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1.
Ecotoxicol Environ Saf ; 186: 109776, 2019 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-31606647

RESUMO

In the present study, to evaluate neurobehavioral toxicity and the thyroid-disrupting effects of environmental levels of triphenyltin (TPT), the zebrafish larvae were exposed to 1, 10 and 100 ng/l TPT. In the neurobehavioral assay, increased levels of dopamine and serotonin, decreased content of nitric oxide, inhibited activities of acetylcholinesterase and monoamine oxidase were observed in the whole body of zebrafish larvae after TPT treatment, as well as the serious abnormal non-reproductive behavior. Moreover, the whole-body the T4 levels were markedly decreased significantly, whereas T3 levels were not significantly changed under TPT stress. In addition, TPT exposure significantly changed the expression levels of genes related to thyroid system, including corticotropin-releasing hormone gene crh, thyroid-stimulating hormone gene tshß, thyroglobulin gene tg, sodium/iodide symporter gene nis, thyroid hormone nuclear receptor trα, isoform trß, types I deiodinase gene dio1and types II deiodinase gene dio2. The regulated responsiveness of thyroid hormone and related genes expression levels suggested that TPT could induce the thyroid disrupting effects in zebrafish larvae. Therefore, our results provide new aspects of TPT as an endocrine disrupting chemical.


Assuntos
Comportamento Animal/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Compostos Orgânicos de Estanho/toxicidade , Glândula Tireoide/efeitos dos fármacos , Peixe-Zebra/metabolismo , Animais , Hormônio Liberador da Corticotropina/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Iodeto Peroxidase/genética , Larva/efeitos dos fármacos , Larva/metabolismo , Glândula Tireoide/metabolismo , Hormônios Tireóideos/metabolismo , Tireotropina/genética , Proteínas de Peixe-Zebra/metabolismo
2.
Anal Biochem ; 570: 56-61, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30768924

RESUMO

One of the most used formats in inmuno-polymerase chain reaction (IPCR) is known as "Universal" IPCR (signal-generating complexes is based on conjugates of biotinylated DNA, biotinylated IgG and avidin). In the present study, we evaluated the utility of using mono- and bi-biotinylated DNA probes, pre-self-assembled DNA-neutravidin complex, blocking step and glutaraldehyde pretreatment of standard PCR tubes to improve the analytical performance of the hTSH-IPCR assay. The use of pre-self-assembled mono-biotinylated DNA-neutravidin complex enhances both the sensitivity and the reproducibility of the hTSH-IPCR assay, even without blocking step: hTSH-IPCR assay showed an improved limit of detection (LOD: 0.01 µIU/ml), calibration sensitivity (SEN: 2.4) and analytic sensitivity (γ: 9 µIU/ml-1) in comparison with both a self-made ELISA and a commercial one.


Assuntos
Reação em Cadeia da Polimerase/métodos , Tireotropina/análise , Biotinilação , Sondas de DNA/química , Sondas de DNA/metabolismo , Humanos , Imunoensaio , Limite de Detecção , Reação em Cadeia da Polimerase/instrumentação , Reprodutibilidade dos Testes , Tireotropina/genética
3.
JAMA Cardiol ; 4(2): 136-143, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30673079

RESUMO

Importance: Thyroid hormone levels are tightly regulated through feedback inhibition by thyrotropin, produced by the pituitary gland. Hyperthyroidism is overwhelmingly due to thyroid disorders and is well recognized to contribute to a wide spectrum of cardiovascular morbidity, particularly the increasingly common arrhythmia atrial fibrillation (AF). Objective: To determine the association between genetically determined thyrotropin levels and AF. Design, Setting, and Participants: This phenome-wide association study scanned 1318 phenotypes associated with a polygenic predictor of thyrotropin levels identified by a previously published genome-wide association study that included participants of European ancestry. North American individuals of European ancestry with longitudinal electronic health records were analyzed from May 2008 to November 2016. Analysis began March 2018. Main Outcomes and Measures: Clinical diagnoses associated with a polygenic predictor of thyrotropin levels. Exposures: Genetically determined thyrotropin levels. Results: Of 37 154 individuals, 19 330 (52%) were men. The thyrotropin polygenic predictor was positively associated with hypothyroidism (odds ratio [OR], 1.10; 95% CI, 1.07-1.14; P = 5 × 10-11) and inversely associated with diagnoses related to hyperthyroidism (OR, 0.64; 95% CI, 0.54-0.74; P = 2 × 10-8 for toxic multinodular goiter). Among nonthyroid associations, the top association was AF/flutter (OR, 0.93; 95% CI, 0.9-0.95; P = 9 × 10-7). When the analyses were repeated excluding 9801 individuals with any diagnoses of a thyroid-related disease, the AF association persisted (OR, 0.91; 95% CI, 0.88-0.95; P = 2.9 × 10-6). To replicate this association, we conducted an inverse-variance weighted average meta-analysis using AF single-nucleotide variant weights from a genome-wide association study of 17 931 AF cases and 115 142 controls. As in the discovery analyses, each SD increase in predicted thyrotropin was associated with a decreased risk of AF (OR, 0.86; 95% CI, 0.79-0.93; P = 4.7 × 10-4). In a set of AF cases (n = 745) and controls (n = 1680) older than 55 years, directly measured thyrotropin levels that fell within the normal range were inversely associated with AF risk (OR, 0.91; 95% CI, 0.83-0.99; P = .04). Conclusions and Relevance: This study suggests a role for genetically determined variation in thyroid function within a physiologically accepted normal range as a risk factor for AF. The clinical decision to treat subclinical thyroid disease should incorporate the risk for AF as antithyroid medications to treat hyperthyroidism may reduce AF risk and thyroid hormone replacement for hypothyroidism may increase AF risk.


Assuntos
Fibrilação Atrial/genética , Hipertireoidismo/genética , Hipotireoidismo/genética , Glândula Tireoide/metabolismo , Tireotropina/genética , Idoso , Análise de Variância , Fibrilação Atrial/epidemiologia , Fibrilação Atrial/etiologia , Grupo com Ancestrais do Continente Europeu/genética , Feminino , Estudo de Associação Genômica Ampla , Humanos , Hipertireoidismo/complicações , Hipertireoidismo/patologia , Hipotireoidismo/complicações , Masculino , Pessoa de Meia-Idade , Fenótipo , Polimorfismo de Nucleotídeo Único , Valor Preditivo dos Testes , Fatores de Risco , Testes de Função Tireóidea/métodos , Glândula Tireoide/fisiopatologia
4.
JAMA Cardiol ; 4(2): 144-152, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30673084

RESUMO

Importance: Increased free thyroxine (FT4) and decreased thyrotropin are associated with increased risk of atrial fibrillation (AF) in observational studies, but direct involvement is unclear. Objective: To evaluate the potential direct involvement of thyroid traits on AF. Design, Setting, and Participants: Study-level mendelian randomization (MR) included 11 studies, and summary-level MR included 55 114 AF cases and 482 295 referents, all of European ancestry. Exposures: Genomewide significant variants were used as instruments for standardized FT4 and thyrotropin levels within the reference range, standardized triiodothyronine (FT3):FT4 ratio, hypothyroidism, standardized thyroid peroxidase antibody levels, and hyperthyroidism. Mendelian randomization used genetic risk scores in study-level analysis or individual single-nucleotide polymorphisms in 2-sample MR for the summary-level data. Main Outcomes and Measures: Prevalent and incident AF. Results: The study-level analysis included 7679 individuals with AF and 49 233 referents (mean age [standard error], 62 [3] years; 15 859 men [29.7%]). In study-level random-effects meta-analysis, the pooled hazard ratio of FT4 levels (nanograms per deciliter) for incident AF was 1.55 (95% CI, 1.09-2.20; P = .02; I2 = 76%) and the pooled odds ratio (OR) for prevalent AF was 2.80 (95% CI, 1.41-5.54; P = .003; I2 = 64%) in multivariable-adjusted analyses. The FT4 genetic risk score was associated with an increase in FT4 by 0.082 SD (standard error, 0.007; P < .001) but not with incident AF (risk ratio, 0.84; 95% CI, 0.62-1.14; P = .27) or prevalent AF (OR, 1.32; 95% CI, 0.64-2.73; P = .46). Similarly, in summary-level inverse-variance weighted random-effects MR, gene-based FT4 within the reference range was not associated with AF (OR, 1.01; 95% CI, 0.89-1.14; P = .88). However, gene-based increased FT3:FT4 ratio, increased thyrotropin within the reference range, and hypothyroidism were associated with AF with inverse-variance weighted random-effects OR of 1.33 (95% CI, 1.08-1.63; P = .006), 0.88 (95% CI, 0.84-0.92; P < .001), and 0.94 (95% CI, 0.90-0.99; P = .009), respectively, and robust to tests of horizontal pleiotropy. However, the subset of hypothyroidism single-nucleotide polymorphisms involved in autoimmunity and thyroid peroxidase antibodies levels were not associated with AF. Gene-based hyperthyroidism was associated with AF with MR-Egger OR of 1.31 (95% CI, 1.05-1.63; P = .02) with evidence of horizontal pleiotropy (P = .045). Conclusions and Relevance: Genetically increased FT3:FT4 ratio and hyperthyroidism, but not FT4 within the reference range, were associated with increased AF, and increased thyrotropin within the reference range and hypothyroidism were associated with decreased AF, supporting a pathway involving the pituitary-thyroid-cardiac axis.


Assuntos
Fibrilação Atrial/genética , Análise da Randomização Mendeliana/métodos , Glândula Tireoide/metabolismo , Glândula Tireoide/patologia , Idoso , Fibrilação Atrial/diagnóstico , Fibrilação Atrial/epidemiologia , Grupo com Ancestrais do Continente Europeu/genética , Feminino , Estudo de Associação Genômica Ampla/métodos , Humanos , Hipertireoidismo/sangue , Hipertireoidismo/epidemiologia , Hipertireoidismo/genética , Hipotireoidismo/sangue , Hipotireoidismo/epidemiologia , Hipotireoidismo/genética , Iodeto Peroxidase/imunologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Fatores de Risco , Testes de Função Tireóidea/métodos , Glândula Tireoide/fisiopatologia , Tireotropina/sangue , Tireotropina/genética , Tiroxina/sangue , Tiroxina/genética , Tri-Iodotironina/sangue , Tri-Iodotironina/genética
5.
Clin Cancer Res ; 25(1): 414-425, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30314969

RESUMO

PURPOSE: Thyroid-stimulating hormone (TSH) suppression is widely used to treat well-differentiated thyroid cancer, whereas its role in poorly differentiated thyroid cancer (PDTC) is undetermined. Besides thyrocytes, TSH also binds to stromal cells, comprising tumor microenvironments. This study aimed to investigate the effects of TSH on tumor microenvironments in PDTC. EXPERIMENTAL DESIGN: An ectopic tumor model using PDTC cells (BHP10-3SCp and FRO), which exhibit TSH/cAMP-independent cell growth, was treated with TSH. IHC was performed using tissue microarrays from 13 PDTCs. RESULTS: TSH treatment significantly enhanced tumor growth of PDTCs with increased vascularity but not that of breast cancer cells, suggesting this effect is unique to thyroid cancer cells, not stromal cells. TSH significantly upregulated VEGF-A and CXCL8 expressions in BHP10-3SCp cells via AKT and ERK signaling, resulting in higher concentrations of VEGF-A and CXCL8 in conditioned medium of TSH-treated BHP10-3SCp cells (TSH-CM) compared with controls. TSH-CM treatment enhanced tube formation potentials of endothelial cells, and blocking VEGF and/or CXCL8 reduced them. Blocking VEGF and/or CXCL8 also reduced TSH-dependent tumor growth with reduced tumor vasculature in vivo. TSH-treated tumors showed increased macrophage densities, and macrophage inhibition reduced TSH-dependent tumor growth in vivo. In human PDTCs, preoperative TSH levels were positively associated with VEGF-A and tumor size, and the expression of VEGF-A was positively correlated with CD31, CD163, and CXCL8, and their clinical poor prognosis. CONCLUSIONS: Aberrant TSH receptor signaling modulates tumor angiogenesis by stimulating VEGF-A and CXCL8 secretion from PDTC cells and enhances tumor growth; thus, TSH suppression is beneficial for treating PDTCs.


Assuntos
Interleucina-8/genética , Neovascularização Patológica/genética , Receptores da Tireotropina/genética , Neoplasias da Glândula Tireoide/genética , Fator A de Crescimento do Endotélio Vascular/genética , Inibidores da Angiogênese/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , Humanos , Macrófagos/metabolismo , Macrófagos/patologia , Camundongos , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/patologia , Receptores da Tireotropina/antagonistas & inibidores , Neoplasias da Glândula Tireoide/tratamento farmacológico , Neoplasias da Glândula Tireoide/patologia , Tireotropina/genética , Tireotropina/metabolismo , Microambiente Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Biochim Biophys Acta Mol Cell Res ; 1866(7): 1124-1136, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30452936

RESUMO

Store operated Ca2+ entry (SOCE) is the most important Ca2+ entry pathway in non-excitable cells. However, SOCE can also play a pivotal role in excitable cells such as anterior pituitary (AP) cells. The AP gland contains five different cell types that release six major AP hormones controlling most of the entire endocrine system. AP hormone release is modulated by Ca2+ signals induced by different hypothalamic releasing hormones (HRHs) acting on specific receptors in AP cells. TRH and LHRH both induce Ca2+ release and Ca2+ entry in responsive cells while GHRH and CRH only induce Ca2+ entry. SOCE has been shown to contribute to Ca2+ responses induced by TRH and LHRH but no molecular evidence has been provided. Accordingly, we used AP cells isolated from mice devoid of Orai1 channels (noted as Orai1-/- or Orai1 KO mice) and mice lacking expression of all seven canonical TRP channels (TRPC) from TRPC1 to TRPC7 (noted as heptaTRPC KO mice) to investigate contribution of these putative channel proteins to SOCE and intracellular Ca2+ responses induced by HRHs. We found that thapsigargin-evoked SOCE is lost in AP cells from Orai1-/- mice but unaffected in cells from heptaTRPC KO mice. Conversely, while spontaneous intracellular Ca2+-oscillations related to electrical activity were not affected in the Orai1-/- mice, these responses were significantly reduced in heptaTRPC KO mice. We also found that Ca2+ entry induced by TRH and LHRH is decreased in AP cells isolated from Orai1-/-. In addition, Ca2+ responses to several HRHs, particularly TRH and GHRH, are decreased in the heptaTRPC KO mice. These results indicate that expression of Orai1, and not TRPC channel proteins, is necessary for thapsigargin-evoked SOCE and is required to support Ca2+ entry induced by TRH and LHRH in mouse AP cells. In contrast, TRPC channel proteins appear to contribute to spontaneous Ca2+-oscillations and Ca2+ responses induced by TRH and GHRH. We conclude that expression of Orai1 and TRPC channels proteins may play differential and significant roles in AP physiology and endocrine control.


Assuntos
Sinalização do Cálcio , Cálcio , Hormônio Liberador de Gonadotropina/metabolismo , Proteína ORAI1/deficiência , Adeno-Hipófise/metabolismo , Canais de Cátion TRPC/deficiência , Tireotropina/metabolismo , Animais , Hormônio Liberador de Gonadotropina/genética , Camundongos , Camundongos Knockout , Tireotropina/genética
7.
Cell Physiol Biochem ; 51(1): 142-153, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30448824

RESUMO

BACKGROUND/AIMS: Thyroid hormones have been recently linked to senescence and longevity. Given the recent description of TSHB mRNA in human adipose tissue (AT), we aimed to investigate the relationship between local AT TSH and adipose tissue senescence. METHODS: TSHB mRNA (measured by real-time PCR) and markers of adipose tissue senescence [BAX, DBC1, TP53, TNF (real-time PCR), telomere length (Telo TAGGG Telomere Length Assay) and lipidomics (liquid chromatography mass spectrometry)] were analysed in subcutaneous (SAT) and visceral (VAT) AT from euthyroid subjects. The chronic effects of TSH were also investigated in AT from hypothyroid rats and after recombinant human TSH (rhTSH) administration in human adipocytes. RESULTS: Both VAT and SAT TSHB gene expression negatively correlated with markers of AT cellular senescence (BAX, DBC1, TP53, TNF gene expression and specific glucosylceramides) and positively associated with telomere length. Supporting these observations, both rhTSH administration in human adipocytes and increased TSH in hypothyroid rats resulted in decreased markers of cellular senescence (Bax and Tp53 mRNA) in both gonadal and subcutaneous white adipose tissue. CONCLUSION: These data point to a possible role of TSH in AT cellular senescence.


Assuntos
Senescência Celular , Hipotireoidismo/patologia , Gordura Intra-Abdominal/metabolismo , Gordura Subcutânea/metabolismo , Tireotropina Subunidade beta/metabolismo , Adulto , Animais , Biomarcadores/metabolismo , Glicemia/análise , Senescência Celular/efeitos dos fármacos , Senescência Celular/genética , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hipotireoidismo/veterinária , Gordura Intra-Abdominal/citologia , Gordura Intra-Abdominal/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Ratos , Ratos Sprague-Dawley , Gordura Subcutânea/citologia , Gordura Subcutânea/efeitos dos fármacos , Homeostase do Telômero , Tireotropina/genética , Tireotropina/metabolismo , Tireotropina/farmacologia , Tireotropina Subunidade beta/genética , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
8.
Mutagenesis ; 33(5-6): 351-357, 2018 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-30481337

RESUMO

As apoptosis and genome instability in children with autoimmune diseases (AIDs) are insufficiently investigated, we aimed to analyse them in peripheral blood lymphocytes (PBLs) of children and adolescents with Hashimoto's thyroiditis (HT), Graves' disease (GD) and type 1 diabetes mellitus (T1DM), including possible factors that could affect their occurrence. The study population included 24 patients and 19 healthy controls. Apoptotic cells were detected using an Annexin V-FITC/7-AAD kit. Genome instability was measured as micronuclei (MNs) frequency using the cytokinesis-block MN assay. In addition, comet assay was performed for determination of genome instability as genome damage index (GDI) in new subpopulation of patients with T1DM. The percentage of apoptotic PBLs in patients with AID was significantly lower than in control subjects. There was a positive correlation between thyroid-stimulating homone (TSH) concentration and the proportion of cells in late stage apoptosis in patients with autoimmune thyroid diseases (AITDs). The MN frequency in patients was significantly higher than in controls. Individuals with HT or T1DM had a significantly higher MN frequency than those with GD. Similarly, the value of GDI in patients with T1DM was significantly higher than in controls. The level of apoptosis was positively correlated with MN frequency as well as with GDI in patients with AID. In conclusion, children with AITD (HT and GD) and T1DM have a significantly lower level of apoptosis in PBLs and significantly higher MN frequency as GDI than healthy subjects. Apoptosis and the level of genome instability in these patients with AID are positively correlated.


Assuntos
Apoptose/genética , Doenças Autoimunes/genética , Instabilidade Genômica/genética , Testes para Micronúcleos , Adolescente , Anexina A5/genética , Doenças Autoimunes/imunologia , Doenças Autoimunes/fisiopatologia , Criança , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/fisiopatologia , Feminino , Instabilidade Genômica/imunologia , Doença de Graves/genética , Doença de Graves/imunologia , Doença de Graves/fisiopatologia , Doença de Hashimoto/genética , Doença de Hashimoto/imunologia , Doença de Hashimoto/fisiopatologia , Humanos , Linfócitos/patologia , Tireotropina/genética
9.
J Comp Neurol ; 526(15): 2444-2461, 2018 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-30242838

RESUMO

We recently reported that the number of hypothalamic tanycytes expressing pro-opiomelanocortin (Pomc) is highly variable among brains of adult rats. While its cause and significance remain unknown, identifying other variably expressed genes in tanycytes may help understand this curious phenomenon. In this in situ hybridization study, we report that the Prss56 gene, which encodes a trypsin-like serine protease and is expressed in neural stem/progenitor cells, shows a similarly variable mRNA expression in tanycytes of adult rats and correlates inversely with tanycyte Pomc mRNA. Prss56 was expressed in α1, ß1, subsets of α2, and some median eminence γ tanycytes, but virtually absent from ß2 tanycytes. Prss56 was also expressed in vimentin positive tanycyte-like cells in the parenchyma of the ventromedial and arcuate nuclei, and in thyrotropin beta subunit-expressing cells of the pars tuberalis of the pituitary. In contrast to adults, Prss56 expression was uniformly high in tanycytes in adolescent rats. In mice, Prss56-expressing tanycytes and parenchymal cells were also observed but fewer in number and without significant variations. The results identify Prss56 as a second gene that is expressed variably in tanycytes of adult rats. We propose that the variable, inversely correlating expression of Prss56 and Pomc reflect periodically oscillating gene expression in tanycytes rather than stable expression levels that vary between individual rats. A possible functional link between Prss56 and POMC, and Prss56 as a potential marker for migrating tanycytes are discussed.


Assuntos
Células Ependimogliais/metabolismo , Hipotálamo/metabolismo , Pró-Opiomelanocortina/biossíntese , Pró-Opiomelanocortina/genética , Serina Proteases/biossíntese , Serina Proteases/genética , Envelhecimento/metabolismo , Animais , Contagem de Células , Células Ependimogliais/classificação , Feminino , Regulação da Expressão Gênica , Hipotálamo/química , Antígeno Ki-67/metabolismo , Masculino , Hipófise/metabolismo , Ratos , Ratos Sprague-Dawley , Serina Proteases/metabolismo , Terminologia como Assunto , Tireotropina/biossíntese , Tireotropina/genética
10.
Sci Rep ; 8(1): 10090, 2018 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-29973617

RESUMO

Deiodinase 2 (DIO2) plays an important role in thyroid hormone metabolism and its regulation. However, molecular mechanism that regulates DIO2 activity remains unclear; only mutaions in selenocysteine insertion sequence binding protein 2 and selenocysteine tranfer RNA (tRNA[Ser]Sec) are reported to result in decreased DIO2 activity. Two patients with clinical evidence of abnormal thyroid hormone metabolism were identified and found to have TSHR mutations as well as DIO2 T92A single nucleotide polymorphism (SNP). Primary-cultured fibroblasts from one patient present a high level of basal DIO2 enzymatic activity, possibly due to compensation by augmented DIO2 expression. However, this high enzymatic active state yet fails to respond to accelerating TSH. Consequently, TSHR mutations along with DIO2 T92A SNP ("double hit") may lead to a significant reduction in DIO2 activity stimulated by TSH, and thereby may have clinical relevance in a select population of hypothyroidism patients who might benefit from a T3/T4 combination therapy.


Assuntos
Hipotireoidismo/genética , Iodeto Peroxidase/genética , Receptores da Tireotropina/genética , Doenças da Glândula Tireoide/genética , Idoso , AMP Cíclico/genética , Feminino , Regulação da Expressão Gênica/genética , Humanos , Hipotireoidismo/metabolismo , Hipotireoidismo/patologia , Iodeto Peroxidase/metabolismo , Masculino , Mutação , Polimorfismo de Nucleotídeo Único/genética , Doenças da Glândula Tireoide/metabolismo , Doenças da Glândula Tireoide/patologia , Tireotropina/genética , Tireotropina/metabolismo , Tiroxina/genética , Tiroxina/metabolismo , Tri-Iodotironina/genética , Tri-Iodotironina/metabolismo
11.
Iran J Allergy Asthma Immunol ; 17(2): 158-170, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29757589

RESUMO

The production of human thyroid stimulating hormone (hTSH) immunoassays requires specific antibodies against hTSH which is a cumbersome process. Therefore, producing specific polyclonal antibodies against engineered recombinant fusion hTSH antigens would be of great significance. The best immunogenic region of the hTSH was selected based on in silico analyses and equipped with two different fusions. Standard methods were used for protein expression, purification, verification, structural evaluation, and immunizations of the white New Zealand rabbits. Ultimately, immunized serums were used for antibody titration, purification and characterization (specificity, sensitivity and cross reactivity). The desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. Structural analyses indicated that only the bigger antigen has showed changed 2 dimensional (2D) and 3D structural properties in comparison to the smaller antigen. The raised polyclonal antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum and negligible. The fusion which was solely composed of the tetanus toxin epitopes led to better protein folding and was capable of immunizing the host animals resulting into high titer antibody. Therefore, the minimal fusion sequences seem to be more effective in eliciting specific antibody responses.


Assuntos
Anticorpos/imunologia , Tireotropina/imunologia , Sequência de Aminoácidos , Animais , Especificidade de Anticorpos , Sequência de Bases , Clonagem Molecular , Reações Cruzadas , Epitopos , Feminino , Humanos , Imunização , Conformação Proteica , Coelhos , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo , Toxina Tetânica/química , Toxina Tetânica/genética , Toxina Tetânica/imunologia , Toxina Tetânica/metabolismo , Tireotropina/química , Tireotropina/genética , Tireotropina/metabolismo
12.
Best Pract Res Clin Endocrinol Metab ; 32(3): 241-256, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29779579

RESUMO

The thyroid gland produces thyroid hormones (TH), which are essential regulators for growth, development and metabolism. The thyroid is mainly controlled by the thyroid-stimulating hormone (TSH) that binds to its receptor (TSHR) on thyrocytes and mediates its action via different G protein-mediated signaling pathways. TSH primarily activates the Gs-pathway, and at higher concentrations also the Gq/11-pathway, leading to an increase of intracellular cAMP and Ca2+, respectively. To date, the physiological importance of other G protein-mediated signaling pathways in thyrocytes is unclear. Congenital hypothyroidism (CH) is defined as the lack of TH at birth. In familial cases, high-throughput sequencing methods have facilitated the identification of novel mutations. Nevertheless, the precise etiology of CH yet remains unraveled in a proportion of cases. Genetically modified mouse models can reveal new pathophysiological mechanisms of thyroid diseases. Here, we will present an overview of genetic mouse models for thyroid diseases, which have provided crucial insights into thyroid gland development, function, and growth with a special focus on TSHR and microRNA signaling.


Assuntos
Camundongos Transgênicos , Glândula Tireoide/embriologia , Glândula Tireoide/crescimento & desenvolvimento , Glândula Tireoide/fisiologia , Animais , Hipotireoidismo Congênito/genética , Hipotireoidismo Congênito/patologia , Humanos , Camundongos , Receptores da Tireotropina/genética , Receptores da Tireotropina/fisiologia , Transdução de Sinais/fisiologia , Doenças da Glândula Tireoide/genética , Doenças da Glândula Tireoide/patologia , Hormônios Tireóideos/fisiologia , Tireotropina/genética , Tireotropina/fisiologia
13.
Biol Trace Elem Res ; 186(1): 85-90, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29546542

RESUMO

Thyroid-stimulating hormone (TSH) is secreted by the pituitary gland and promotes thyroid growth and function, with increased TSH levels typically associated with hypothyroidism. By consulting the literature, we found that the TSHR, PAX8, and PDE4B genes are associated with thyroid function. Recently, copy number variations (CNVs) have been used as genetic markers to investigate inter-individual variation. Therefore, we investigated the relationship between the TSHR, PAX8, and PDE4B gene CNVs and TSH abnormalities, by calculating variations in gene copy number. Four hundred and eighty-one participants, 232 healthy controls and 249 patients with TSH abnormalities, were selected from three distinct areas in China with different iodine statuses. RT-PCR was used to detect CNVs. Urinary iodine concentrations (UIC) were measured by As3+-Ce4+ catalytic spectrophotometry. There was an association between a CNV at the TSHR gene and TSH abnormalities (p = 0.002). The distribution of PAX8 and PDE4B gene CNVs between patients with TSH abnormalities and healthy controls was not significantly different. UIC > 200 µg/l (OR = 1.49, 95% CI = 1.01-2.22) and the TSHR gene (OR = 6.01, 95% CI = 1.96-18.41) were found to be risk factors for TSH abnormalities. PAX8 and PDE4B gene CNVs were not significantly associated with TSH abnormalities. There was no significant interaction between UIC and any of the examined CNVs. In conclusion, the TSHR gene CNV was associated with the development of TSH abnormalities. No significant associations were revealed between urinary iodine levels and candidate gene CNVs.


Assuntos
Variações do Número de Cópias de DNA/genética , Receptores da Tireotropina/genética , Tireotropina/genética , China , Feminino , Humanos , Iodo/urina , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real
14.
Bull Exp Biol Med ; 164(4): 430-433, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29500802

RESUMO

Sex differences in the expression of iodide transporter SLC5A5 and thyroid peroxidase in thyroid follicular epithelium and thyroid serum profile were assessed in pubertal rats exposed to endocrine disruptor DDT starting from the first postnatal day. It was found that exposure to DDT reduced expression of SLC5A5 in peripheral regions of thyroid lobes in males and in central regions in females. The most pronounced sex differences were observed in thyroid peroxidase expression that remained sensitive to thyroid stimulating hormone regulation in males and lost sensitivity to pituitary stimulation in females after exposure to disruptor, which determines more pronounced hypothyroidism in females.


Assuntos
DDT/farmacologia , Disruptores Endócrinos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hipotireoidismo/genética , Glândula Tireoide/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Feminino , Hipotireoidismo/induzido quimicamente , Hipotireoidismo/metabolismo , Hipotireoidismo/patologia , Iodeto Peroxidase/genética , Iodeto Peroxidase/metabolismo , Masculino , Ratos , Ratos Wistar , Receptores da Tireotropina/genética , Receptores da Tireotropina/metabolismo , Fatores Sexuais , Maturidade Sexual , Simportadores/genética , Simportadores/metabolismo , Glândula Tireoide/metabolismo , Glândula Tireoide/patologia , Tireotropina/genética , Tireotropina/metabolismo , Tiroxina/genética , Tiroxina/metabolismo , Tri-Iodotironina/genética , Tri-Iodotironina/metabolismo
15.
Toxicol Lett ; 285: 81-86, 2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29305326

RESUMO

Bisphenol A (BPA) is a component of polycarbonate plastics, epoxy resins and polystyrene found in many common products. Several reports revealed potent in vivo and in vitro effects. In this study we analyzed the effects of the exposure to BPA in the hypothalamic-pituitary-thyroid axis in female rats, both in vivo and in vitro. Female Sprague-Dawley rats were injected sc from postnatal day 1 (PND1) to PND10 with BPA: 500 µg 50 µl-1 oil (B500), or 50 µg 50 µl-1 (B50), or 5 µg 50 µl-1 (B5). Controls were injected with 50 µl vehicle during the same period. Neonatal exposure to BPA did not modify TSH levels in PND13 females, but it increased them in adults in estrus. Serum T4 was lower in B5 and B500 with regards to Control, whereas no difference was seen in T3. No significant differences were observed in TRH, TSHß and TRH receptor expression between groups. TSH release from PPC obtained from adults in estrus was also higher in B50 with regard to Control. In vitro 24 h pre-treatment with BPA or E2 increased basal TSH as well as prolactin release. On the other hand, both BPA and E2 lowered the response to TRH. The results presented here show that the neonatal exposure to BPA alters the hypothalamic pituitary-thyroid axis in adult rats in estrus, possibly with effects on the pituitary and thyroid. They also show that BPA alters TSH release from rat PPC through direct actions on the pituitary.


Assuntos
Compostos Benzidrílicos/toxicidade , Disruptores Endócrinos/toxicidade , Hipotálamo/efeitos dos fármacos , Fenóis/toxicidade , Hipófise/efeitos dos fármacos , Glândula Tireoide/efeitos dos fármacos , Envelhecimento/sangue , Envelhecimento/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Hipófise/crescimento & desenvolvimento , Hipófise/metabolismo , Ratos Sprague-Dawley , Receptores do Hormônio Liberador da Tireotropina/genética , Receptores do Hormônio Liberador da Tireotropina/metabolismo , Glândula Tireoide/crescimento & desenvolvimento , Glândula Tireoide/metabolismo , Tireotropina/sangue , Tireotropina/genética , Hormônio Liberador de Tireotropina/sangue
16.
Horm Cancer ; 9(1): 1-11, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29209896

RESUMO

Oncogene-induced senescence (OIS) explains the phenomenon of cellular senescence triggered by the action of oncogenes. It is a mechanism adopted by a cell to inhibit progression of benign tumors into malignancy, occurs in premalignant lesions, and is almost never present in malignant lesions. BRAF mutations occur in about 40-45% of all papillary thyroid carcinomas (PTCs) and of which 99.7% is the BRAFV600E mutation. A unique phenotype of the BRAFV600E mutation is the upregulation of the thyroid-stimulating hormone receptor (TSHR) on thyrocyte membranes. Despite the overexpression of the receptor, BRAFV600E cells undergo cell cycle arrest leading to OIS via a negative feedback signaling mechanism. A simultaneous increase in serum thyroid-stimulating hormone (TSH) in response to hypothyroidism (common in autoimmune diseases such as Hashimoto's thyroiditis) would cause senescent tumor cells to overcome OIS and proceed towards malignancy, hence showing the importance of TSH/TSHR signaling in the development of PTCs. Increase in TSH/TSHR signaling triggers an increase in levels of downstream enzymes such as manganese superoxide dismutase (MnSOD) and dual-specific phosphatase 6 (DUSP6) which eventually results in the production of oncogenic proteins such as c-Myc. Therefore, the detection of these genetic alterations as effective biomarkers for premalignant lesions of PTC is important in clinical settings and techniques such as polymerase chain reaction-mediated restriction fragment length polymorphism (PCR-RFLP) and real-time PCR can be used to detect the BRAFV600E point mutation and overexpression of TSHR, MnSOD, and DUSP6, respectively.


Assuntos
Senescência Celular/genética , Fosfatase 6 de Especificidade Dupla/genética , Proteínas Proto-Oncogênicas B-raf/genética , Câncer Papilífero da Tireoide/genética , Tireotropina/genética , Progressão da Doença , Humanos , Mutação , Proteínas Proto-Oncogênicas c-myc/genética , Receptores Acoplados a Proteínas-G/genética , Transdução de Sinais/genética , Superóxido Dismutase/genética , Câncer Papilífero da Tireoide/patologia
17.
Usp Fiziol Nauk ; 48(1): 80-90, 2017.
Artigo em Russo | MEDLINE | ID: mdl-29283520

RESUMO

The review provides information about the features of the sensitivity of thymocytes, lymphoid organs' cells and T-lymphocytes of peripheral blood to the hormones secreted by anterior pituitary gland's cells: growth hormone, thyrotropin, adrenocorticotropic hormone, prolactin and ß-endorphin. Some aspects of the T-lymphocytes's response to humoral signals from the hypophysis are shown in the article. Also the pituitary hormones' role in the regulation of proliferation, differentiation, and cytokine production of T-lymphocytes in normal and pathological conditions of the organism being discussed.


Assuntos
Hormônio Adrenocorticotrópico/farmacologia , Hormônio do Crescimento/farmacologia , Adeno-Hipófise/metabolismo , Prolactina/farmacologia , Timócitos/efeitos dos fármacos , Tireotropina/farmacologia , beta-Endorfina/farmacologia , Hormônio Adrenocorticotrópico/genética , Hormônio Adrenocorticotrópico/imunologia , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação da Expressão Gênica , Hormônio do Crescimento/genética , Hormônio do Crescimento/imunologia , Humanos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Cultura Primária de Células , Prolactina/genética , Prolactina/imunologia , Transdução de Sinais , Timócitos/citologia , Timócitos/imunologia , Tireotropina/genética , Tireotropina/imunologia , beta-Endorfina/genética , beta-Endorfina/imunologia
18.
J Cell Sci ; 130(24): 4155-4167, 2017 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-29093023

RESUMO

Many secretory cells increase the synthesis and secretion of cargo proteins in response to specific stimuli. How cells couple increased cargo load with a coordinate rise in secretory capacity to ensure efficient transport is not well understood. We used thyroid cells stimulated with thyrotropin (TSH) to demonstrate a coordinate increase in the production of thyroid-specific cargo proteins and ER-Golgi transport factors, and a parallel expansion of the Golgi complex. TSH also increased expression of the CREB3L1 transcription factor, which alone caused amplified transport factor levels and Golgi enlargement. Furthermore, CREB3L1 potentiated the TSH-induced increase in Golgi volume. A dominant-negative CREB3L1 construct hampered the ability of TSH to induce Golgi expansion, implying that this transcription factor contributes to Golgi expansion. Our findings support a model in which CREB3L1 acts as a downstream effector of TSH to regulate the expression of cargo proteins, and simultaneously increases the synthesis of transport factors and the expansion of the Golgi to synchronize the rise in cargo load with the amplified capacity of the secretory pathway.


Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Complexo de Golgi/genética , Proteínas do Tecido Nervoso/genética , Glândula Tireoide/metabolismo , Tireotropina/genética , Linhagem Celular , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Retículo Endoplasmático/genética , Retículo Endoplasmático/metabolismo , Regulação da Expressão Gênica/genética , Complexo de Golgi/metabolismo , Humanos , Proteínas do Tecido Nervoso/metabolismo , Via Secretória/genética , Tireotropina/metabolismo
19.
J Clin Invest ; 127(12): 4326-4337, 2017 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-29083325

RESUMO

Deficiency in Krüppel-like zinc finger transcription factor GLI-similar 3 (GLIS3) in humans is associated with the development of congenital hypothyroidism. However, the functions of GLIS3 in the thyroid gland and the mechanism by which GLIS3 dysfunction causes hypothyroidism are unknown. In the current study, we demonstrate that GLIS3 acts downstream of thyroid-stimulating hormone (TSH) and TSH receptor (TSHR) and is indispensable for TSH/TSHR-mediated proliferation of thyroid follicular cells and biosynthesis of thyroid hormone. Using ChIP-Seq and promoter analysis, we demonstrate that GLIS3 is critical for the transcriptional activation of several genes required for thyroid hormone biosynthesis, including the iodide transporters Nis and Pds, both of which showed enhanced GLIS3 binding at their promoters. The repression of cell proliferation of GLIS3-deficient thyroid follicular cells was due to the inhibition of TSH-mediated activation of the mTOR complex 1/ribosomal protein S6 (mTORC1/RPS6) pathway as well as the reduced expression of several cell division-related genes regulated directly by GLIS3. Consequently, GLIS3 deficiency in a murine model prevented the development of goiter as well as the induction of inflammatory and fibrotic genes during chronic elevation of circulating TSH. Our study identifies GLIS3 as a key regulator of TSH/TSHR-mediated thyroid hormone biosynthesis and proliferation of thyroid follicular cells and uncovers a mechanism by which GLIS3 deficiency causes neonatal hypothyroidism and prevents goiter development.


Assuntos
Proliferação de Células , Receptores da Tireotropina/metabolismo , Proteínas Repressoras/metabolismo , Glândula Tireoide/metabolismo , Hormônios Tireóideos/biossíntese , Tireotropina/metabolismo , Transativadores/metabolismo , Animais , Bócio/genética , Bócio/metabolismo , Bócio/prevenção & controle , Alvo Mecanístico do Complexo 1 de Rapamicina/genética , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Camundongos , Camundongos Knockout , Regiões Promotoras Genéticas , Receptores da Tireotropina/genética , Proteínas Repressoras/genética , Proteínas Ribossômicas/genética , Proteínas Ribossômicas/metabolismo , Transportadores de Sulfato/genética , Transportadores de Sulfato/metabolismo , Simportadores/genética , Simportadores/metabolismo , Glândula Tireoide/citologia , Hormônios Tireóideos/genética , Tireotropina/genética , Transativadores/genética
20.
Cell Mol Biol (Noisy-le-grand) ; 63(8): 93-94, 2017 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-28886330

RESUMO

Cornelia de Lange syndrome (CdLs), which is also called Brachmann de Lange syndrome, is a congenital disorder characterized by distinctive facial features, prenatal and postnatal growth deficiency, feeding difficulties, psychomotor delay, behavioral problems, and associated malformations that mainly involve the upper extremities. The prevalence ranges from 1:100,000 to as high as 1:10,000. Most cases (50-60%) were carried mutation in NIPBL gene. To our knowledge this is the first CdLs Indonesian case that reported with molecular analysis study. We present an 11 months old female Indonesian patient with classic CdLs with congenital hypothyroid. Genetics studies were performed in intron 1, exon 2, exon 10 and exon 22 of NIPBL gene. Thyroid studies (T3, T4, TSH and thyroid scan) were performed. Low level of T3 and T4, and high level of TSH were observed. Thyroid agenesis was found in thyroid scan examination. We detected thyroid agenesis which has been never reported in CdLs patients. We could not find any mutation in intron 1, exon 2, exon 10 and exon 22 of NIPBL gene. Further genetics examinations were necessary whether there is mutation in other locus.


Assuntos
Hipotireoidismo Congênito/diagnóstico , Síndrome de Lange/diagnóstico , Disgenesia da Tireoide/diagnóstico , Proteínas de Ciclo Celular , Hipotireoidismo Congênito/genética , Hipotireoidismo Congênito/patologia , Síndrome de Lange/genética , Síndrome de Lange/patologia , Feminino , Expressão Gênica , Humanos , Indonésia , Lactente , Proteínas/genética , Proteínas/metabolismo , Disgenesia da Tireoide/genética , Disgenesia da Tireoide/patologia , Tireotropina/genética , Tireotropina/metabolismo , Tiroxina/deficiência , Tiroxina/genética , Tri-Iodotironina/deficiência , Tri-Iodotironina/genética , Regulação para Cima
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