Comparison of the relative effects of 1,24-dihydroxyvitamin D(2) [1, 24-(OH)(2)D(2)], 1,24-dihydroxyvitamin D(3) [1,24-(OH)(2)D(3)], and 1,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] on selected vitamin D-regulated events in the rat

The present experiments were conducted to compare the relative hypercalciuric and hypercalcemic activities of 1,24-dihydroxyvitamin D(2) [1,24-(OH)(2)D(2)], 1,24-dihydroxyvitamin D(3) [1, 24-(OH)(2)D(3)], and 1,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] in 7-week-old rats. The rats were dosed orally with each sterol for 7 days at a rate of 1 ng/g body weight/day. We also monitored the effect of the three compounds on the induction of mRNA for CaATPase and for 25-hydroxyvitamin D-24-hydroxylase in the kidney and intestine, on plasma vitamin D metabolite levels, and on the capacity to evoke modification in the vitamin D receptor/retinoic acid X receptor (VDR/RXR) heterodimer conformation. Plasma calcium was elevated in the rats treated with 1,24-(OH)(2)D(3) and 1, 25-(OH)(2)D(3), but not in the 1,24-(OH)(2)D(2)-dosed rats. Urinary calcium was elevated significantly (relative to controls) in all groups. The order of hypercalciuric activity was 1,25-(OH)(2)D(3) >/= 1,24-(OH)(2)D(3) >/= 1,24-(OH)(2)D(2) > control. Duodenal plasma membrane calcium ATPase (PMCA) mRNA was elevated to a similar extent in all groups relative to controls. Duodenal 24-hydroxylase mRNA was elevated in all groups relative to controls; however, the elevations were significantly higher in the 1,24-(OH)(2)D(3) and 1, 25-(OH)(2)D(3) groups compared with the 1,24-(OH)(2)D(2) group. Kidney 24-hydroxylase also was elevated significantly in the 1, 24-(OH)(2)D(3)- and 1,25-(OH)(2)D(3)-treated rats but not in the 1, 24-(OH)(2)D(2)-treated rats. Recombinant human vitamin D receptor (hVDR) extracts were incubated with saturating concentrations of 1, 24-(OH)(2)D(2), 1,24-(OH)(2)D(3), and 1,25-(OH)(2)D(3) and subsequently analyzed by electrophoretic mobility shift assay (EMSA). Overall binding was comparable for all metabolites; however, the 1, 24-(OH)(2)D(2) complex exhibited distinctly altered mobility relative to 1,24-(OH)(2)D(3) and 1,25-(OH)(2)D(3), suggestive of an effect on hVDR/hRXR conformation. These data suggest that 1, 24-(OH)(2)D(2) is not as potent as either of the vitamin D(3) sterols at affecting hypercalcemia or hypercalciuria in young growing rats; however, 1,24-(OH)(2)D(2) can evoke other biological responses similar to the vitamin D(3) sterols. These different responses may be related to the alterations in conformation state of the hVDR/hRXR heterodimer.     

Calcipotriol (MC 903): pharmacokinetics in rats and biological activities of metabolites. A comparative study with 1,25(OH)2D3

Calcipotriol (MC 903) is a novel analogue of the physiologically active metabolite of vitamin D3, 1 alpha,25-dihydroxycholecalciferol [1,25(OH)2D3]. MC 903 and 1,25(OH)2D3 have similar effects on cell proliferation and cell differentiation in vitro using the human histiocytic lymphoma cell line U 937, but in vivo MC903 has 100-200 times less effect on calcium metabolism. To elucidate this difference, the pharmacokinetic profiles after a single intravenous dose (50 micrograms/kg) of the two compounds to rats were compared. The area under the serum level/time curve (AUC) was more than 100 times higher for 1,25(OH)2D3 than for MC903 and the rate of clearance was more than 100 times higher for MC903 than for 1,25(OH)D3. Serum from MC903 or 1,25(OH)2D3 dosed rats (i.v. 10 micrograms/kg) was investigated for biological activities by incubation of U 937 cells with serum collected 0-24 hr after drug administration. Serum from MC903 dosed rats had an effect only when collected shortly after dosing, whereas serum from 1,25(OH)2D3 dosed rats had an effect when collected up to 4 hr after dosing. The biological effects on the U937 cells of the two major metabolites of MC903 (MC 1046 and MC 1080) were investigated. The metabolites had effects that were more than 100 times weaker than those of the parent compound. The effect of MC903 on proliferative disorders, its fast elimination and the formation of inactive metabolites makes MC903 suitable for topical treatment of psoriasis.     

Influence of cadmium on the metabolism of vitamin D3 in rats

In order to obtain further information on the effects of cadmium (Cd) on the mechanism of activation of vitamin D3 in the kidney, the serum concentrations of 1,25-dihydroxycholecalciferol [1,25(OH)2D3] and 24,25-dihydroxycholecalciferol [24,25(OH)2D3] were determined by radioimmunoassay techniques. The serum concentration of 1,25(OH)2D3 in Cd-exposed rats was always higher than that in control rats. The concentrations of serum 1,25(OH)2D3 in parathyroidectomized rats (PTX), both in the control and in the Cd-exposed groups, were markedly lower than those in non-PTX rats. On the other hand, the concentration of serum 24,25(OH)2D3 in Cd-exposed rats was less than that in control rats. In other words, the pathway from secretion of parathyroid hormone (PTH) to synthesis of 1,25(OH)2D3 or 24,25(OH)2D3 was normal, but the pathway from stimulation of PTH to secretion of PTH was abnormal. These results suggest that rats exposed to Cd for 90 days were in a state of either deficiency of 1,25(OH)2D3 or excess secretion of parathyroid hormone. Although hypophosphatemia occurred in the Cd-exposed rats, the 1,25(OH)2D3 serum level in rats was not increased by PTX. On the basis of these results, it is suggested that hypophosphatemia occurring after the exposure of rats to Cd is a secondary hypophosphatemia.     

Regulation of sodium, calcium and vitamin D metabolism in Dahl rats on a high-salt/low-potassium diet: genetic and neural influences

1. A dietary combination of high salt and low potassium (HSLK) exacerbates hypertension in Dahl salt-sensitive (DS) rats and renders previously normotensive Dahl salt-resistant (DR) rats hypertensive. In both strains, the severity of hypertension correlates with urinary calcium loss. However, the magnitude of excretory calcium losses is significantly greater in DS rats and is potentiated by chemical sympathectomy in both strains. 2. We hypothesized that a defect in vitamin D metabolism may underlie the observed strain-dependent differences in calcium balance. 3. Arterial blood pressure (ABP), water and mineral balance and serum concentrations of 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3) and 25-hydroxyvitamin D3 (25(OH)D3) were measured in intact and chemically sympathectomized (6-hydroxydopamine; 6-OHDA) DS and DR rats after 8 weeks on a HSLK diet. 4. Chronic ingestion of this diet resulted in marked and moderate levels of hypertension in DS and DR rats, respectively. The hypertension was abated and eliminated by 6-OHDA in the DS and DR strains, respectively. Independent of treatment, DS rats had significantly higher urinary excretion of calcium and reduced intestinal absorption of the ion compared with DR rats. The DS rats had significantly higher serum levels of 1,25(OH)2 D3 and markedly lower serum levels of 25(OH)D3 than DR rats. Chemical sympathectomy tended to increase 1,25(OH)2 D3 and to decrease 25(OH)D3 levels in both strains. 5. These data indicate a genetic difference in vitamin D metabolism between DS and DR rats. The abnormally elevated levels of 1,25(OH)2 D3 in DS rats may be an appropriate compensatory response to excessive excretory calcium loss and reduced target organ sensitivity to the hormone and may, maladaptively, directly contribute to hypertension, by stimulating vascular smooth muscle contractility.     

In vivo and in vitro pharmacokinetics and metabolism studies of 26,26,26,27,27,27-F6-1,25(OH)2 vitamin D3 (Falecalcitriol) in rat: induction of vitamin D3-24-hydroxylase (CYP24) responsible for 23S-hydroxylation in target tissues and the drop in serum levels.

1. 26,26,26,27,27,27-F6,-1,25(OH)2 vitamin D3, Falecalcitriol, the hexafluorinated analogue of 1,25(OH)2 vitamin D3, has been reported to be several times more potent than the parent compound regarding some vitamin D actions. The reason for enhanced biological activity appears related to F6-1,25(OH)2 vitamin D3 metabolism to F6-1,23S,25(OH)3 vitamin D3, a bioactive 23S-hydroxylated form which is resistant to further metabolism. 2. In the present in vivo studies, the repeated oral administration of [3H]F6-1,25(OH)2 vitamin D3 to rat resulted in a significant reduction of the radioactivity and the F6-1,25(OH)2 vitamin D3 concentrations in serum, especially at the 2 h maximum point after each dosing. Additionally, F6-1,23S,25(OH)3 vitamin D3 in the serum and small intestine was increased by the prior administration of F6-1,25(OH)2 vitamin D3. 3. Further in vitro investigation showed [3H]F6-1,25(OH)2 vitamin D3 to be metabolized to F6-1,23S,25(OH)3 vitamin D3 by kidney and small intestine homogenates of rat, the reaction being increased by the prior administration of F6-1,25(OH)2 vitamin D3. Moreover, this latter treatment was associated with a marked increase of CYP24 mRNA in the small intestine within 4 h after dosing. 4. The results indicate that in vivo metabolism of F6-1,25(OH)2 vitamin D3 to F6-1,23S,25(OH)3 vitamin D3 is catalysed by CYP24, the enzyme being induced by prior substrate exposure.     

Contribution of intestinal calcium absorption to 1,25-dihydroxyvitamin D3-induced calcium action in the total parenteral nutrition rat

To investigate the contribution of intestinal calcium (Ca) absorption to 1,25-dihydroxyvitamin D(3) (1,25(OH) (2)D(3))-induced Ca action, we assessed parameters related to Ca metabolism after a single dosing of 1,25(OH)(2)D (3) in the total parenteral nutrition (TPN) solution or 5% D-mannitol (MAN) solution treatment with rats. Animals were divided into 6 groups (vehicle, 100 microg/kg p.o. and 25 microg/kg i.v.; n=8) in Experiment 1 and 8 groups (vehicle, 1, 10 and 100 microg/kg p.o.; n=6) in Experiment 2 at TPN or MAN solution treatment. In both experiments, the parameters related to Ca metabolisms, urinary Ca and urinary deoxypyridinoline on 0-24 hr or serum Ca, osteocalcin and parathyroid hormone at 24 hr were measured after 1,25(OH)(2)D (3) dosing. 1,25(OH)(2)D(3)-related increased urinary Ca or serum Ca were observed in both experiments. Decrease rates in change of urinary Ca in TPN solution treatment rats were 36.3% (100 microg/kg p.o.) or 47.1% (25 microg/kg i.v.) of MAN solution treatment rats in Experiment 1, and 29.0% (1 microg/kg), 56.2% (10 microg/kg) or 35.3% (100 microg/kg) of MAN solution treatment rats in Experiment 2. Decrease rates in change of serum Ca at 72 hr in TPN solution treatment rats were 57.3% (100 microg/kg p.o.) or 44.5% (25 microg/kg i.v.) of MAN solution treatment rats in Experiment 1, and were 57.0% (100 microg/kg) of MAN solution treatment rats in Experiment 2. There were no differences in the change of serum Ca in the 1,25(OH)(2)D(3) 1 or 10-microg/kg group in Experiment 2. Our results suggest that differences in urinary Ca or serum Ca between MAN solution treatment rats and TPN solution treatment rats express the contribution of intestinal Ca absorption to 1,25(OH)(2)D(3)-induced Ca action in the conditions of the study.     

Acute effects of parathyroid extract on renal vitamin D hydroxylases in Japanese quail.

The dose- and the time-response relationships of parathyroid hormone's modulation of in vitro 1,25-(OH)2D3 and 24,25-(OH)2D3 production in Japanese quail were investigated. 4-week-old female Japanese quail were injected intramuscularly with three different doses (30, 90 and 270 USP units/kg) of parathyroid extract (PTH). 4, 12 and 24 h after the in vivo administration of PTH, kidney homogenates were incubated with tritiated 25-(OH)D3. All three doses of PTH stimulated 1,25-(OH)2D3 and inhibited 24,25-(OH)2D3 production at 12 h relative to the vehicle-treated control group. The responses of the vitamin D hydroxylases were not detectable at 4 h and had returned to control levels by 24 h. Plasma calcium rose significantly at 4 h with the two higher doese of PTH (90 and 270 USP units/mg) but not with the lowest dose. By 12 h the plasma calcium had returned to control levels except at the highest dose of PTH. By 24 h all of the plasma calcium levels had returned to control values. Plasma inorganic phosphate levels were depressed at 4 h by all three PTH doses; they remained depressed at 12 h and returned to control levels by 24 h. It is concluded that the pharmacological effects of PTH on the renal vitamin D hydroxylases are slower in onset and shorter in duration when compared with those of estradiol.     

Is 24,25-dihydroxycholecalciferol a calcium-regulating hormone in man?

Small doses (1-10 microgram daily) of 24,25-dihydroxycholecalciferol (24,25-(OH)2D3), a renal metabolite of vitamin D of uncertain function, increased intestinal absorption of calcium in normal people and in patients with various disorders or mineral metabolism, including anephric subjects. In five of six patients studied, calcium balance increased, but, unlike 1,25-dihydroxycholecalciferol, 24,25-(OH)2D3 did not increase plasma or urinary calcium concentrations. These results suggest that 24,25-(OH)2D3 may be an important regulator of skeletal metabolism in man with potential value as a therapeutic agent.     

Action of 24R, 25 dihydroxyvitamin D3 (24,25 (OH)2D3) on bone in vivo

The administration of 24R,25(OH)2D3 for 2 years led to an increase in bone volume in the rat. Radiographs showed the striking increase in the density of the bones from animals treated with large amounts of 24R,25(OH)2D3. Total bone minerals measured by direct single photon absorptiometry showed a dose-dependent increase of up to approximately 150% that of controls in both the femur and coccyx of the treated animals. Histological observation revealed a thickened cortex with narrowed medullary area, and Villanueva's bone stain indicated the increase of mineralized bone. The result clearly demonstrates that 24,25(OH)2D3, if massive doses are given, has the biological action of increasing bone volume without disturbing the development of bone sizes.     

Relationship Between Circulating Vitamin D3 Metabolites and Prolactin or Growth Hormone Levels in Rat

Abstract: Previous studies (Haug & Gautvik 1985) have demonstrated specific receptors for 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) in a clonal (GH₃) strain of rat pituitary tumour cells. It was discovered that 1,25(OH)₂D₃ affected the production of prolactin and growth hormone in these cells in a calcium dependent manner. These findings were the basis for a hypothesis that vitamin D₃ could be involved in the regulation of pituitary hormones in vivo. To further investigate this contention, female rats were given subcutaneous injections of 1,25(OH)₂D₃, 25-hydroxyvitamin D₃ (25(OH)D₃) or 24,25-dihydroxyvitamin D₃ (24,25(OH)₂D₃) three times a week for up to 12 weeks. Blood samples were withdrawn after 28, 56 and 84 days of treatment and analysed for vitamin D, metabolites, prolactin and growth hormone, and serum ionized (free) and total calcium (Ca). Between treatment group comparisons of serum prolactin and growth hormone levels did not show significant vitamin D₃ induced alterations. However, correlation matrix analyses on all variables revealed that serum level of growth hormone was significantly (P < 0.05) and inversely related to corresponding total Ca. Prolactin, on the other hand, may be subject to a complex regulation by 1,25(OH)₂D₃ and free Ca²⁺.     

1,25(OH)2D3对Thy-1肾炎模型大鼠Ki67、mTOR表达的影响

摘要:目的 研究 1,25-二羟基维生素 D3 [1,25(OH)2D3]对 Thy-1 肾炎模型大鼠 Ki67 和哺乳动物雷帕霉素靶蛋白 (mTOR)表达的影响, 并探讨其机制。方法 90 只清洁级雄性 SD 大鼠随机分为对照组、 模型组、 1,25(OH)2D3组, 每组 30 只。模型组与 1,25(OH)2D3组尾静脉注射抗 Thy-1 单克隆抗体建立肾炎模型, 对照组给予等剂量生理盐水。建模 后, 1,25(OH)2D3组给予 1,25(OH)2D3 0.5 μg/d 灌胃, 连续给药 21 d, 对照组及模型组给予等体积花生油。分别于给药 后第 1、 3、 7、 14、 21 天每组随机处死 6 只大鼠, 处死前 1 d 收集 24 h 尿液进行 24 h 尿蛋白定量; 取各组肾组织标本, 经 HE 和 PAS 染色后进行肾脏病理损害评分, 免疫组化法检测肾组织中 Ki67、 mTOR 表达。结果 模型组和 1,25 (OH)2D3组大鼠在建模后第 1 天尿蛋白水平升高, 模型组第 3 天达高峰, 至第 14 天恢复至正常水平, 1,25(OH)2D3组大 鼠第 1、 3、 7 天的尿蛋白水平均低于模型组 （P < 0.05）。1,25(OH)2D3组大鼠肾组织病理损害程度第 3、 7 天较模型组减 轻（P < 0.05）, Ki67、 mTOR 蛋白表达水平较模型组降低（P < 0.05）。24 h 尿蛋白定量, Ki67 表达水平, mTOR 表达水 平及肾组织病理损害评分彼此间均呈正相关。结论 1,25(OH)2D3可抑制 Thy-1 肾炎模型大鼠肾小球系膜细胞的增 殖, 其作用机制可能与减少Ki67、 mTOR 的表达有关。     

Effects of synthetic vitamin D analogues on breast cancer cell proliferation in vivo and in vitro.

Calcipotriol (MC903) is a novel vitamin D analogue which effects cellular differentiation and proliferation in vitro and has reduced effects on calcium metabolism in vivo. In the present study its in vitro activity was evaluated using the MCF-7 breast cancer cell line, and its effects on calcium metabolism and mammary tumour growth were measured in vivo in adult female rats. Calcipotriol was compared to the natural metabolite of vitamin D3, 1 alpha,25-dihydroxycholecalciferol [1,25(OH)2D3] and its synthetic analogue 1 alpha hydroxycholecalciferol [1 alpha(OH)D3]. Both calcipotriol and 1,25(OH)2D3 produced significant inhibition of MCF-7 cell proliferation at a concentration of 5 x 10(-11) M. Intraperitoneal administration of calcipotriol to normal female rats showed that the analogue was 100-200 times less active than 1,25(OH)2D3 in raising serum calcium concentration and urinary calcium excretion. Anti-tumour activity of the vitamin D analogues was investigated in vivo using the nitrosomethylurea-induced rat mammary tumor model. Rats, maintained on a low calcium diet, were treated with 1 alpha(OH)D3 (0.25 and 1.25 micrograms/kg). Both doses produced a response rate of 25% but hypercalcaemia developed. Treatment with calcipotriol (50 micrograms/kg) of rats maintained on a normal laboratory diet caused inhibition of tumour progression (response rate 17%) without the development of severe hypercalcaemia. This study supports the concept that vitamin D derivatives may inhibit breast cancer cell proliferation in vivo.     

The comparative toxicity of vitamin D metabolites in the weanling mouse

The potential toxicity of vitamin D, alpha-calcidol [1 alpha(OH)D3], and calcitriol [1,25(OH)2D3] was studied by administration of these compounds at three different doses to weanling C57BL/6J mice over a 4-week period. Drug effects on calcium were monitored by serum calcium and urine calcium/creatinine ratio determinations. Tests of renal function included serum creatinine, 24-h urine volume, urinary protein, and glucose excretion, and histological evaluation of renal tissue. At 2 weeks, serum calcium was significantly elevated in animals receiving the higher doses of alpha-calcidol (2.78 +/- 0.25 at 50 ng/kg and 3.45 +/- 0.13 at 250 ng/kg body wt vs 2.14 +/- 0.06 mmol/l in controls, respectively). A similar effect was seen in the urinary calcium/creatinine ratio but serum creatinine remained unchanged. By 4 weeks, all animals receiving alpha-calcidol had significantly higher serum calcium and urinary calcium/creatinine ratios than other groups. Severe nephrocalcinosis was observed in the high-dose alpha-calcidol group only. We conclude that alpha-calcidol is more toxic than calcitriol in the mouse and suggest that the degree of toxicity is correlated to the degree of hypercalcemia and to the vitamin D metabolite used.     

Quantitation of serum 25(OH)D2 and 25(OH)D3 concentrations by liquid chromatography tandem mass spectrometry in patients with diabetes mellitus

Vitamin D has been considered to regulate calcium and phosphorus homeostasis and to preserve skeletal integrity. Serum 25-hydroxyvitamin D (25(OH)D) is the best indicator of vitamin D levels. The association of serum 25(OH)D deficiency with increased risk of type 1 diabetes (T1DM) and type 2 diabetes (T2DM) is controversial. We investigated serum 25(OH)D₂ and 25(OH)D₃ levels in diabetes patients by using liquid chromatography tandem mass spectrometry (LC-MS/MS). Serum 25(OH)D2 and 25(OH)D3 levels were measured with liquid chromatography tandem mass spectrometry in electrospray ionization positive mode. Chromatograms were separated using an ACE5 C18 column on a gradient of methanol. The total 25(OH)D levels were calculated as the sum of 25(OH)D3 and 25(OH)D2 levels. A total of 56 patients with T1DM and 41 patients with T2DM were enrolled in this study. There were 42 and 28 non-diabetic, age-matched volunteers who participated as the T1DM controls and the T2DM controls, respectively. The total 25(OH)D levels were lowest in the 21–40 age group. The levels of both 25(OH)D3 and the total 25(OH)D were significantly higher in the T1DM and T2DM groups than in the controls (p < 0.01 in T1DM and p < 0.05 in T2DM group, respectively). The 25(OH)D2 levels were only significantly higher in T1DM patients than in the controls. The percentages of vitamin D deficiency (total 25(OH)D less than 20 ng/mL) in the T1DM, T2DM, the T1DM controls and the T2DM controls were 7.1%, 0%, 14.3% and 3.6%, respectively. The percentages of vitamin D insufficiency (total 25(OH)D less than 30 ng/mL) in the T1DM, T2DM, the T1DM controls and the T2DM controls were 26.8%, 7.3%, 54.8% and 17.9%, respectively. The percentages of vitamin D deficiency and insufficiency were significantly lower in the T1DM patients than in the T1DM controls (p < 0.01). In the present study, both type 1 and type 2 diabetes patients had higher serum 25(OH)D levels and lower percentages of vitamin D deficiency/insufficiency.     

Preliminary study on serum levels of 1,25-dihydroxyvitamin D and 25-hydroxyvitamin D in cadmium-induced renal tubular dysfunction

Serum 1,25-dihydroxyvitamin D [1,25(OH)2D] and 25-hydroxyvitamin D [25(OH)D] were measured in 7 women living in a cadmium-polluted area in Toyama, Japan. Despite the fact that these subjects had severe proximal renal tubular dysfunction showing increased fractional excretion of beta 2-microglobulin (FE beta 2-m) ranging from 9.7-49.1% with a mean of 30.7%, the levels of serum 1,25(OH)2D, which is produced in the proximal tubules, were within the normal range in 6 subjects. Significant correlations were found between 1,25(OH)2D and creatinine clearance (r = 0.802, P less than 0.05), and FE beta 2-m (r = -0.829, P less than 0.05), respectively. These results suggest that renal production of 1,25(OH)2D decreases with progression of cadmium-induced renal tubular dysfunction.     

Vitamin D and its metabolites in the pollen of pine. Part 5: Steroid hormones in the pollen of pine species

Unconjugated vitamin D and its metabolites were investigated in the pollen of Pinus nigra Ar. and Pinus sylvestris L. by TLC, HPLC and competitive radiochemical determination of 25-hydroxycholecalciferol (25-OHD3). It was found that vitamin D (D2, D3) was present in the pollen in amounts about 2 micrograms/10 g and 25-OHD3, 24,25-dihydroxycholecalciferol [24,25-(OH)2D3] and 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] between 0.1 and 3 micrograms/10 g of pollen, dependent on pollen species and methods.     

Vitamin D metabolism in Japanese quail: effects of lead exposure and dietary calcium.

Four-week-old female Japanese quail were raised either on a low calcium (0.2%) or on a normal calcium (2.3 to 3.3%) diet for 15 days and at the same time they were given drinking water containing either lead acetate (0.032, 0.32, 3.2, or 16.0 mg Pb/ml) or sodium acetate (1.75 mg acetate/ml). On the 16th day, in vitro kidney production of 1,25- and 24,25-dihydroxyvitamin D₃ [1,25-(OH)₂D₃ and 24,25-(OH)₂D₃] from tritiated 25-hydroxyvitamin D₃, plasma calcium, and phosphate, and body retention of lead (plasma, liver and femur), were determined. Under both dietary conditions, lead ingestion (0.32, 3.2 and 16.0 mg Pb/ml), when compared with sodium acetate-treated controls, resulted in the stimulation of 1,25-(OH)₂D₃ production, but 24,25-(OH)₂D₃ increased only under normal dietary calcium and higher doses (3.2 and 16.0 mg Pb/ml) of lead. Production of 24,25-(OH)₂D₃ was non-detectable in all groups given low calcium diet. Lead added in vitro to kidney homogenates of untreated, age-matched quail had no significant effect on renal hydroxylases. Lead treatment also had no significant effect on plasma calcium and phosphate concentrations. The lead concentration in plasma, liver, and femur was always higher at comparable dose levels of lead under the low calcium dietary conditions. The data indicate that subacute lead ingestion results in the stimulation of renal 25-hydroxyvitamin D₃-1-hydroxylase activity.     

Effects of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and its analogues (EB1089 and analog V) on canine adenocarcinoma (CAC-8) in nude mice

The aim of this study is to determine the effects of 1,25(OH)2D3 and its analogues on tumor growth and body weight, changes in plasma ionized calcium, parathyroid hormone-related protein (PTHrP) production, bone resorption, and the distribution of the 1,25(OH)2D3 receptor (VDR) on tumors in nude mice-bearing the canine adenocarcinoma (CAC-8). Thirty-seven nude mice were implanted subcutaneously with CAC-8. Two weeks after implantation, the mice were divided into 5 groups and injected intraperitoneally 3 times/week for 4 weeks with 5 different substrates. Group I (nontumor-bearing mice) were injected with vehicle. Groups II through V were CAC-8-bearing mice injected with the following: Grp. II, vehicle; Grp. III, analog V; Grp. IV, 1,25(OH)2D3; and Grp. V, EB1089. Our results showed that mice body weight (% change) of CAC-8-bearing mice was significantly lower than those of nontumor-bearing mice (p<0.05). CAC-8-bearing mice treated with analog V maintained their body weight better than CAC-8-bearing mice treated with either vehicle, 1,25(OH)2D3, or EB1089. A reduction of tumor growth was observed in CAC-8-bearing mice treated with 1,25(OH)2D3 and its analogues; however, the reduction was not statistically significant compared to the vehicle-treated CAC-8-bearing mice. All CAC-8-bearing mice increased osteoclastic bone resorption and hypercalcemia. Immunohistochemical staining of CAC-8 with VDR antibody demonstrated a positive reaction in nuclei of tumor cells. In conclusion, CAC-8-bearing mice treated with analog V were more active and maintained their body weight better than other CAC-8-bearing groups. Analog V-treated mice also showed no toxic side effects of hypercalcemia despite an increase in plasmaionized calcium comparable to nontumor-bearing mice. Tumor volumes of CAC-8-bearing mice treated with 1,25(OH)2D3 and its analogues were smaller than vehicle-treated CAC-8-bearing mice. This finding suggested an inhibitory effect on tumor cell growth.     

Decreased serum 24,25-dihydroxy vitamin D concentrations in children receiving chronic anticonvulsant therapy.

Serum 24,25-dihydroxy vitamin D (24,25(OH)2D) and 25-hydroxy vitamin D (25-OHD) concentrations and the ratio between the two were measured in 31 Israeli children and adolescents receiving long-term treatment with phenobarbitone or phenytoin and in controls. 24,25 (OH)2D concentrations were significantly depressed in the patients, although the 25-OHD concentrations were similar to those in the healthy controls. In four patients with radiological evidence of osteopenia very low serum 24,25(OH)2D concentrations and serum 24,25(OH)2D: 25-OHD ratios were recorded. The findings suggest that 24,25(OH)2D deficiency may play an important part in the pathogenesis of osteomalacia in patients treated with anticonvulsant drugs and provide further indirect evidence that 24,25(OH)2D is important for normal bone structure.     

Parenteral 1,25-dihydroxycholecalciferol in hepatic osteomalacia

Despite regular long-term parenteral vitamin D2 treatment, four patients with biliary cirrhosis had multiple symptoms of bone disease and bone biopsy specimens showed osteomalacia without osteoporosis. Three patients also had a proximal myopathy. Plasma calcium values (after correction for albumin), phosphorus, magnesium, and serum 25-hydroxy-vitamin D were within normal limits. Treatment with 1,25-dihydroxy-cholecalciferol (1,25-(OH)2D3) relieved symptoms in three of the four patients and improved those in the fourth. Histological examination of bone showed improvement in all four patients, but serum and urinary biochemical changes were not pronounced. We conclude that 1,25-(0H)2D3 treatment has a beneficial effect on bone and muscle in hepatic osteomalacia, either because vitamin D 1-hydroxylation fails in biliary cirrhosis or because hepatic osteomalacia is resistant to vitamin D2 metabolites.