RESUMEN
Di-(2-ethylhexyl) phthalate (DEHP) has the potential to disrupt the thyroid endocrine system, but the underlying mechanism is unknown. In this study, zebrafish (Danio rerio) embryos were exposed to different concentrations of DEHP (0, 40, 100, 200, 400 µg/L) from 2 to 168 hours post fertilization (hpf). Thyroid hormones (THs) levels and transcriptional profiling of key genes related to hypothalamus-pituitary-thyroid (HPT) axis were examined. The result of whole-body thyroxine (T4) and triiodothyronine (T3) indicated that the thyroid hormone homeostasis was disrupted by DEHP in the zebrafish larvae. After exposure to DEHP, the mRNA expressions of thyroid stimulating hormone (tshß) and corticotrophin releasing hormone (crh) genes were increased in a concentration dependent manner, respectively. The expression level of genes involved in thyroid development (nkx2.1 and pax8) and thyroid synthesis (sodium/iodide symporter, nis, thyroglobulin, tg) were also measured. The transcripts of nkx2.1 and tg were significantly increased after DEHP exposure, while those of nis and pax8 had no significant change. Down-regulation of uridinediphosphate-glucuronosyl-transferase (ugt1ab) and up-regulation of thyronine deiodinase (dio2) might change the THs levels. In addition, the transcript of transthyretin (ttr) was up-regulated, while the mRNA levels of thyroid hormone receptors (trα and trß) remained unchanged. All the results demonstrated that exposure to DEHP altered the whole-body thyroid hormones in the zebrafish larvae and changed the expression profiling of key genes related to HPT axis, proving that DEHP induced the thyroid endocrine toxicity and potentially affected the synthesis, regulation and action of thyroid hormones.
Asunto(s)
Dietilhexil Ftalato/toxicidad , Hipotálamo/embriología , Hipófisis/embriología , Glándula Tiroides/embriología , Pez Cebra/embriología , Animales , Hormona Liberadora de Corticotropina/sangre , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Proteínas Nucleares/biosíntesis , Factor de Transcripción PAX8/biosíntesis , Simportadores/biosíntesis , Tiroglobulina/biosíntesis , Factor Nuclear Tiroideo 1 , Tirotropina/sangre , Tiroxina/sangre , Factores de Transcripción/biosíntesis , Triyodotironina/sangre , Proteínas de Pez Cebra/biosíntesisRESUMEN
Graves' disease, the most common cause of hyperthyroidism, is an autoimmune disorder. Antithyroid drugs have been selected as the first-line treatment of Graves' disease in Korea, Japan, and European countries. However, antithyroid drugs such as methimazole (MMI) and prophylthiouracil (PTU) have limitations in clinical applications because of their side effects. In this study, we performed a clinical trial and in vitro study to investigate the clinical effects and action mechanism of Ahnjeonbaekho-tang (AJBHT), an herbal remedy for Graves' disease. In a clinical study of Graves' disease patients who had side effects from antithyroid drugs, we found that treatment by AJBHT resulted in a reduction of serum triiodothyronine (T3) and free thyroxine (FT4) levels and an increase in thyroid stimulating hormone (TSH) levels (T3: p<0.0001, FT4: p=0.0012, TSH: p=0.0370, respectively). In vitro, AJBHT significantly inhibits FRTL-5 cell proliferation, DNA synthesis, cyclic AMP production, T4 synthesis, and the expression of thyroglobulin (Tg) mRNA in comparison with the control. These results suggest that AJBHT might suppress T(4) synthesis by modulating adenosine 3',5'-cyclic monophosphate (cAMP) and Tg expression, and therefore, AJBHT could be an alternative therapy for Graves' disease patients who have side effects from antithyroid drugs.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Enfermedad de Graves/tratamiento farmacológico , Fitoterapia , Animales , Línea Celular , Proliferación Celular/efectos de los fármacos , AMP Cíclico/biosíntesis , ADN/biosíntesis , Enfermedad de Graves/metabolismo , Humanos , Yoduro Peroxidasa/biosíntesis , Yoduro Peroxidasa/sangre , Extractos Vegetales/farmacología , ARN/biosíntesis , ARN/genética , Ratas , Ratas Endogámicas F344 , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sales de Tetrazolio , Tiazoles , Tiroglobulina/biosíntesis , Tiroglobulina/sangre , Tirotropina/biosíntesis , Tirotropina/sangre , Tiroxina/biosíntesis , Tiroxina/sangreRESUMEN
We investigated whether recombinant human TSH (rhTSH) safely and effectively induces uptake of high-dose 131-iodine (131I) to ablate thyroid remnant or treat disease, in patients with well-differentiated thyroid carcinoma. Eleven consecutive patients unable to tolerate thyroid hormone withdrawal received one im injection of 0.9 mg rhTSH on 2 consecutive days before receiving 4000 MBq (approximately 108 mCi) radioiodine orally. Eight patients received one, and 3 patients 2 courses. Our series comprised 7 women and 4 men (mean age, 78 yr, range: 56-87 yr). Ten patients had undergone total or near-total thyroidectomy up to 19 yr earlier. rhTSH-stimulated single course radioiodine with the intention to ablate thyroid remnant was performed in 3 patients, with following estimation of radioiodine uptake and TG measurements. Of another 8 patients given this treatment palliatively, 5 had radiological, clinical and/or laboratory response, including: 80% decreased pathological uptake between treatment courses; pronounced decrease in bone pain; diminished symptoms; improved physical condition and quality of life; lower serum TG concentration; and/or normalization of TG recovery test. Two patients with small lung metastases on computed tomography had no detectable radioiodine uptake or other response; they also lacked uptake after withdrawal-stimulated radioiodine treatment. Despite being elderly and frail, patients generally tolerated treatment well; rhTSH caused nausea in one patient and transiently increased pain in bone and soft tissue lesions in another. We conclude that rhTSH-stimulated high-dose radioiodine for remnant ablation or tumor treatment is safe, feasible and seemingly effective, enhancing quality of life and offering reasonable palliation in patients with advanced disease.
Asunto(s)
Radioisótopos de Yodo/uso terapéutico , Neoplasias de la Tiroides/tratamiento farmacológico , Neoplasias de la Tiroides/radioterapia , Tirotropina/uso terapéutico , Anciano , Anciano de 80 o más Años , Terapia Combinada , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Radioisótopos de Yodo/administración & dosificación , Radioisótopos de Yodo/efectos adversos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/secundario , Masculino , Persona de Mediana Edad , Cuidados Paliativos , Proteínas Recombinantes/uso terapéutico , Seguridad , Tiroglobulina/biosíntesis , Neoplasias de la Tiroides/metabolismo , Tirotropina/efectos adversos , Insuficiencia del Tratamiento , Resultado del TratamientoRESUMEN
cDNA and genomic clones encoding human sarco/endoplasmic reticulum Ca(2+)-ATPase 3 (SERCA3) were isolated. The composite nucleotide sequence of the 4.6 kb cDNA, as well as the partial structure of 25 kb of genomic DNA encoding all but the 5' region of the gene, was determined. The nucleotide sequence coding for the last six amino acids of the pump and the 3'-untranslated region were identified within the sequence of the last exon. Northern blot hybridization analysis using cDNA probes derived from this exon detected a 4.8 kb transcript in several human tissues. Using a cDNA probe derived from the 5'-coding region an unexpected mRNA distribution pattern, consisting of two mRNA species of 4.8 and 4.0 kb, was detected in thyroid gland and bone marrow only. This is the first indication of an alternative splicing mechanism operating on the SERCA3 gene transcript, which most likely generates SERCA3 isoforms with altered C-termini. Human SERCA3 expressed in platelets and in COS cells transfected with the corresponding cDNA was detected with the previously described antibody N89 (directed against the N-terminal region of rat SERCA3) and with a new SERCA3-specific antiserum C91, directed against the extreme C-terminus of the human isoform. A monoclonal antibody PL/IM430, previously assumed to recognize SERCA3 in human platelets, does not react with the 97 kDa human SERCA3 transiently expressed in COS cells. Therefore the 97 kDa isoform detected by PL/IM430 more likely represents a novel SERCA pump, as recently suggested [Kovács, Corvazier, Papp, Magnier, Bredoux, Enyedi, Sarkadi and Enouf (1994) J. Biol. Chem. 269, 6177-6184]. Finally, by fluorescence in situ hybridization and chromosome G-banding analyses, the SERCA3 gene was assigned to human chromosome 17p13.3.
Asunto(s)
ATPasas Transportadoras de Calcio/biosíntesis , ATPasas Transportadoras de Calcio/genética , Cromosomas Humanos Par 13 , Retículo Endoplásmico/enzimología , Retículo Sarcoplasmático/enzimología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Línea Celular , Mapeo Cromosómico , Clonación Molecular , Sondas de ADN , ADN Complementario , Humanos , Hibridación Fluorescente in Situ , Cariotipificación , Datos de Secuencia Molecular , Especificidad de Órganos , Ratas , Mapeo Restrictivo , Homología de Secuencia de Aminoácido , Tiroglobulina/biosíntesis , Transcripción GenéticaRESUMEN
In the last two decades hyperthermia has increasingly been used as adjuvant therapy for the treatment of malignant tumours. The effects of heat were therefore analysed on cultured thyroid epithelial cells from patients with thyroid cancer and from non-malignant control thyroids. Purified thyroid cells were subjected to heat treatment (42.5 degrees C; 90 min). After 24 h [3H]thymidine incorporation was assessed and the expression of heat shock protein 72 (hsp72), thyroglobulin, CD54 (ICAM-I) and MHC class-Il were analysed by immunofluorescence staining. Additionally mRNA analysis was performed by Northern blotting. Whereas hyperthermia inhibited the proliferation of thyroid cells, it significantly increased the expression of hsp72, thyroglobulin, CD54 and HLA-DR (P < 0.05). Our results suggest that hyperthermia may suppress growth while supporting differentiation and immune recognition in thyroid cancer. It may therefore be beneficial as a treatment for patients with thyroid carcinoma.
Asunto(s)
Adenocarcinoma Folicular/patología , Carcinoma Papilar/patología , Carcinoma/patología , Hipertermia Inducida , Neoplasias de la Tiroides/patología , Adenocarcinoma Folicular/terapia , Adulto , Anciano , Northern Blotting , Western Blotting , Carcinoma/terapia , Carcinoma Papilar/terapia , División Celular , Línea Celular Transformada , Femenino , Técnica del Anticuerpo Fluorescente , Regulación Neoplásica de la Expresión Génica , Antígenos HLA-DR/biosíntesis , Proteínas HSP70 de Choque Térmico/biosíntesis , Proteínas HSP70 de Choque Térmico/genética , Calor , Humanos , Molécula 1 de Adhesión Intercelular/biosíntesis , Molécula 1 de Adhesión Intercelular/genética , Masculino , Persona de Mediana Edad , ARN Mensajero/análisis , Tiroglobulina/biosíntesis , Neoplasias de la Tiroides/terapia , Células Tumorales CultivadasRESUMEN
The synthesis is described of an N-terminal thyroglobulin (Tg) polypeptide of 27 kDa, which is capable of hormonogenesis, in a baculovirus system. This polypeptide was made using a 657 bp Tg cDNA cloned from human thyroid RNA by a polymerase chain reaction method. The cDNA contained the information for the Tg signal peptide, the N-terminally located site for thyroid hormone formation and, at the 3' end, a sequence coding for six histidine residues. The fragments produced were purified using a nickel-nitrilotriacetic acid column using these six histidine residues. The products were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and showed two glycosylated fragments of 32 and 34 kDa, both of which started with asparagine. Iodination of the fragments with lactoperoxidase in vitro resulted in the formation of thyroxine (T4). The formation rate of T4 in the fragments was about five times lower than that of the mature Tg dimer of 660 kDa, but ten times more rapid than in bovine serum albumin under the same conditions.
Asunto(s)
Nucleopoliedrovirus/genética , Proteínas Recombinantes de Fusión/biosíntesis , Tiroglobulina/biosíntesis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , ADN Complementario/genética , Regulación Viral de la Expresión Génica , Yodo/metabolismo , Lactoperoxidasa/metabolismo , Datos de Secuencia Molecular , Mariposas Nocturnas , Reacción en Cadena de la Polimerasa , Procesamiento Proteico-Postraduccional , Proteínas Recombinantes de Fusión/genética , Tiroglobulina/genética , Tiroxina/biosíntesisAsunto(s)
Masculino , Femenino , Humanos , Embarazo , Recién Nacido , Niño , Adulto , Hormonas/fisiología , Andrógenos/biosíntesis , Andrógenos/fisiología , Bombesina/biosíntesis , Calcitonina/biosíntesis , Calcitonina/fisiología , Calcitriol/biosíntesis , Calcitriol/fisiología , Ciclo Menstrual , Colecistoquinina/biosíntesis , Colecistoquinina/fisiología , Trastornos de la Menstruación/clasificación , Trastornos de la Menstruación/diagnóstico , Trastornos del Metabolismo del Calcio/diagnóstico , Trastornos del Metabolismo del Calcio/etiología , Trastornos del Metabolismo del Fósforo/diagnóstico , Trastornos del Metabolismo del Fósforo/etiología , Encefalinas/biosíntesis , Encefalinas/fisiología , Endorfinas/biosíntesis , Endorfinas/fisiología , Estrógenos/biosíntesis , Estrógenos/fisiología , Gastrinas/biosíntesis , Gastrinas/fisiología , Glucagón/antagonistas & inhibidores , Glucagón/biosíntesis , Glucagón/fisiología , Glucagonoma/diagnóstico , Glucagonoma/etiología , Glucocorticoides/biosíntesis , Glándula Tiroides , Glándula Tiroides/anatomía & histología , Glándulas Paratiroides , Glándulas Suprarrenales , Glándulas Suprarrenales/fisiología , Gonadotropinas/biosíntesis , Gonadotropinas/fisiología , Embarazo/fisiología , Hipotálamo , Hipotálamo/anatomía & histología , Hipófisis , Hipófisis/anatomía & histología , Hormona Adrenocorticotrópica/biosíntesis , Hormona Adrenocorticotrópica/fisiología , Hormona Paratiroidea , Hormona del Crecimiento/biosíntesis , Hormona del Crecimiento/fisiología , Hormonas Gastrointestinales/biosíntesis , Hormonas Gastrointestinales/fisiología , Hormonas Hipotalámicas/biosíntesis , Hormonas Hipotalámicas/fisiología , Hormonas Inhibidoras de la Liberación de Hormona Hipofisaria/fisiología , Hormonas Liberadoras de Hormona Hipofisaria/fisiología , Insulina/biosíntesis , Insulina/fisiología , Insulinoma/diagnóstico , Insulinoma/etiología , Yodo/deficiencia , Yodo/fisiología , Yodo/metabolismo , Lactógeno Placentario/biosíntesis , Lactógeno Placentario/fisiología , Menopausia/fisiología , Menstruación , Motilina/biosíntesis , Motilina/fisiología , Oxitocina/biosíntesis , Oxitocina/fisiología , Ovario , Ovario/anatomía & histología , Ovario/fisiología , Péptido Intestinal Vasoactivo/biosíntesis , Péptido Intestinal Vasoactivo/fisiología , Péptidos Similares al Glucagón/biosíntesis , Péptidos Similares al Glucagón/fisiología , Polipéptido Pancreático/biosíntesis , Polipéptido Pancreático/fisiología , Progesterona/biosíntesis , Progesterona/fisiología , Prolactina/biosíntesis , Prolactina/fisiología , Páncreas/anatomía & histología , Páncreas/embriología , Relaxina/biosíntesis , Relaxina/fisiología , Secretina/biosíntesis , Secretina/fisiología , Somatostatina/biosíntesis , Somatostatina/fisiología , Testosterona/biosíntesis , Testosterona/fisiología , Testículo/anatomía & histología , Testículo/citología , Testículo/fisiología , Tiroglobulina/biosíntesis , Tiroglobulina/fisiología , Tiroglobulina/metabolismo , Valores de Referencia , Vasopresinas/biosíntesis , Vasopresinas/fisiología , beta-Lipotropina/biosíntesisAsunto(s)
Humanos , Masculino , Femenino , Embarazo , Recién Nacido , Niño , Adulto , Hormonas/fisiología , Hormonas Hipotalámicas/biosíntesis , Hormonas Hipotalámicas/fisiología , Hipotálamo/anatomía & histología , Hipotálamo , Hormonas Liberadoras de Hormona Hipofisaria/fisiología , Hormonas Inhibidoras de la Liberación de Hormona Hipofisaria/fisiología , Hormona del Crecimiento/biosíntesis , Hormona del Crecimiento/fisiología , Prolactina/biosíntesis , Prolactina/fisiología , Lactógeno Placentario/biosíntesis , Lactógeno Placentario/fisiología , Hormona Adrenocorticotrópica/biosíntesis , Hormona Adrenocorticotrópica/fisiología , beta-Lipotropina/biosíntesis , Endorfinas/biosíntesis , Endorfinas/fisiología , Vasopresinas/biosíntesis , Vasopresinas/fisiología , Oxitocina/biosíntesis , Oxitocina/fisiología , Hipófisis/anatomía & histología , Hipófisis , Glándula Tiroides/anatomía & histología , Glándula Tiroides , Tiroglobulina/biosíntesis , Tiroglobulina/fisiología , Tiroglobulina/metabolismo , Yodo/deficiencia , Yodo/fisiología , Yodo/metabolismo , Glándulas Suprarrenales/fisiología , Glándulas Suprarrenales , Glucocorticoides/biosíntesis , Glándulas Paratiroides , Hormona Paratiroidea , Calcitonina/biosíntesis , Calcitonina/fisiología , Calcitriol/biosíntesis , Calcitriol/fisiología , Trastornos del Metabolismo del Calcio/diagnóstico , Trastornos del Metabolismo del Calcio/etiología , Trastornos del Metabolismo del Fósforo/diagnóstico , Trastornos del Metabolismo del Fósforo/etiología , Ovario/anatomía & histología , Ovario/fisiología , Ovario , Estrógenos/biosíntesis , Estrógenos/fisiología , Progesterona/biosíntesis , Progesterona/fisiología , Relaxina/biosíntesis , Relaxina/fisiología , Gonadotropinas/biosíntesis , Gonadotropinas/fisiología , Ciclo Menstrual , Menstruación , Trastornos de la Menstruación/clasificación , Trastornos de la Menstruación/diagnóstico , Menopausia/fisiología , Embarazo/fisiología , Testículo/anatomía & histología , Testículo/citología , Testículo/fisiología , Andrógenos/biosíntesis , Andrógenos/fisiología , Testosterona/biosíntesis , Testosterona/fisiología , Páncreas/anatomía & histología , Páncreas/embriología , Glucagón/antagonistas & inhibidores , Glucagón/biosíntesis , Glucagón/fisiología , Insulina/biosíntesis , Insulina/fisiología , Polipéptido Pancreático/biosíntesis , Polipéptido Pancreático/fisiología , Insulinoma/diagnóstico , Insulinoma/etiología , Glucagonoma/diagnóstico , Glucagonoma/etiología , Somatostatina/biosíntesis , Somatostatina/fisiología , Hormonas Gastrointestinales/biosíntesis , Hormonas Gastrointestinales/fisiología , Secretina/biosíntesis , Secretina/fisiología , Colecistoquinina/biosíntesis , Colecistoquinina/fisiología , Gastrinas/biosíntesis , Gastrinas/fisiología , Péptidos Similares al Glucagón/biosíntesis , Péptidos Similares al Glucagón/fisiología , Encefalinas/biosíntesis , Encefalinas/fisiología , Péptido Intestinal Vasoactivo/biosíntesis , Péptido Intestinal Vasoactivo/fisiología , Motilina/biosíntesis , Motilina/fisiología , Bombesina/biosíntesis , Valores de ReferenciaRESUMEN
In vitro studies using thyroid slices from human non-toxic goitres and from thyrotoxic glands show retarded incorporation of [14C]mannose into the 19S protein of thyrotoxic glands. This was not found using [14C]galactose with thyrotoxic glands or using either labelled sugar with slices from non-toxic goitres. Experiments with thyroid tissue from rats on a variety of treatment regimes such as iodine supplements, carbimazole alone or with iodine supplements did not show this differential delay of [14C]mannose incorporation. This suggests that there may be some abnormality of carbohydrate incorporation into thyroglobulin in thyrotoxicosis.
Asunto(s)
Hipertiroidismo/metabolismo , Manosa/metabolismo , Tiroglobulina/biosíntesis , Animales , Carbimazol/farmacología , Galactosa/metabolismo , Bocio/metabolismo , Humanos , Técnicas In Vitro , Yodo/metabolismo , Ratas , Glándula Tiroides/metabolismoRESUMEN
Lithium and excess iodide inhibit the release of thyroid hormone from preformed stores. We thus tested the hypothesis that this was due to an inhibition of thyroglobulin breakdown. Rats were pre-treated with propyl-thiouracil (PTU) for 3 weeks in order to deplete their thyroids of thyroglobulin. While the PTU was continued, lithium chloride (0.25 mEq./100 g weight) or potassium iodide (3 mg per rat) were injected every 12 h for d days. Thereafter the thyroglobulin content in thyroid gland homogenates was measured. PTU pre-treatment lowered the thyroglobulin content from 4.21 to 0.22 mg/100 mg gland. Lithium caused a marked re-accumulation of thyroglobulin to 0.60 mg/100 mg within 3 days. While iodide alone had only a borderline effect, it markedly potentiated the action of lithium and a combination of the two drugs increased the thyroglobulin content to 1.04 mg/100 mg. Thyroxine was injected into similarly pre-treated animals to suppress secretion of thyrotrophic hormone. This markedly inhibited the proteolysis of thyroglobulin and 1.3 mg/100 mg gland accumulated after 3 days. Excess iodide, given in addition to thyroxine, decreased the amount of thyroglobulin accumulated to 0.75 mg/100 mg gland. To study whether this could be explained by an inhibitory action of iodide on thyroglobulin biosynthesis, thyroid glands from animals treated with excess iodide were incubated in vitro in the presence of 0.2 mM iodide for 3 h. Iodide decreased the incorporation of radioactive leucine into total thyroidal protein and into thyroglobulin by 25 and 35% respectively. Iodide did not inhibit protein synthesis in the kidney, liver or muscle tissue. Thus, large doses of iodide selectively inhibit thyroglobulin biosynthesis.