Placental uptake and metabolism as determinants of pregnancy vitamin D status

Pregnancy 25-hydroxyvitamin D (25(OH)D) concentrations are associated with maternal and fetal health outcomes, but the underlying mechanisms have not been elucidated. Using physiological human placental perfusion approaches and intact villous explants we demonstrate a role for the placenta in regulating the relationships between maternal 25(OH)D concentrations and fetal physiology. Here, we demonstrate active placental uptake of 25(OH)D3 by endocytosis and placental metabolism of 25(OH)D3 into 24,25-dihydroxyvitamin D3 and active 1,25-dihydroxyvitamin D [1,25(OH)2D3], with subsequent release of these metabolites into both the fetal and maternal circulations. Active placental transport of 25(OH)D3 and synthesis of 1,25(OH)2D3 demonstrate that fetal supply is dependent on placental function rather than solely the availability of maternal 25(OH)D3. We demonstrate that 25(OH)D3 exposure induces rapid effects on the placental transcriptome and proteome. These map to multiple pathways central to placental function and thereby fetal development, independent of vitamin D transfer, including transcriptional activation and inflammatory responses. Our data suggest that the underlying epigenetic landscape helps dictate the transcriptional response to vitamin D treatment. This is the first quantitative study demonstrating vitamin D transfer and metabolism by the human placenta; with widespread effects on the placenta itself. These data show complex and synergistic interplay between vitamin D and the placenta, and inform possible interventions to optimise placental function to better support fetal growth and the maternal adaptations to pregnancy.

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Simple Fast Quantification of Cholecalciferol, 25-Hydroxyvitamin D and 1,25-Dihydroxyvitamin D in Adipose Tissue Using LC-HRMS/MS

Nutrients ◽

10.3390/nu11091977 ◽

2019 ◽

Vol 11 (9) ◽

pp. 1977 ◽

Cited By ~ 3

Author(s):

Laurianne Bonnet ◽

Marielle Margier ◽

Ljubica Svilar ◽

Charlene Couturier ◽

Emmanuelle Reboul ◽

...

Keyword(s):

Vitamin D ◽

Adipose Tissue ◽

Solid Phase ◽

Vitamin D Metabolites ◽

Vitamin D Metabolism ◽

Dihydroxyvitamin D3 ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

Relative Standard ◽

The Impact

Vitamin D metabolism is actively modulated in adipose tissue during obesity. To better investigate this process, we develop a specific LC-HRMS/MS method that can simultaneously quantify three vitamin D metabolites, i.e., cholecalciferol, 25-hydroxyvitamin D3 (25(OH)D3), and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) in a complex matrix, such as mouse adipose tissue and plasma. The method uses pretreatment with liquid–liquid or solid–phase extraction followed by derivatization using Amplifex® reagents to improve metabolite stability and ionization efficiency. Here, the method is optimized by co-eluting stable isotope-labelled internal standards to calibrate each analogue and to spike biological samples. Intra-day and inter-day relative standard deviations were 0.8–6.0% and 2.0–14.4%, respectively for the three derivatized metabolites. The limits of quantification (LoQ) achieved with Amplifex® derivatization were 0.02 ng/mL, 0.19 ng/mL, and 0.78 ng/mL for 1,25(OH)2D3, 25(OH)D3 and cholecalciferol, respectively. Now, for the first time, 1,25(OH)2D3 can be co-quantified with cholecalciferol and 25(OH)D3 in mouse adipose tissue. This validated method is successfully applied to study the impact of obesity on vitamin D status in mice.

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Reshaping the way we view vitamin D signalling and the role of vitamin D in health

Nutrition Research Reviews ◽

10.1079/nrr200480 ◽

2004 ◽

Vol 17 (2) ◽

pp. 241-248 ◽

Cited By ~ 4

Author(s):

James C. Fleet ◽

Jie Hong ◽

Zhentao Zhang

Keyword(s):

Vitamin D ◽

Transcriptional Activation ◽

Mode Of Action ◽

Uv Light ◽

Active Form ◽

Vitamin D Metabolism ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

25 Hydroxyvitamin D ◽

Molecular Mode

AbstractAlthough the biological requirement for vitamin D can be met by epidermal exposure to UV light, there are a number of conditions where this production does not occur or is not sufficient to meet biological needs. When this happens, vitamin D must be consumed and is a nutrient. However, two distinct observations have caused researchers to rethink certain dogma in vitamin D biology. First, it appears that in addition to the hormonally active form of 1,25 dihydroxyvitamin D (1,25(OH)2D), circulating levels of 25 hydroxyvitamin D have a critical importance for optimal human health. This and other data suggest that extra-renal production of 1,25(OH)2D contributes to Ca homeostasis and cancer prevention. Second, in addition to its role in the transcriptional activation of genes through the vitamin D receptor there is now compelling evidence that 1,25(OH)2D has a second molecular mode of action; the rapid activation of second-messenger and kinase pathways. The purpose of this second mode of action is only now being explored. The present review will discuss how these two areas are reshaping our understanding of vitamin D metabolism and action.

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Inhibitory effect of NF-κB on 1,25-dihydroxyvitamin D3 and retinoid X receptor function

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.2000.279.1.e213 ◽

2000 ◽

Vol 279 (1) ◽

pp. E213-E220 ◽

Cited By ~ 27

Author(s):

Paul K. Farmer ◽

Xiaofei He ◽

M. Lienhard Schmitz ◽

Janet Rubin ◽

Mark S. Nanes

Keyword(s):

Vitamin D ◽

Dna Sequences ◽

Transcriptional Activation ◽

Inhibitory Effect ◽

Retinoid X Receptor ◽

Mobility Shift ◽

Dihydroxyvitamin D3 ◽

Dihydroxyvitamin D ◽

Tnf Α ◽

Factor Α

Responsiveness to 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] may be diminished in osteoporosis and inflammatory arthritis. The inflammatory cytokine tumor necrosis factor-α (TNF-α) is produced in excess in these disorders and has been shown to decrease osteoblast transcriptional responsiveness to vitamin D and to inhibit the binding of the vitamin D receptor (VDR) and its nuclear partner the retinoid X receptor (RXR) to DNA. Previous studies have shown that a vitamin D (VDRE) or retinoid X DNA response element (RXRE) is sufficient to confer TNF-α inhibition of vitamin D or retinoid-stimulated transcription in the absence of known TNF-α-responsive DNA sequences. We tested the hypothesis that the TNF-α-stimulated transcription factor nuclear factor (NF)-κB could, in part, mediate TNF-α action by inhibiting the transcriptional potency of the VDR and RXR at their cognate cis regulatory sites. Osteoblastic ROS 17/2.8 cells transfected with a dose of NF-κB comparable to that stimulated by TNF-α decreased 1,25(OH)2D3-stimulated transcription. This inhibitory effect of NF-κB was not observed on basal transcription of a heterologous reporter in the absence of the VDRE. The effects of NF-κB and TNF-α were comparable but not additive. COS-7 cells were cotransfected with reporters under the regulation of VDRE or RXRE along with vectors expressing VDR, RXR, and NF-κB nuclear proteins. Reconstituted NF-κB and the NF-κB subunit p65 alone, but not p50, dose dependently suppressed basal and ligand-stimulated transcription. p65 overexpression completely abrogated enhanced VDRE-mediated transcriptional activity in response to 1,25(OH)2D3. Electrophoretic mobility shift experiments did not reveal a direct effect of recombinant NF-κB or its individual subunits on the binding of heterodimeric VDR-RXR to DNA. These results suggest that TNF-α inhibition of hormone-stimulated transcriptional activation may be mediated by activation of NF-κB. In contrast, the inhibitory effect of TNF-α on binding of receptors to DNA is unlikely to be mediated by NF-κB and is not necessary for inhibition of transcription.

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1,25-Dihydroxyvitamin D3 downregulates the rat intestinal vitamin D3-25-hydroxylaseCYP27A

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.2001.281.2.e315 ◽

2001 ◽

Vol 281 (2) ◽

pp. E315-E325 ◽

Cited By ~ 22

Author(s):

Catherine Theodoropoulos ◽

Christian Demers ◽

Ali Mirshahi ◽

Marielle Gascon-Barré

Keyword(s):

Vitamin D ◽

Direct Action ◽

Dihydroxyvitamin D3 ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

Protein Levels ◽

25 Hydroxyvitamin D ◽

Extrahepatic Tissues ◽

Mrna Half Life

The vitamin D3-25-hydroxylase CYP27A is located predominantly in liver, but its expression is also detected in extrahepatic tissues. Our aim was to evaluate the regulation of CYP27A by vitamin D3 (D3) or its metabolites in rat duodena. Vitamin D-depleted rats were repleted with D3, 25-hydroxyvitamin D (25OHD), or 1,25-dihydroxyvitamin D3[1,25(OH)2D3] or acutely injected 1,25(OH)2D3 to investigate the mechanisms of action of the hormone. All D3 compounds led to a progressive decrease in CYP27A mRNA, with levels after D3 representing 20% of that observed in D depletion. 25OHD decreased CYP27A mRNA by 55%, whereas 1,25(OH)2D3 led to a 40% decrease, which was accompanied by a 31% decrease in CYP27A protein levels and an 89% decrease in enzyme activity. Peak circulating 1,25(OH)2D3 concentrations were, however, the highest in D3-repleted, followed by 25OHD- and 1,25(OH)2D3-repleted animals. 1,25(OH)2D3 resulted in a decrease in both CYP27A mRNA half-life and transcription rate. Our data illustrate that the intestine expresses the D3-25-hydroxylase and that the gene is highly regulated in vivo through a direct action of 1,25(OH)2D3 or through the local production of D3 metabolites.

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Vitamin D attenuates inflammation in CFTR knockdown intestinal epithelial cells but has no effect in cells with intact CFTR

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00060.2015 ◽

2016 ◽

Vol 310 (8) ◽

pp. G539-G549 ◽

Cited By ~ 6

Author(s):

Geneviève Morin ◽

Valérie Orlando ◽

Karoline St-Martin Crites ◽

Natacha Patey ◽

Geneviève Mailhot

Keyword(s):

Vitamin D ◽

Intestinal Inflammation ◽

Inflammatory Responses ◽

Vitamin D Metabolites ◽

Intestinal Epithelial ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

1,25 Dihydroxyvitamin D ◽

25 Hydroxyvitamin D ◽

Anti Inflammatory

The cystic fibrosis (CF) intestine is characterized by chronic inflammation. CF patients are instructed to ingest supplemental vitamin D on a daily basis thereby exposing their intestinal tract to pharmacological amounts of this vitamin. It has been shown that vitamin D exerts intestinal anti-inflammatory properties. We therefore postulate that vitamin D may be beneficial in the management of CF intestinal inflammation by attenuating cellular inflammatory responses. In this study, we investigated the anti-inflammatory effects of the oral form of vitamin D3 (cholecalciferol) and its metabolites, 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3, on cytokine-induced inflammatory responses in intestinal epithelial Caco-2/15 cells with intact expression of CF transmembrane conductance regulator (CFTR) and knockdown for CFTR. We show that 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 inhibited p38MAPK phosphorylation and that these effects were not mediated by changes in the expression of MAPK phosphatase-1 (MKP-1). However, 1,25-dihydroxyvitamin D3 exhibited superior anti-inflammatory effects as it furthermore reduced cytokine-induced NF-κB nuclear translocation and interleukin-8 mRNA stability and secretion. Intriguingly, the anti-inflammatory effects of vitamin D metabolites were only observed in CFTR knockdown cells, which may be explained by alterations in its catabolism associated with changes in CYP24A1 expression. These observations were supported in vivo whereby Cftr −/− mice fed large amounts of vitamin D3 for 2 mo led to a reduction in the number of eosinophils and apoptotic cells in the duodenal mucosa of females but not males. Altogether, these findings suggest that vitamin D exerts intestinal anti-inflammatory actions under specific circumstances and may thus prove beneficial in CF.

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Changes in serum levels of 1,25-dihydroxyvitamin D3, calcium and phosphorus with age and vitamin D status in chickens

British Journal Of Nutrition ◽

10.1079/bjn19840099 ◽

1984 ◽

Vol 52 (2) ◽

pp. 329-334 ◽

Cited By ~ 7

Author(s):

Saleh H. Sedrani

Keyword(s):

Vitamin D ◽

Serum Level ◽

Sexual Maturity ◽

Serum Levels ◽

Minimal Amount ◽

Dihydroxyvitamin D3 ◽

Dietary Regimen ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

25 Hydroxyvitamin D

1. The effects of vitamin D3(D3) on serum levels of 1, 25-dihydroxyvitamin D3(1, 25(OH)2D3), ionic calcium, total Ca and phosphorus in chicks were studied from the time of hatching until sexual maturity.2. Chicks fed on a diet low in D3showed a serum level of 1, 25(OH)2D3higher than that in chicks on a normal-D3diet, for both sexes and at any given age.3. A dramatic increase in the serum level of 1, 25(OH)2D3occurred in female birds approaching sexual maturity and in laying hens raised on the low-D3diet the level was five times that of their counterparts raised on a normal-D3diet.4. Theserum 1, 25(OH)2D3levelin adultmalesin thelow-D3groups wasseven timesthatofthoseon thenormal-D3diet.5. The serum level of 25-hydroxyvitamin D3remained relatively unchanged at weeks 2 and 15 in birds on a low D3intake as well as in those fed on a normal-D3diet. Nevertheless, the levels of 25-hydroxyvitamin D3were different between the two groups.6. No significant change was observed in the level of ionized serum Ca in relation to dietary regimen, but there was an increase in total Ca concentration in females with the onset of reproduction.7. The serum P level decreased gradually with age, reaching a minimum value 3 and 8 weeks before laying commenced in the groups on low- and normal-D3diets respectively. An increase was observed when the hens began laying.8. Chicks adapted to a low-D3diet by elevation of their plasma level of 1, 25(OH)2D3. The mechanism by which this is achieved is not known, but the results suggest that parathyroid hormone, Ca and P are unlikely to play roles in the adaptive increase in the level of 1, 25(OH)2D3in the blood of chicks given a minimal amount of D3. The possibility that the rate of degradation of 1, 25(OH)2D3is greatly reduced under these conditions cannot be excluded and this could account for the level of this metabolite in those birds.

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Functional Cooperation between CCAAT/Enhancer-Binding Proteins and the Vitamin D Receptor in Regulation of 25-Hydroxyvitamin D3 24-Hydroxylase

Molecular and Cellular Biology ◽

10.1128/mcb.25.1.472-487.2005 ◽

2005 ◽

Vol 25 (1) ◽

pp. 472-487 ◽

Cited By ~ 80

Author(s):

Puneet Dhawan ◽

Xiaorong Peng ◽

Amelia L. M. Sutton ◽

Paul N. MacDonald ◽

Colleen M. Croniger ◽

...

Keyword(s):

Vitamin D ◽

Vitamin D Receptor ◽

Cross Talk ◽

Transcriptional Response ◽

Dominant Negative ◽

Osteoblastic Cells ◽

Cooperative Effects ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

25 Hydroxyvitamin D

ABSTRACT 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] induces the synthesis of 25-hydroxyvitamin D3 24-hydroxylase [24(OH)ase], an enzyme involved in its catabolism, thereby regulating its own metabolism. Here we demonstrate that CCAAT enhancer binding protein β (C/EBPβ) is induced by 1,25(OH)2D3 in kidney and in osteoblastic cells and is a potent enhancer of vitamin D receptor (VDR)-mediated 24(OH)ase transcription. Transfection studies indicate that 1,25(OH)2D3 induction of 24(OH)ase transcription is enhanced a maximum of 10-fold by C/EBPβ. Suppression of 1,25(OH)2D3-induced 24(OH)ase transcription was observed with dominant negative C/EBP or osteoblastic cells from C/EBPβ−/− mice. A C/EBP site was identified at positions −395 to −388 (−395/−388) in the rat 24(OH)ase promoter. Mutation of this site inhibited C/EBPβ binding and markedly attenuated the transcriptional response to C/EBPβ. We also report the cooperation of CBP/p300 with C/EBPβ in regulating VDR-mediated 24(OH)ase transcription. We found that not only 1,25(OH)2D3 but also parathyroid hormone (PTH) can induce C/EBPβ expression in osteoblastic cells. PTH potentiated the induction of C/EBPβ and 24(OH)ase expression in response to 1,25(OH)2D3 in osteoblastic cells. Data with the human VDR promoter (which contains two putative C/EBP sites) indicate a role for C/EBPβ in the protein kinase A-mediated induction of VDR transcription. From this study a fundamental role has been established for the first time for cooperative effects and cross talk between the C/EBP family of transcription factors and VDR in 1,25(OH)2D3-induced transcription. These findings also indicate a novel role for C/EBPβ in the cross talk between PTH and 1,25(OH)2D3 that involves the regulation of VDR transcription.

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Activation of Receptor Activator of NF-κB Ligand Gene Expression by 1,25-Dihydroxyvitamin D3 Is Mediated through Multiple Long-Range Enhancers

Molecular and Cellular Biology ◽

10.1128/mcb.00353-06 ◽

2006 ◽

Vol 26 (17) ◽

pp. 6469-6486 ◽

Cited By ~ 166

Author(s):

Sungtae Kim ◽

Miwa Yamazaki ◽

Lee A. Zella ◽

Nirupama K. Shevde ◽

J. Wesley Pike

Keyword(s):

Gene Expression ◽

Vitamin D ◽

Rna Polymerase Ii ◽

Transcriptional Activation ◽

Molecular Mechanisms ◽

Chromatin Modification ◽

Response Element ◽

Dihydroxyvitamin D3 ◽

Specific Element ◽

Dihydroxyvitamin D

ABSTRACT RANKL is a tumor necrosis factor (TNF)-like factor secreted by mesenchymal cells, osteoblast derivatives, and T cells that is essential for osteoclastogenesis. In osteoblasts, RANKL expression is regulated by two major calcemic hormones, 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and parathyroid hormone (PTH), as well as by several inflammatory/osteoclastogenic cytokines; the molecular mechanisms for this regulation are unclear. To identify such mechanisms, we screened a DNA microarray which tiled across the entire mouse RankL gene locus at a 50-bp resolution using chromatin immunoprecipitation (ChIP)-derived DNA precipitated with antibodies to the vitamin D receptor (VDR) and the retinoid X receptor (RXR). Five sites of dimer interaction were observed on the RankL gene centered at 16, 22, 60, 69, and 76 kb upstream of the TSS. These regions contained binding sites for not only VDR and RXR, but also the glucocorticoid receptor (GR). The most distant of these regions, termed the distal control region (RL-DCR), conferred both VDR-dependent 1,25(OH)2D3 and GR-dependent glucocorticoid (GC) responses. We mapped these activities to an unusual but functionally active vitamin D response element and to several potential GC response elements located over a more extensive region within the RL-DCR. An evolutionarily conserved region within the human RANKL gene contained a similar vitamin D response element and exhibited an equivalent behavior. Importantly, hormonal activation of the RankL gene was also associated with chromatin modification and RNA polymerase II recruitment. Our studies demonstrate that regulation of RankL gene expression by 1,25(OH)2D3 is complex and mediated by at least five distal regions, one of which contains a specific element capable of mediating direct transcriptional activation.

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Serum 25-hydroxyvitamin D3 and 24R,25-dihydroxyvitamin D3 concentrations in adult dogs are more substantially increased by oral supplementation of 25-hydroxyvitamin D3 than by vitamin D3

Journal of Nutritional Science ◽

10.1017/jns.2017.8 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 2

Author(s):

Lauren R. Young ◽

Robert C. Backus

Keyword(s):

Vitamin D ◽

Hplc Method ◽

Vitamin D Status ◽

Dihydroxyvitamin D3 ◽

Oral Supplementation ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

Supplementation Period ◽

Single Cross ◽

25 Hydroxyvitamin D

AbstractWe previously found a weak response in serum 25-hydroxyvitamin D3 (25(OH)D3) concentrations when dogs were supplemented with oral vitamin D3 (D3). In the present study, we determined the relative potency of oral 25(OH)D3 compared with D3 for increasing vitamin D status in dogs with low serum 25(OH)D concentrations. Four male and three female, 4-year-old, intact, lean, genetically related, Chinese-crested/beagle dogs were studied in a randomised, single cross-over trial. After feeding a low-vitamin D diet (<4 IU/100 g) for 30 d, four dogs received daily D3 supplementation at 2·3 µg/kg body weight0·75, while three dogs received a molar equivalency as 25(OH)D3. The supplements, dissolved in ethanol, were applied to a commercial treat for consumption. Serum 25(OH)D3 and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) were analysed weekly using a validated HPLC method. Both supplementations increased (P ≤ 0·01) serum 25(OH)D3 concentrations. However, oral 25(OH)D3 resulted in greater (P < 0·0001) concentrations than D3 by week 1, with a difference of 173 % (P < 0·0001) by week 2. The supplementation period was limited to 14 d after serum 25(OH)D3 concentrations were not appearing to plateau. Thereafter, a washout period of 1 month separated the cross-over. Following 25(OH)D3, but not D3 supplementation, serum 24R,25(OH)2D3 concentrations increased (P ≤ 0·02), 3 to 5 weeks after initiating supplementation. Vitamin D status, as indicated by serum 25(OH)D3 and 24R,25(OH)2D3 concentrations, is more rapidly and efficiently increased in adult dogs by oral supplementation of 25(OH)D3 than D3.

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Vitamin D upregulates the macrophage complement receptor immunoglobulin in innate immunity to microbial pathogens

Communications Biology ◽

10.1038/s42003-021-01943-3 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Annabelle G. Small ◽

Sarah Harvey ◽

Jaspreet Kaur ◽

Trishni Putty ◽

Alex Quach ◽

...

Keyword(s):

Vitamin D ◽

Innate Immunity ◽

Surface Expression ◽

Microbial Pathogens ◽

Cell Surface Expression ◽

Complement Receptor ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

Multistep Process ◽

25 Hydroxyvitamin D

AbstractVitamin D deficiency remains a global concern. This ‘sunshine’ vitamin is converted through a multistep process to active 1,25-dihydroxyvitamin D3 (1,25D), the final step of which can occur in macrophages. Here we demonstrate a role for vitamin D in innate immunity. The expression of the complement receptor immunoglobulin (CRIg), which plays an important role in innate immunity, is upregulated by 1,25D in human macrophages. Monocytes cultured in 1,25D differentiated into macrophages displaying increased CRIg mRNA, protein and cell surface expression but not in classical complement receptors, CR3 and CR4. This was associated with increases in phagocytosis of complement opsonised Staphylococcus aureus and Candida albicans. Treating macrophages with 1,25D for 24 h also increases CRIg expression. While treating macrophages with 25-hydroxyvitamin D3 does not increase CRIg expression, added together with the toll like receptor 2 agonist, triacylated lipopeptide, Pam3CSK4, which promotes the conversion of 25-hydroxyvitamin D3 to 1,25D, leads to an increase in CRIg expression and increases in CYP27B1 mRNA. These findings suggest that macrophages harbour a vitamin D-primed innate defence mechanism, involving CRIg.

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