ウシ副甲状腺単層培養細胞を用いた副甲状腺ホルモンに関する研究 : 副甲状腺ホルモンの分泌に対する諸種ビタミンD誘導体の影響
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In order to study the effects of 1, 25-dihydroxyvitamin D<SUB>3</SUB> [1, 25- (OH) <SUB>2</SUB>D<SUB>3</SUB>], 24, 25- dihydroxyvitamin D<SUB>3</SUB> [24, 25 (OH) <SUB>2</SUB>D<SUB>3</SUB>], 1α-hydroxyvitamin D<SUB>3</SUB> (1α-OH-D<SUB>3</SUB>) and 25- hydroxyvitamin D<SUB>3</SUB> (25-OH-D<SUB>3</SUB>) on the secretion of the parathyroid hormone (PTH), monolayer cultures of bovine parathyroid cells already reported in this Journal were used.<BR>Monolayer bovine parathyroid cells cultured for 5 days were preincubated in 1% BSA·MEM·Hanks·HEPES (15 mM) buffer for 2 hr at 37°C, and then incubated in the fresh buffer containing various concentrations of calcium, 10<SUP>-8</SUP>-10<SUP>-10</SUP>M 1, 25 (OH) <SUB>2</SUB>D<SUB>3</SUB>, 10<SUP>-7</SUP> 10<SUP>-9</SUP>M 24, 25 (OH) <SUB>2</SUB>D<SUB>3</SUB>, 10<SUP>-5</SUP>-10<SUP>-7</SUP>M 1α-OH-D<SUB>3</SUB> and/or 10<SUP>-5</SUP>-10<SUP>-7</SUP>M 25-OH-D<SUB>3</SUB> for up to 12 hr. In some experiments, 10 mM theophylline was included in the medium, or 10<SUP>-6</SUP>M prostaglandin (PG) E<SUB>2</SUB> was added 30 min before the termination of the experiment. PTH concentrations in the incubation medium were determined by radioimmunoassay. Cell cyclic AMP (cAMP) and cyclic GMP (cGMP) contents were determined through competitive protein binding assay and radioimmunoassay, respectively.<BR>In a preliminary study to evaluate the viability of the monolayer cultures of bovine parathyroid cells incubated up to 12 hr, PTH secretion rates increased almost lineally as a function of incubation time in a low calcium medium (0.5 mM), remained almost constant in a normal calcium medium (1.1 mM) and decreased in a high calcium medium (4.4 mM). When the monolayer bovine parathyroid cells were incubated in a low calcium medium containing 10<SUP>-8</SUP>M 1, 25 (OH) <SUB>2</SUB>D<SUB>3</SUB>, the PTH secretion rate increased slightly for the first 4 hr followed by a decrease thereafter. All the cells incubated up to 12 hr were viable, as assessed by the trypan blue dye exclusion method.<BR>10<SUP>-8</SUP>-10<SUP>-10</SUP>M 1, 25 (OH) <SUB>2</SUB> D<SUB>3</SUB> and/or 10<SUP>-7</SUP>M 24, 25 (OH) <SUB>2</SUB> D<SUB>3</SUB> significantly decreased the PTH secretion rate compared to that of the control group, when monolayer bovine parathyroid cells were incubated with these substances in low or normal calcium mediums. An increase of the PTH secretion rate by 10<SUP>-6</SUP>M PGE<SUB>2</SUB> or 10 mM theophylline was also inhibited through 10<SUP>-8</SUP>M 1, 25 (OH) <SUB>2</SUB> D<SUB>3</SUB>. Neither 10<SUP>-5</SUP>-10-7M 1α-OH-D<SUB>3</SUB> nor 10<SUP>-5</SUP>-10<SUP>-7</SUP>M 25-OH-D<SUB>3</SUB> showed any effects on the PTH secretion rate and cell cAMP and cGMP contents.
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