Effects Of Vitamin D3 Metabolites Research Articles

Inhibition of hepatitis C virus by vitamin D.

Until the development of direct-acting antivirals (DAAs), interferon (IFN)-based therapy had been the primary treatment strategy for patients with chronic hepatitis C, even though this therapy has a therapeutic limitations and considerable side effects. Therefore, many efforts have been made to improve the efficacy of treatment. Several clinical studies have clearly shown that supplementation with vitamin D of IFN-based therapy improves treatment efficacy. To clarify the molecular mechanisms of the effect of vitamin D on IFN-based therapy, several researchers have performed basic research with cell culture models of hepatitis C virus (HCV). Consequently, two vitamin D3 metabolites, 25-hydroxyvitamin D3 (25-(OH)D3) and 1α,25-dihydroxyvitamin D3 (1α,25-(OH)2D3), have been suggested to have anti-HCV effects. 25-(OH)D3 inhibits HCV production by suppressing infectious virus assembly through reducing apolipoprotein expression, while 1α,25-(OH)2D3 inhibits HCV production by modulating IFN signaling and/or inducing various host factors associated with the inhibition of viral genome replication. In addition, an antimicrobial peptide, LL-37, which is known to be partly regulated by vitamin D, was also reported to exhibit an anti-HCV effect by disrupting infectious viral particles directly. In conclusion, vitamin D3 supplementation improves the response rate of IFN-based therapy via the direct and/or indirect anti-HCV effects of vitamin D3 metabolites.

Effect of different vitamin D3 metabolites on intestinal calcium homeostasis-related gene expression in broiler chickens

ABSTRACT The purpose of this study was to investigate the effects of vitamin D3 metabolites 1α-hydroxycholecalciferol (1α(OH)D3), 25-hydroxycholecalciferol (25(OH)2D3), and 1,25-dihydroxycholecalciferol (1,25(OH)2D3) on growth performance, bone quality, and intestinal calcium homeostasis-related gene expression in broiler chickens. One-day-old broilers were fed a basal diet and basal diet containing different vitamin D3 metabolites. The body weight, feed intake, and feed conversion ratio in control and experimental broilers were measured to assess the growth performance, mineral levels, and bone breaking strength. The duodenum [...]

Vitamin D3 Metabolism in Human Glioblastoma Multiforme: Functionality of CYP27B1 Splice Variants, Metabolism of Calcidiol, and Effect of Calcitriol

A better understanding of the vitamin D(3) metabolism is required to evaluate its potential therapeutic value for cancers. Here, we set out to contribute to the understanding of vitamin D(3) metabolism in glioblastoma multiforme. We did nested touchdown reverse transcription-PCR (RT-PCR) to identify CYP27B1 splice variants and real-time RT-PCR to quantify the expression of CYP27B1. A cell line was treated with calcitriol to determine the effect on the expression of CYP27B1, 1alpha,25-dihydroxyvitamin D(3)-24-hydroxylase (CYP24), and vitamin D(3) receptor (VDR). We generated three antibodies for the specific detection of CYP27B1 and splice variants. High-performance TLC was done to determine the endogenous CYP27B1 activity and the functionality of CYP27B1 splice variants. Using WST-1 assay, we determined the effect of vitamin D(3) metabolites on proliferation. We report a total of 16 splice variants of CYP27B1 in glioblastoma multiforme and a different expression of CYP27B1 and variants between glioblastoma multiforme and normal tissues. We found preliminary evidence for enzymatic activity of endogenous CYP27B1 in glioblastoma multiforme cell cultures but not for the functionality of the splice variants. By adding calcitriol, we found a proliferative effect for some cell lines depending on the dose of calcitriol. The administration of calcitriol led to an elevated expression of CYP27B1 and CYP24 but left the expression of the VDR unaltered. Our findings show that glioblastoma multiforme cell lines metabolize calcidiol. In addition, we show various effects mediated by calcitriol. We found a special vitamin D(3) metabolism and mode of action in glioblastoma multiforme that has to be taken into account in future vitamin D(3)-related therapies.

The synergistic effect of TGF beta and 24,25-(OH)2D3 on resting zone chondrocytes is metabolite-specific and mediated by PKC.

Transforming growth factor-beta (TGF beta) and 24,25-(OH)2D3 play a major role in chondrocyte maturation during endochondral bone formation. TGF beta and vitamin D metabolites when added separately to resting zone (RC) or growth zone (GC) chondrocyte cultures, activate protein kinase C (PKC). The present study determined whether there is an additive or synergistic effect of vitamin D3 metabolites and TGF beta on alkaline phosphatase and PKC specific activities and whether this effect is cell maturation-dependent. GC and RC chondrocytes were isolated from rat costochondral cartilage and cultured to fourth passage. The cells were incubated with vitamin D3 metabolites and TGF beta alone or in combination, and the specific activity of alkaline phosphatase, as well as the specific activity of PKC, were measured. The addition of 24,25-(OH)2D3 with TGF beta to RC cells caused a synergistic effect on alkaline phosphatase activity; this result was not found if the vitamin D3 metabolite was replaced by 1,25-(OH)2D3. The addition of 1,25-(OH)2D3 or 24,25-(OH)2D3 with TGF beta on GC cells had no synergistic or additive effect. The addition of 24,25-(OH)2D3 and TGF beta for 12 hours caused a synergistic effect on PKC activity; this effect was also observed if TGF beta was added first for 12 hours and 24,25-(OH)2D3 for the last 90 min. However, the addition of 24,25-(OH)2D3 for 90 min followed by the addition of TGF beta for an additional 10.5 hours had no synergistic effect. This study indicates that TGF beta and 24,25-(OH)2D3 have a synergistic effect on chondrocyte differentiation as well as on PKC activity, suggesting that the synergistic effect of 24,25-(OH)2D3 and TGF beta on chondrocyte differentiation may be mediated through activation of PKC. The synergistic effect of 24,25-(OH)2D3 and TGF beta was cell maturation dependent.

Effects of vitamin D3metabolites on thyrotropin secretion from rat pituitary cells in culture

The effects of several vitamin D3 metabolites on TSH secretion were investigated using rat pituitary cells in culture. After a 24-h incubation in Dulbecco's medium, only 25,26-dihydroxyvitamin D3 [25,26(OH)2D3] at 10(-6) M increased the basal TSH secretion (36% above control). The TRH-induced TSH secretion increased dose-dependently with increasing concentrations of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] showing a significant increase at 10(-8) M (41% above control). At 10(-5) M 25,26(OH)2D3 the TRH-stimulated TSH secretion was also increased (63% above control). In medium containing charcoal-treated serum, 1, 24,25-trihydroxyvitamin D3 [1,24,25(OH)3D3] decreased the TRH-induced TSH secretion dose dependently (34% below control at 10(-8) M), while the effects of both 1,25(OH)2D3 and 25,26(OH)2D3 were blunted. Our results indicate that several vitamin D3 metabolites may affect pituitary TSH secretion.

Effects of vitamin D3 metabolites in ovariectomized rats.

The effects of two vitamin D3 metabolites, 24R,25-dihydroxyvitamin D3 and 1 alpha,25-dihydroxyvitamin D3, were investigated in ovariectomized rats. The amount of ash in the femur on a defatted dry weight basis was significantly greater in rats treated with 1 microgram/kg 24R,25-dihydroxyvitamin D3, or 0.01 or 0.1 microgram/kg 1 alpha,25-dihydroxyvitamin D3 than in the controls. The concentration of bone gla protein in serum and amounts in the femur were significantly greater in rats treated with 1 or 10 micrograms/kg 24R,25-dihydroxyvitamin D3, but not those given 1 alpha,25-dihydroxyvitamin D3 compared with the controls. These results suggest that 24R,25-dihydroxyvitamin D3 increased bone mass probably through the stimulation of bone formation.

Effect of vitamin D3 metabolites on calcium and phosphorus metabolism in chick embryos.

The biochemical nature of the physiological defect found in chick embryos from hens supported on 1,25-dihydroxyvitamin D3 as their sole source of vitamin D is described. Vitamin D-deficient hens (44-wk-old) were divided into six groups of five and dosed daily for 19 wk with either 2.0 micrograms of 25-hydroxyvitamin D3, 2.0 micrograms of 24,24-difluoro-25-hydroxy-vitamin D3, 0.4 micrograms of 1,25-dihydroxyvitamin D3, 2.0 micrograms of 24,25-dihydroxyvitamin D3, 0.4 micrograms of 1,25-dihydroxyvitamin D3 plus 2.0 micrograms of 24,25-dihydroxyvitamin D3, or vehicle only. Normal embryonic development was found in eggs from hens given 25-hydroxyvitamin D3 or 24,24-difluoro-25-hydroxyvitamin D3, whereas embryos from hens given 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D3, or their combination were abnormal and failed to hatch. Embryos from hens fed 1,25-dihydroxyvitamin D3 and/or 24,25-dihydroxyvitamin D3 had vitamin D deficiency: low bone ash, low plasma calcium, low total body calcium, and extremely high plasma phosphorus. Because the shell is the major source of calcium for the developing embryo, calcium transport from the shell to the embryos across the chorioallantoic membrane apparently fails, giving rise to the observed defects in embryonic development.

Effects of calcium and (inorganic) phosphate on bone resorption produced by vitamin D3 metabolites in vitro.

The effects of changing the concentrations of calcium (Ca2+) and inorganic phosphate (Pi) on 45Ca release from prelabeled fetal rat bones caused by parathyroid hormone (PTH), 25-hydroxyvitamin D3 (25-OH-D3) and 1, 25-dihydroxyvitamin D3 (1,25-(OH)2-D3) were tested, in vitro. Increasing total (Ca2+) from 1.25 mM to 2.5 mM caused a significant increase in bone resorption caused by submaximal concentrations of these materials. Further increase in total (Ca2+) to 3.5 mM abolished this potentiating effect of bone resorption. Increasing the (Pi) from 1 mM to 2-3 mM caused significant inhibition of bone resorption caused by these resorbing materials. In addition, a (Pi) of 3 mM caused significant inhibition of higher concentrations of PTH and 25-OH-D3. (Pi) of 2 and 3 mM also caused significant inhibition of 45Ca release from control bones. It is concluded that a moderate increase in medium (Ca2+) mimics the effect of these bone resorbing materials on bone. Phosphate appears to be a direct inhibitor of bone resorption. Therefore, changing the (Ca2+) and (Pi) in the medium can affect the bone resorbing effect of vitamin D3 metabolites, similar to their effect on PTH.

Effects of vitamin D3 metabolites on calcium fluxes in intact chicken skeletal muscle and myoblasts culturedin vitro

25-hydroxycholecalciferol (25OHD3) and 1,25-dihydroxycholecalciferol (1,25(OH)2D3) at physiological concentrations exerted direct effects on Ca fluxes in cultured vitamin D-deficient chick soleus muscle and myoblasts. Isotopic desaturation curves of soleus muscle prelabeled with 45Ca indicated that the action of 25OHD3 is localized in a slow-exchangeable Ca pool where it stimulates net Ca uptake. On the other hand, the predominant effects of 1,25(OH)2D3 consist in an increase of the rate constant of Ca efflux of this pool and in an increase of net Ca uptake in a fast-exchangeable pool. 24,25-dihydroxycholecalciferal proved to be inactive on both Ca uptake and efflux. In addition, 1,25(OH)2D3 significantly increased 45Ca labeling of cultured chick myoblasts. These effects were accompanied by changes in the growth and differentiation of the cultures. The results suggest a direct involvement in vivo of 25OHD3 and 1,25(OH)2D3 on muscle cellular calcium.

Vitamin D3 metabolites and PTH synergistically stimulate bone formation of chick embryonic femur in vitro.

Bone remodelling depends on a balance formation and resorption. Little is known of the biological factors involved in bone formation, whereas there is much evidence that physiological factors, such as parathyroid hormone (PTH) and active metabolites of vitamin D3, influence resorption. Cells responsible for osteogenesis and and osteoclasis occur in close proximity, suggesting that both might be controlled by the same physiological factors in the micro-environment of the body fluid. To clarify the factors stimulating bone formation or accelerating osteogenic cell activity, we have examined the effects of vitamin D3 metabolites and PTH on bone formation in vitro. We report here that combinations of those metabolites especially 1 alpha,25-(OH)2D3 and 24R,25-(OH)2D3, directly stimulate calcification on bone synergistically with PTH.

Interaction of 24,25-dihydroxyvitamin D3 and parathyroid hormone on bone enzymes in vitro.

The in vitro effects of vitamin D3 metabolites, parathyroid extract (PTE), purified parathyroid hormone (bPTH), vitamin A, and heparin on acid and alkaline phosphatases in rat or mouse calvaria in culture were investigated. Results show that: (a) when compared to values found in half calvaria incubated for 24 h in control medium, the bone acid and alkaline phosphatase content is significantly higher in paired halves incubated with PTE (L USP/ml), bPTH (4 x 10(-8)M), heparin (5 USP/ml), vitamin A (23 USP/ml), 25-(OH)D3 (2.5 x 10(-11) to 2.5 x 10(-8)M), 24,25-(OH)2D3, and 1,25-(OH)2D3 (2.5 x 10(-12) to 2.5 x 10(-7M); (b) the presence of 24,25-(OH)2D3 at low concentrations in the incubation medium decreases significantly the PTE, bPTH, vitamin A, or heparin induced stimulation of the phosphatase activities. This interaction is also observed when measuring beta glucuronidase and glucose-6-phosphatase activities and 45Ca release from previously labeled mouse calvaria; (c) a similar activity could not be found with 1,25-(OH)2D3 suggesting that 24,25-(OH)2D3 may have a specific role in bone metabolism.