The activity of NADH-cytochrome b(5) reductase (b(5)R) and the levels of hydrogen peroxide in the thyroid of patients with Graves' disease (GD) and normal controls were measured with potassium ferricyanide as substrate and with the homovanillic acid fluorescence assay. The activity of b(5)R and the level of H2O2 in GD thyroid were definitely higher than those in normal control, but the activity of catalase in GD thyroid was not significantly different from that in normal thyroid. After addition of p-chloromercuribenzoate, a b(5)R inhibitor, the activity of b(5)R in GD and normal thyroid decreased by 85%, the level of H2O2 decreased by 50%, and protein-bound iodine (PBI) formation by 52%. There is a positive correlation between the level of H2O2 and the activity of b(5)R. Our data indicate that b(5)R participates in thyroid hydrogen peroxide biosynthesis, and is an important enzyme in the production of H2O2.
Case-Control Studies ; Cytochrome-B(5) Reductase ; metabolism ; Graves Disease ; metabolism ; Humans ; Hydrogen Peroxide ; metabolism ; Thyroid Gland ; enzymology

BACKGROUNDType 1 deiodinase (D1) plays an important role in the metabolism of thyroid hormone and has close relationship with thyroid function. In this study we explore the effects of iodine intake on D1 activity and its mRNA expression and its possible mechanism.

METHODSForty-eight Wistar rats were randomly divided into six groups with 8 in each: low iodine (LI), normal iodine (NI), five-fold iodine (HI(5)), ten-fold iodine (HI(10)), fifty-fold iodine (HI(50)), one hundred-fold iodine (HI(100)) group. Three months, six months and twelve months after admistration of potassium iodate, they were sacrificed and thyroids were excised. The expression of D1 mRNA in the thyroid tissue was determined by RT-PCR and D1 activity was analyzed by (125)I-rT3 as substrate. The thyroid hormone was measured with radioimmunoassay method.

RESULTSCompared with NI group, D1 mRNA expression in LI groups slightly decreased, and D1 activity greatly increased. Both T(3) and T(4) in thyroid tissue significantly decreased, but the T(3)/T(4) ratio increased. D1 mRNA expression decreased in all HI groups, and D1 activity was significantly lower in HI groups. There was a tendency of decrease in D1 activity with increased doses of iodine intakes. There was no significant difference in T(4) in thyroid tissue between HI groups and NI group, but a tendency of decrease in T(3) level was found in all HI groups.

CONCLUSIONSIn the case of iodine deficiency, D1 activity increased greatly in order to convert more T(4) to T(3). Excess iodine can inhibit both D1 mRNA expression and its activity to protect organism from being injured by excessive T(3).

Animals ; Iodide Peroxidase ; genetics ; metabolism ; Iodine ; administration & dosage ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Thyroid Gland ; enzymology ; Thyroxine ; blood ; Triiodothyronine ; blood

Ceramide, a product of sphingomyelin hydrolysis, is now recognized as an intracellular lipid messenger, which mediates the effects of extracellular agents on cellular growth, differentiation and apoptosis. Recently, ceramide has been implicated in the regulation of phospholipase D (PLD). In this study, we examined the effects of ceramide on the activity and mRNA level of PLD during apoptotic process in FRTL-5 thyroid cells. C2-ceramide (N-acetyl sphingosine) induced apoptosis in FRTL-5 thyroid cells. Fluorescent staining showed that ceramide induced the typical features of apoptosis including condensed or fragmented nuclei. DNA fragmentation was also observed by agarose gel electrophoresis. Flow cytometric cell cycle analysis showed more clearly that ceramide induced apoptotic cell death in FRTL-5 thyroid cells. The treatment of FRTL-5 thyroid cells with thyroid-stimulating hormone (TSH) resulted in an increased PLD activity in a dose- and time-dependent manner. However, the TSH-induced increase in PLD activity was down-regulated within 2 h after ceramide treatment. Furthermore, the levels of PLD mRNA were found to be decreased throughout apoptotic process as inferred by reverse transcription-polymerase chain reaction. However, the decreases in PLD mRNA levels were not correlated with those in PLD activities after ceramide treatment. Taken together, these data suggest that ceramide inhibits the PLD activity in an early apoptotic phase and down-regulation of the levels of PLD mRNA may be implicated in apoptotic process in FRTL-5 thyroid cells.
Animal ; Apoptosis/drug effects* ; Cells, Cultured ; DNA Fragmentation ; Enzyme Activation/drug effects ; Flow Cytometry ; Gene Expression Regulation, Enzymologic/drug effects ; Phospholipase D/metabolism* ; Phospholipase D/genetics ; RNA, Messenger/genetics ; Rats ; Rats, Inbred Strains ; Sphingosine/pharmacology ; Sphingosine/analogs & derivatives* ; Thyroid Gland/enzymology ; Thyroid Gland/drug effects* ; Thyrotropin/pharmacology