RESUMO
BACKGROUND: Leptin, the product of ob gene, is an important circulating hormone for the regulation of homeostasis of body weight and enegy expenditure. There was a previous reports that thyroid hormone is one of regulating factors of leptin gene expression in vitro. The aim of this study was designed to evaluate the role of thyroid hormone levels in the regulation of circulating leptin concentrations in human. METHODS: A total 16S subjects were studied; 76 patients with Graves disease, 49 patients with Hashimoto disease and 43 control sujjects. The correlation between thryoid hormone and leptin levels were analyzed and serum leptin levels were compared among the groups which was classified by thyroid functional status. Serum leptin concentratios were measured by radioimmunoassay. RESULTS: There were no significant differences in serum leptin levels between the groups of control, Graves disease and Hashimoto disease. The hypothyroid groups of Graves disease which was induced by excessive antithyroid drug treatment showed significant low levels(5.6 +/-2.8 ng/mL) compared to control(9.6 +/- 5.2 ng/ml) and thyrotoxic groups(10.0 +/- 5,0 ng/mL) CONCLUSION: The hypothyroid patients showed low levels of serum leptin concentrations it may indicate that thyroid horrnone play a role in the appropriate secretion of leptin in human.
Assuntos
Humanos , Peso Corporal , Expressão Gênica , Doença de Graves , Doença de Hashimoto , Gastos em Saúde , Homeostase , Leptina , Radioimunoensaio , Glândula TireoideRESUMO
The Changes of soluble Fas levels in Patients with Autoimmune Thyroid Diseases BACKGROUD: Apoptosis was observed in thyroid tissue from Hashimoto disease but not those from Graves disease. Recently Fas and Fas ligand interactions among thyrocytes were suggested to development of clinical hypothyroidism in Hashimoto disease.Soluble Fas produced as the form lacking the tranmembrane domain due to alternative splicing, is supposed to inhibit Fas-Fas ligand interaction and blocks Fas mediated apoptosis. METHODS: In tbis study, we measured serum soluble Fas to determine the possible involvement of this molecule in the autoimmune thyroid disease by enzyme linked immunosorbant assay in 29 patients with Graves disease, 30 patients with Hashimotos disease and 19 normal controls. RESULTS: Compared with normal subjeets (4.26 +/- 1.00 U/mL), soluble Fas was not increased in patients with Graves disease (4.23 +/- 1.14 U/mL, p>0.05) but it was increased in throtoxic Graves patients (4.70 +/- 1.26 U/mL, p<0.05) compared to euthyroid Graves (3.72 +/- 0.73 U/mL, p<0.05) and normal subjects (4.26 +/- 1.00 U/mL, p<0.05). The euthyroid and hypothyroid patients with Hashimoto disease showed low soluble Fas levels, 2.94 +/- 0.54 U/mL and 2.74 U/mL, respectively compare to the patients with Graves disease and normal subjects. The thyroid hormone levels to (T3 T4 and free T4) showed positive correlation with the serum titers of antithyroid autoantibodies, antithyroglobuin antibodies, antiperoxidase antibodies and thyrotropin binding inhibitor immunoglobulins. CONCLUSION: We found that the patients with thyrotoxic Graves disease had increased level of serum soluble Fas and the patients with Hashimoto disease showed low levels of soluble Fas compared to normal controls. Increased soluble Fas in Graves disease suggests increased expression of alternatively spliced Fas mRNA variant and decreased soluble Fas in Hashimoto disease suggests decreased Fas mRNA variant and increased full length membrane Fas, so these findings are related to the promotion of apoptosis of thyroid cells during autoimmune reaction in Hashimotos disease.
Assuntos
Humanos , Processamento Alternativo , Anticorpos , Apoptose , Autoanticorpos , Proteína Ligante Fas , Doença de Graves , Doença de Hashimoto , Hipotireoidismo , Imunoglobulinas , Membranas , RNA Mensageiro , Doenças da Glândula Tireoide , Glândula Tireoide , TireotropinaRESUMO
BACKGROUND: The proinflammatory cytokine, IFN-y has been shown to exert pleiotropic effects in a variety of pathophysiologic conditions in autoimmune thyroid disease. The thyrocyte response to IFN-y is mediated two distinct classes of proteins, Janus kinases(Jakl and Jak2) and Signal Transducers and Activation of Transcription(STATl). The activation of STAT 1 is involved in the regulation of many interferon stimulated genes, such as MHC class II, intercellular adhesion molecules-1(ICAM-1) and MHC class II transactivator(CIITA) after the binding to the GASgFN- pactivated site) of the gene promoters. Recently we found TSH/forskolin inhibits IFN-y stimulated maximal expression of ICAM-1 in FRTL-5 cell. IFN-y action is localized between -175 bp and -97 bp from the start of translation of ICAM-1 gene which contains regulatory elements known to be involved in IFN-y action in other eukaryotic cells, palindromic IFN-y activated site(GAS)(5-TTTCCGGGAAA-3) which could bind STAT1, STAT3, STAT5, STAT6. Furthermore, the addition of TSH and forskolin causes a decrease in ICAM-1 promoter activity and its action was localized in GAS. These findings suggested TSH/cAMP signaling pathways downregulate IFN-y activated Janus kinase-STAT signaling path. We wanted to explore the possible involvement of elevated cAMP in the negative regulation of IFN-y induced STAT1 activation in thyroid cells. METHOD: We made several 5-deletion constructs of rat ICAM-1 promoter and analyzed the promoter activities by measuring the luciferase activity after tranfection into FRTL-5 cells. The protein/DNA complex was measured by electrophoretic mobility shift analysis using labeled oligonucleotide. We checked the level of total and phosphorylated STATl protein by immunoblot analysis using specific antibodies. RESULTS: Stimulation of IFN-y in FRTL-5 cells resulted in rapid activation of STATl/DNA binding activity, which was apparent after several minute of stimulation, maintains its activity until 48 h. Incubation of cells with TSH result in suppression of IFN-p mediated STAT1/DNA binding activity throughout the time course of activation by IFN-y. Addition of TSH into 5H maintained FRTL-5 cells did not change the total amount of latent STAT1 amount and also not affect IFN-y mediated production of total STAT1 until 4 h. IFN-y(100 U/mL) rapidly induced phosphorylation of STAT1 within 30 min. and maintained its level without significant change until 48 hours. Cells treated with TSH dramatically lowered the level of IFN-y induced production and phosphorylation of STAT1 after 12 h, 24 h, 36 h, and 48 h but TSH had no effect on the level of phosphorylated STATl within 4 h after IFN-y stimulation. The proteasome inhibitor, MG132 and phosphatase inhibitor, sodium orthovanadate did not block the TSH or forskolin mediated downregulation of phosphorylated STAT1. CONCLUSION: These results indicate a regulatory mechanism which TSH signaling can modulate the prolonged activation of Jak/Stat by IFN-y. We identified one of mechanisms related to TSH mediated negative suppression of the ICAM-1 gene; TSH/cAMP signaling pathways downregulate the cytokine activated Janus kinase-STAT signaling path.
Assuntos
Animais , Ratos , Anticorpos , Colforsina , Regulação para Baixo , Células Eucarióticas , Expressão Gênica , Molécula 1 de Adesão Intercelular , Interferons , Luciferases , Fosforilação , Inibidores de Proteassoma , Sódio , Doenças da Glândula Tireoide , Glândula Tireoide , Tireotropina , Transdutores , VanadatosRESUMO
BACKGROUND: We have found abnormal expression of ICAM-1 in thyroid follicular cells from patients with Graves disease and Hashimoto disease. In this report, we present the hormonal regulation of ICAM-1 mRNA expression and the primary structure of 5-regulatory region which is important for transcriptional regulation of ICAM-1 gene. A I.S kb fragment of the 5-regulatory sequences are identified and linked to luciferase as a reporter. METHOD: Those reporter constructs were used to evaluate the expression in response to cytokines and hormones. Deletion analysis of 1.8 kb fragment of ICAM-1 promoter in FRTL-5 cells provide the evidence for the existence of several regulatory elements of enhancer and silencer in ICAM-1 gene transcription in thyroid cells. RESULTS: ICAM-1 mRNA is easily detected by Northern analysis using total RNA from FRTL-5 cells regardless of culture conditions. The transcripts of rat ICAM-1 showed single band of 2.6 kb in length. The FRT cells which was come from early FRTL cell culture did not show ICAM-1 mRNA with usual Northern analysis, We found differential regulation of ICAM-1 RNA level in different culture condition in FRTL-5 cells, The cells maintained at 3H (no hydrocortisone, no insulin, no TSH) condition showed the highest expression level compared to 4H, 5H, or 6H medium. Hydrocortisone markedly decreased the ICAM-1 RNA and insulin partially recovered the hydrocortisone induced repression. TSH which is important in growth and function of FRTL-5 cells could independently downregulate the ICAM-1 RNA levels. Forskolin (10 mM) could mimic the action of TSH on ICAM-1 mRNA. TNF-a and interferon-y increase ICAM-1 expression in FRTL-5 thyroid cells. TSH/forskolin inhibited maximal expression of ICAM-1 by TNF-a and interferon-r. Promoter activity of the ICAM-1 gene was positively regulated by cytokines, TNF-a and IFN-r and negatively regulated by thyroid stimulating hormone. The addition of TSH and FSK caused a 50% decrease in ICAM-1 promoter activity within 24 hour. The TSH and FSK action was mapped at 175 bp and 97 bp of the start of translation. The mutant construct pCAM-175 delGAS which has no GAS sequence showed no TSH mediated suppression of promoter activity. CONCLUSION: These findings suggested that hormones and cytokines differentially regulated the ICAM-1 gene expression and TSH downregulated ICAM-1 gene transcription by inhibiting the activation of IFN-r induced transcription factors which can bind the GAS of ICAM-1 promoter.