Resistance to thyroid hormone (RTH) is an autosomal dominant disorder that's characterized by inappropriate normal or elevated TSH levels despite of the elevated thyroid hormone levels. RTH is distinguished from the TSH secreting pituitary adenoma by performing the TRH stimulation test, TSH alpha subunit measurement and sellar MRI. A 23 year old woman visited our hospital complaining of fatigue, palpitation and heat intolerance and she had an anterior neck mass. She had elevated total T3, free T4 and TSH levels. The serum TSH levels were increased during the TRH stimulation test before and after T3 suppression. The serum TSH alpha subunit showed a normal response and the serum TSH alpha subunit/TSH molar ratio did not increase over 1.0 with TRH stimulation. Thyroid hormone receptor beta gene mutation was identified. Although a left pituitary microadenoma was revealed on sellar MRI, the patient was diagnosed as having pituitary RTH with a nonfunctioning pituitary microadenoma. We report here on a patient with pituitary RTH and a nonfunctioning pituitary microadenoma, and this is the first such case in Korea.
Fatigue ; Female ; Glycoprotein Hormones, alpha Subunit ; Hot Temperature ; Humans ; Molar ; Neck ; Pituitary Neoplasms ; Thyroid Gland ; Thyroid Hormone Receptors beta

OBJECTIVETo study the interaction between insulin-like growth factor binding protein-3 (IGFBP-3) and thyroid hormone receptor α1 (TRα1) and the modulatory effect of IGFBP-3 on transcription of the thyroid hormone responsive gene.

METHODSThe interaction between IGFBP-3 and TRα1 was detected with glutathione-S-transferase pull-down method, co-immunoprecipitation, fluorescence resonance energy transfer test. The cellular distribution of these two proteins was observed by confocal laser scanning microscopy. The effect of IGFBP-3 on the growth hormone promoter activity stimulated by triiodothyronine (T3) was determined by dual-luciferase reporter assay.

RESULTSIGFBP-3 interacted with TR&alpha;1 both in vivo and in vitro. IGFBP-3 and TR&alpha;1 were shown to co-localize in the nucleus of HEK-293 cells. The overexpressed IGFBP-3 inhibited the growth hormone promoter activity stimulated by T3 (P<0.01).

CONCLUSIONSIGFBP-3 interacts with TR&alpha;1 and inhibits T3 responsive gene transcription. This finding further confirms the insulin-like growth factor-independent role of IGFBP-3 in the nucleus.

HEK293 Cells ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; metabolism ; Promoter Regions, Genetic ; Thyroid Hormone Receptors alpha ; metabolism ; Thyroid Hormones ; genetics ; metabolism ; Transcription, Genetic ; Triiodothyronine ; pharmacology

OBJECTIVE: To preliminarily investigate the relationship between stimulatory G protein α subunit (GNAS) and thyroid hormone receptor α (THRA) gene mutations and clinical phenotypes in children with congenital hypothyroidism (CH). METHODS: A total of 70 children with CH diagnosed by neonatal screening were enrolled. Their peripheral blood samples were collected to extract genomic DNA. GNAS and THRA genes were screened for mutations using next-generation sequencing. Bioinformatics software was used to analyze the pathogenicity of gene mutations. RESULTS: Of the 70 children with CH, nine missense mutations (three known mutations and six novel mutations) in the GNAS gene were detected in three patients (4%), and one gene polymorphism, c.508A>G(p.I170V), in the THRA gene was detected in four patients. The analysis results of bioinformatics software and ACMG/AMP guidelines showed that the two GNAS gene mutations [c.301C>T(p.R101C) and c.334G>A(p.E112K)] were more likely to be pathogenic. Three children with GNAS gene mutations showed different degrees of hypothyroidism. CONCLUSIONS GNAS gene mutations are related to the development of CH, and children with CH have different clinical manifestations. THRA gene mutations may not be associated with CH.
Chromogranins ; genetics ; Congenital Hypothyroidism ; GTP-Binding Protein alpha Subunits, Gs ; genetics ; Genes, erbA ; Humans ; Infant, Newborn ; Mutation ; Phenotype ; Thyroid Hormone Receptors alpha ; genetics

BACKGROUND: Expression of hepatic cholesterol 7alpha-hydroxylase (CYP7A1) is negatively regulated by orphan nuclear receptor small heterodimer partner (SHP). In this study, we aimed to find whether thyroid hormone regulates SHP expression by modulating the transcriptional activities of liver receptor homolog-1 (LRH-1). METHODS: We injected thyroid hormone (triiodothyronine, T3) to C57BL/6J wild type. RNA was isolated from mouse liver and used for microarray analysis and quantitative real-time polymerase chain reaction (PCR). Human hepatoma cell and primary hepatocytes from mouse liver were used to confirm the effect of T3 in vitro. Promoter assay and electrophoretic mobility-shift assay (EMSA) were also performed using human hepatoma cell line RESULTS: Initial microarray results indicated that SHP expression is markedly decreased in livers of T3 treated mice. We confirmed that T3 repressed SHP expression in the liver of mice as well as in mouse primary hepatocytes and human hepatoma cells by real-time PCR analysis. LRH-1 increased the promoter activity of SHP; however, this increased activity was markedly decreased after thyroid hormone receptor beta/retinoid X receptor alpha/T3 administration. EMSA revealed that T3 inhibits specific LRH-1 DNA binding. CONCLUSION: We found that thyroid hormone regulates the expression of SHP mRNA through interference with the transcription factor, LRH-1.
Animals ; Bile Acids and Salts ; Carcinoma, Hepatocellular ; Cell Line ; Child ; Child, Orphaned ; Cholesterol ; Cholesterol 7-alpha-Hydroxylase ; DNA ; Hepatocytes ; Humans ; Liver* ; Mice ; Microarray Analysis ; Real-Time Polymerase Chain Reaction ; Receptors, Thyroid Hormone ; RNA ; RNA, Messenger* ; Thyroid Gland* ; Thyroid Hormones ; Transcription Factors

To study the role of the thyroid hormone receptor TRalphaA involved in the process of the metamorphic development of Japanese flounder, we firstly cloned the TRalphaA gene, then ligated into the fusion expression vector pET30a and expressed in Escherichia coli DE3 (BL21) host cells. After induced for 4 h with 1 mmol/L Isopropyl beta-D-Thiogalactoside, the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. The recombinant protein was denatured and purified by His-Bind resin, then renatured through gradient washing on His-bind resin column. After that, polyclonal antibody was prepared by immunizing New Zealand rabbits with purified protein. Dot blotting analysis showed the antibody with the titer of 1:200 000 reacted specifically to the expressed recombinant protein. Furthermore, a chromatin immunoprecipitation assay was performed to identify the specific binding between the antibody and TRalphaA in living cells of Japanese flounder. The result showed that thyroid hormone was involved in the alkaline phosphatase (ALP) gene transcriptional regulation through TRalphaA in vivo.
Alkaline Phosphatase ; genetics ; immunology ; Animals ; Antibodies ; immunology ; Escherichia coli ; genetics ; metabolism ; Flounder ; physiology ; Metamorphosis, Biological ; physiology ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Thyroid Hormone Receptors alpha ; biosynthesis ; genetics ; immunology

OBJECTIVETo investigate the relationship of NOR-1 with the inhibition of inflammatory reaction in mice Kupffer cells (KCs) induced by lipopolysaccharide (LPS) via liver X receptor alpha (LXR alpha).

METHODSKCs from male KM mice were isolated by density gradient centrifugation, incubated and then randomly assigned to three groups: control group, LPS treated group and LPS+T0901317 treated group.

RESULTSThe mRNA and protein expressions of LXR alpha and NOR-1 in each group were determined by RT-PCR, immunofluorescent assay and western blot, respectively. The densities of TNF alpha and IL-10 in supernatants were evaluated by enzyme linked immunosorbent assay (ELISA). The mRNA and protein expression levels of LXR alpha in LPS + T0901317 group were the highest as compared to the other two groups (0.748+/-0.072 and 1.217+/-0.133 respectively), The mRNA and protein expression levels of NOR-1 in LPS+ T0901317 group were the highest as compared to the other two groups (2.726+/-0.065 and 0.842+/-0.058 respectively). The densities of supernatant TNF alpha in LPS group and IL-10 in LPS+T0901317 group were the highest [(450.89+/-78.52) ng/L and (537.41+/-36.41) ng/L respectively].

CONCLUSIONSPromoting the expression of LXR alpha in KCs can elevate the NOR-1 expression and then inhibit inflammatory reaction.

Animals ; Cells, Cultured ; DNA-Binding Proteins ; metabolism ; Inflammation ; metabolism ; Interleukin-10 ; metabolism ; Kupffer Cells ; metabolism ; Liver X Receptors ; Male ; Mice ; Mice, Inbred Strains ; Nerve Tissue Proteins ; metabolism ; Orphan Nuclear Receptors ; metabolism ; Receptors, Steroid ; metabolism ; Receptors, Thyroid Hormone ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism