RESUMO
BACKGROUND: The secretory activity of Sertoli cells (SC) is dependent on ion channel functions and protein synthesis and is critical to ongoing spermatogenesis. The aim of this study was to investigate the mechanism of action associated with a non-metabolizable amino acid [14C]-MeAIB (alpha-(methyl-amino)isobutyric acid) accumulation stimulated by T4 and the role of the integrin receptor in this event, and also to clarify whether the T4 effect on MeAIB accumulation and on Ca2+ influx culminates in cell secretion. METHODS: We have studied the rapid and plasma membrane initiated effects of T4 by using 45Ca2+ uptake and [45C]-MeAIB accumulation assays, respectively. Thymidine incorporation into DNA was used to monitor nuclear activity and quinacrine to analyze the secretory activity on SC. RESULTS: The stimulation of MeAIB accumulation byT4 appears to be mediated by the integrin receptor in the plasma membrane since tetrac and RGD peptide were able to nullify the effect of this hormone. In addition, T4 increases extracellular Ca2+ uptake and Ca2+ from intracellular stocks to enhance nuclear activity, but this genomic action seems not to influence SC secretion mediated by T4. Also, the cytoskeleton and CIC-3 chloride channel contribute to the membrane-associated responses of SC. CONCLUSIONS: T4 integrin receptor activation ultimately determines the plasma membrane responses on amino acid transport in SC, but it is not involved in calcium influx, cell secretion or the nuclear effect of the hormone. GENERAL SIGNIFICANCE: The integrin receptor activation by T4 may take a role in plasma membrane processes involved in the male reproductive system.
Assuntos
Membrana Celular/efeitos dos fármacos , Integrina alfaVbeta3/genética , Receptores de Superfície Celular/genética , Células de Sertoli/efeitos dos fármacos , Espermatogênese/fisiologia , Tiroxina/farmacologia , Aminoácidos/metabolismo , Ácidos Aminoisobutíricos/metabolismo , Animais , Animais Recém-Nascidos , Transporte Biológico/efeitos dos fármacos , Cálcio/metabolismo , Radioisótopos de Carbono , Canais de Cloreto/metabolismo , Citoesqueleto/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Integrina alfaVbeta3/metabolismo , Masculino , Oligopeptídeos/farmacologia , Ratos , Ratos Wistar , Receptores de Superfície Celular/metabolismo , Células de Sertoli/citologia , Células de Sertoli/fisiologia , Transdução de Sinais/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Timidina/metabolismo , Tiroxina/análogos & derivadosRESUMO
Inhibition or reversion of protein self-aggregation has been suggested as a possible preventive mechanism against amyloid diseases, and many efforts are underway to found out molecules capable to restrain the protein aggregation process. In this paper, the inhibitory effects of thyroid hormone analogues on heat-induced fibrillation process of serum albumin are reported. Among the analogues tested, 3,5,3',5'-tetraiodothyroacetic and 3,5,3'-triiodothyroacetic acid showed the most important inhibitory effects on amyloid formation. Thyroxine exhibits a lesser protective effect, while 3,5,3'-triiodothyronine showed no significant inhibition. The gaining of a negative charge together with a size reduction of the hormone molecule could play an essential role in the inhibition of fibrils formation. According to infrared spectroscopy results, the thyroid hormones analogues protective effects proceed via the stabilization of the protein native structure. The current work demonstrates the effectiveness of naturally occurring molecules in the inhibition of albumin fibril formation.
Assuntos
Albumina Sérica/metabolismo , Tiroxina/análogos & derivados , Tri-Iodotironina/análogos & derivados , Amiloide/metabolismo , Animais , Bovinos , Relação Dose-Resposta a Droga , Desenho de Fármacos , Temperatura Alta , Cinética , Ligação Proteica/efeitos dos fármacos , Estrutura Secundária de Proteína/efeitos dos fármacos , Albumina Sérica/química , Espectroscopia de Infravermelho com Transformada de Fourier , Tiroxina/farmacologia , Tri-Iodotironina/farmacologiaRESUMO
Este estudo avaliou o seguimento de pacientes com câncer de tireóide de alto risco, após a terapia inicial. Foram selecionados 125 pacientes de alto risco (tumor >4 cm e/ou invasão extra-tireoidiana e/ou metástases linfonodais e idade >45 anos), com ressecção tumoral completa. Todos foram tratados com tireoidectomia total e ablação com 131I [3,75,5 GBq (100150 mCi)] e foram excluídos 18 casos (14,8 por cento) com metástases na PCI pós-dose (t-PCI). O valor preditivo negativo da Tg estimulada <1 ng/ml combinada ao US cervical na primeira avaliação (612 meses após a terapia ablativa) foi de 96,2 por cento para ausência de recidivas em até 5 anos. Este valor aumentou para 98,7 por cento quando acrescentamos a PCI com 185 MBq (5 mCi) 131I (d-PCI). O valor preditivo positivo (VPP) da Tg estimulada >1 ng/ml foi de 52 por cento para presença de metástases detectadas até 5 anos depois, mas considerando apenas pacientes que apresentaram d-PCI e US negativos inicialmente, o VPP foi 19 por cento (9 por cento se Tg 110 ng/ml vs. 40 por cento se Tg >10 ng/ml). Tg reduziu espontaneamente nos pacientes com Tg estimulada >1 ng/ml na primeira avaliação, US e d-PCI negativos e sem recidiva no seguimento, sendo indetectável em metade destes ao final de 5 anos. No primeiro ano após a terapia ablativa, 20 pacientes tiveram captação em leito tireoidiano na d-PCI com Tg estimulada e US negativos e não foram tratados com 131I; estes evoluíram sem recidiva e 60 por cento apresentavam uma d-PCI sem nenhuma captação após 5 anos. Em pacientes sem doença aparente (no US e d-PCI) e Tg estimulada <1 ng/ml, a recidiva em 5 anos foi de apenas 1,3 por cento. Um algoritmo para o seguimento de pacientes de alto risco após a terapia inicial é apresentado por este estudo.
This study evaluated the follow-up of high-risk patients with thyroid cancer after initial therapy. A total of 125 high-risk patients (tumor >4 cm and/or extrathyroid invasion and/or lymph node metastases, and age >45 years), with complete resection of the tumor, were selected. All patients underwent total thyroidectomy and ablation with 131I [3.75.5 GBq (100-150 mCi)]. Eighteen patients (14.8 percent) presenting metastases on post-dose whole-body scan (RxWBS) were excluded. The negative predictive value of stimulated Tg <1 ng/ml in combination with neck US during first assessment (612 mo. after ablative therapy) was 96.2 percent for the absence of recurrence up to 5 years. This value increased to 98.7 percent when adding WBS performed with 185 MBq (5 mCi) 131I (DxWBS). The positive predictive value (PPV) of stimulated Tg >1 ng/ml was 52 percent for the detection of the presence of metastases up to 5 years; however, considering only patients with initially negative DxWBS and US, the PPV was 19 percent (9 percent if Tg of 110 ng/ml vs. 40 percent if Tg >10 ng/ml). Tg levels decreased spontaneously in patients with stimulated Tg >1 ng/ml during first assessment, negative US and DxWBS, and no recurrence during follow-up, with Tg being undetectable in half these patients at the end of 5 years. Twenty patients presented uptake in the thyroid bed upon DxWBS during the first year after ablative therapy, with stimulated Tg and US being negative, and were not treated with 131I; these patients did not relapse and no uptake on DxWBS was observed in 60 percent after 5 years. Recurrence after 5 years was only 1.3 percent in patients without apparent disease (negative US and DxWBS) and stimulated Tg <1 ng/ml. An algorithm for the follow-up of high-risk patients after initial therapy is presented in this study.