lubrication. The aim of our study was to determine for the first time the effect of dexamethasone and some adrenergic and cholinergic agonists on the biosynthesis and release of PLs from human fibroblast-like synoviocytes (FLS). Osteoarthritic human knee FLS were treated with dexamethasone, terbutaline, epinephrine, carbachol, and pilocarpine, or the glucocorticoid receptor antagonist RU 486. Simultaneously PL biosynthesis was determined through the incorporation of stable isotope-labeled precursors into PLs. Radioactive isotope-labeled precursors were used to radiolabel PLs for the subsequent quantification of their release into nutrient media. Lipids were extracted and quantified using electrospray ionization tandem mass spectrometry or liquid scintillation counting. Dexamethasone significantly decreased the biosynthesis of phosphatidylcholine, phosphatidylethanolamine (PE), PE-based plasmalogen, and sphingomyelin. The addition of RU 486 abolished these effects. A release of PLs from FLS into nutrient media was not recognized by any of the tested agents. None of the adrenergic or cholinergic receptor agonists modulated the PL biosynthesis. We demonstrate for the first time an inhibitory effect of dexamethasone on the PL biosynthesis of FLS from human knees. Moreover, our study indicates that the PL metabolism of synovial joints and lungs are differently regulated.


Assuntos
Agonistas Adrenérgicos/farmacologia , Agonistas Colinérgicos/farmacologia , Dexametasona/farmacologia , Osteoartrite/patologia , Fosfolipídeos/metabolismo , Receptores Adrenérgicos/metabolismo , Receptores Colinérgicos/metabolismo , Sinoviócitos/patologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mifepristona/farmacologia , Modelos Biológicos , Osteoartrite/metabolismo , Fosfolipídeos/biossíntese , Sinoviócitos/efeitos dos fármacos
16.
Behav Brain Res ; 360: 146-157, 2019 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-30500430

RESUMO

Somatic and motivational symptoms accompanying opiate withdrawal are considered one of the major reasons for relapse to opiate-seeking and opiate-taking behaviors. These symptoms are accompanied by the activation of stress-related processes including hypothalamic-pituitary-adrenal axis activity and noradrenergic (NA) signaling. In particular, the NA system plays an important role in the expression of somatic signs of opiate withdrawal, whereas glucocorticoid (GR) and mineralocorticoid receptors (MR) are activated during opiate abstinence. The purpose of our study was to examine the roles of α1-, α2-, and ß-adrenoceptors (ARs) as well as GR and MR, in the formation and expression of physiological and motivational symptoms of morphine withdrawal. We showed that systemic pretreatment with the selective α1-AR antagonist prazosin (0-1 mg/kg), the selective α2-AR antagonist RX821002 (0-2 mg/kg), the selective ß-adrenergic antagonist, propranolol (0-10 mg/kg), or the selective MR antagonist spironolactone (0-50 mg/kg), but not the selective GR antagonist mifepristone (0-40 mg/kg), decreased somatic symptoms of naloxone-precipitated morphine withdrawal in mice chronically treated with morphine. In contrast, only propranolol pretreatment attenuated the dysphoric affective state accompanying naloxone-precipitated morphine withdrawal as assessed in the conditioned place aversion (N-CPA) paradigm. Together, our results demonstrate the important roles of noradrenergic receptors in the modulation of somatic, but not motivational/affective, symptoms of morphine withdrawal. In addition MR but not GR regulates the expression of only somatic symptoms of morphine withdrawal.


Assuntos
Transtornos do Humor/etiologia , Morfina/toxicidade , Receptores Adrenérgicos/metabolismo , Receptores de Esteroides/metabolismo , Distúrbios Somatossensoriais/etiologia , Síndrome de Abstinência a Substâncias/complicações , Antagonistas de Receptores Adrenérgicos alfa 1/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Antagonistas de Hormônios/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mifepristona/farmacologia , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Entorpecentes , Uso Off-Label , Prazosina/farmacologia , Propranolol/farmacologia , Síndrome de Abstinência a Substâncias/psicologia
17.
Acta Obstet Gynecol Scand ; 98(2): 250-261, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30325501

RESUMO

INTRODUCTION: Women with hereditary mutation in breast cancer-associated genes (BRCA1-/2- ) have a higher lifetime risk of developing ovarian cancer. Here, we aimed to investigate the effect of mifepristone, a selective progesterone receptor modulator of ovarian mesenchymal stem/stromal cells (MSC) from BRCA1-/2- carriers. MATERIAL AND METHODS: Ovarian BRCA1-/2- MSC were positively selected using the markers CD90, CD73 and CD105 from nine healthy women. The effect of dose response and combination treatment with mifepristone and analogs of progesterone- or glucocorticoid-receptors were investigated on BRCA1-/2- MSC in vitro using a panel of markers for proliferation (ki67, BrdU, CDK2, p21CIP ), apoptosis (BAX, BCL2, CASPASE3), tumor suppression (TP53, PTEN) and cell survival (PI3KCA, MAPK3, mTOR). RESULTS: The dose response with mifepristone treatment suggested an optimal effect with 10 µm mifepristone, exhibiting >90% viability and significantly reducing growth signaling markers (TP53 and MAPK3). Furthermore, combined treatment with progesterone plus mifepristone (PG+MIFE) gave an enhanced anti-proliferative effect in comparison with hydrocortisone plus mifepristone (HC+MIFE) by significantly reducing markers of proliferation (BrdU+ and Ki67 expression) and tumor suppressors (PTEN, TP53), and increasing the percentage of pro-apoptotic cells. Consequently, accumulation of p21CIP together with reduced levels of CDK2 confirms growth inhibition by reversibly arresting cell-cycle progression at the G1-S phase, not by inducing apoptosis. CONCLUSIONS: Our study showed an anti-proliferative effect on ovarian BRCA1-/2- MSC on in vitro combined treatment with mifepristone and progesterone. These findings suggest that mifepristone or other selective progesterone receptor modulators could be developed as a preventive treatment and postpone early use of prophylactic salpingo-oophorectomy as well as reduce the risk of ovarian cancer.


Assuntos
Proteína BRCA1/genética , Proteína BRCA2/genética , Proliferação de Células/efeitos dos fármacos , Células-Tronco Mesenquimais , Mifepristona/farmacologia , Ovário , Células Estromais , Sobrevivência Celular/efeitos dos fármacos , Feminino , Antagonistas de Hormônios/farmacocinética , Antagonistas de Hormônios/farmacologia , Humanos , Células-Tronco Mesenquimais/patologia , Células-Tronco Mesenquimais/fisiologia , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Ovário/efeitos dos fármacos , Ovário/metabolismo , Ovário/patologia , Receptores de Glucocorticoides/metabolismo , Receptores de Progesterona/metabolismo , Células Estromais/patologia , Células Estromais/fisiologia , Células Tumorais Cultivadas
18.
Laryngoscope ; 129(5): E187-E193, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30325506

RESUMO

OBJECTIVES/HYPOTHESIS: Direct glucocorticoid (GC) injection for vocal fold (VF) scarring has evolved as a therapeutic strategy, but the mechanisms underlying the antifibrotic effects remain unclear. GCs act via the glucocorticoid receptor (GR), which is phosphorylated at multiple serine residues in a hormone-dependent manner to affect bioactivity. We hypothesize that GCs regulate SMAD signaling via GR phosphorylation in vocal fold fibroblasts (VFFs). STUDY DESIGN: In vitro. METHODS: Human VFFs were treated with dexamethasone (DM; 10-5 -10-7 M) ± transforming growth factor (TGF)-ß1 (10 ng/mL). RU486 (10-6 M) was employed to isolate the regulatory effects of GR. Total GR, Ser211 , and Ser203 phosphorylation was examined via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunocytochemistry. Quantitative polymerase chain reaction was employed to determine GR-mediated effects of DM on genes related to fibrosis. RESULTS: Total GR and Ser211 phosphorylation was observed predominantly in the nucleus 1 hour after DM administration. DM decreased total GR expression, but Ser203 and Ser211 phosphorylation increased. RU486 limited the effects of DM. SMAD3 and SMAD7 mRNA expression significantly decreased 4 hours after DM administration (P < 0.05); this response was negated by RU486. COL1A1 remained unchanged, and ACTA2 significantly increased following 24 hours of DM treatment (P < 0.05). CONCLUSION: DM regulated TGF-ß1 signaling via altered SMAD3 and SMAD7 expression. This response was associated with altered GR phosphorylation. These findings provide insight into the mechanisms of steroidal effects on vocal fold repair; ultimately, we seek to enhance therapeutic strategies for these challenging patients. LEVEL OF EVIDENCE: NA Laryngoscope, 129:E187-E193, 2019.


Assuntos
Receptores de Glucocorticoides/metabolismo , Prega Vocal/efeitos dos fármacos , Prega Vocal/metabolismo , Actinas/metabolismo , Linhagem Celular , Dexametasona/farmacologia , Eletroforese em Gel de Poliacrilamida , Fibroblastos/metabolismo , Humanos , Imuno-Histoquímica , Mifepristona/farmacologia , Fosforilação , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Transdução de Sinais , Proteína Smad3/metabolismo , Proteína Smad7/metabolismo , Fator de Crescimento Transformador beta/metabolismo
19.
Toxicol In Vitro ; 54: 33-40, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30217652

RESUMO

Etoposide is a well-known and widely used anticancer drug that displays several side effects. In addition, tumors often acquire resistance to this drug. Our aim is to develop a combination therapy that would augment toxicity of etoposide in malignant cells. Based on literature and our experiments, we selected mifepristone (RU486) as a potential supporting molecule that is able to enhance etoposide toxicity against cancer cells. All experiments were performed with Hep G2 cells, a well-known and described human hepatocellular carcinoma cell line. By using xCELLigence system, we demonstrated that mifepristone enhances toxicity of etoposide in a dose dependent manner with concomitant caspase-3 activity. We evaluated upregulation of Bax because mifepristone was demonstrated to modulate proapoptotic Bax protein expression. Our data show only weak and not statistically significant increase of Bax expression. On the other hand, we show that mifepristone increases etoposide toxicity via inhibition of ABC transporters, coupled with significant increase of intracellular etoposide concentration. In conclusion, we demonstrate that mifepristone has a synergistic effect with etoposide treatment in the Hep G2 cells and that the effect is related to ABC transporters inhibition.


Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/antagonistas & inibidores , Antineoplásicos Fitogênicos/farmacologia , Etoposídeo/farmacologia , Mifepristona/farmacologia , Proteínas de Neoplasias/antagonistas & inibidores , Subfamília B de Transportador de Cassetes de Ligação de ATP/antagonistas & inibidores , Transporte Biológico/efeitos dos fármacos , Caspase 3/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sinergismo Farmacológico , Células Hep G2 , Humanos , Células K562 , Proteína X Associada a bcl-2/metabolismo
20.
Cell Mol Neurobiol ; 39(4): 503-522, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30173378

RESUMO

Psychotic depression is characterized by elevated circulating cortisol, and high daily doses of the glucocorticoid/progesterone antagonist mifepristone for 1 week are required for significant improvement. Using a rodent model, we find that such high doses of mifepristone are needed because the antagonist is rapidly degraded and poorly penetrates the blood-brain barrier, but seems to facilitate the entry of cortisol. We also report that in male C57BL/6J mice, after a 7-day treatment with a high dose of mifepristone, basal blood corticosterone levels were similar to that of vehicle controls. This is surprising because after the first mifepristone challenge, corticosterone remained elevated for about 16 h, and then decreased towards vehicle control levels at 24 h. At that time, stress-induced corticosterone levels of the 1xMIF were sevenfold higher than the 7xMIF group, the latter response being twofold lower than controls. The 1xMIF mice showed behavioral hyperactivity during exploration of the circular hole board, while the 7xMIF mice rather engaged in serial search patterns. To explain this rapid reset of corticosterone secretion upon recurrent mifepristone administration, we suggest the following: (i) A rebound glucocorticoid feedback after cessation of mifepristone treatment. (ii) Glucocorticoid agonism in transrepression and recruitment of cell-specific coregulator cocktails. (iii) A more prominent role of brain MR function in control of stress circuit activity. An overview table of neuroendocrine MIF effects is provided. The data are of interest for understanding the mechanistic underpinning of stress system reset as treatment strategy for stress-related diseases.


Assuntos
Mifepristona/farmacologia , Estresse Fisiológico/efeitos dos fármacos , Animais , Comportamento Animal/efeitos dos fármacos , Transporte Biológico/efeitos dos fármacos , Ritmo Circadiano/efeitos dos fármacos , Corticosterona/sangue , Corticosterona/metabolismo , Hormônio Liberador da Corticotropina/genética , Hormônio Liberador da Corticotropina/metabolismo , Humanos , Hidrocortisona/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Mifepristona/administração & dosagem , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Glucocorticoides/antagonistas & inibidores , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/metabolismo , Receptores de Mineralocorticoides/genética , Receptores de Mineralocorticoides/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA