ABSTRACT
BACKGROUND AND OBJECTIVE: Vitamin D-1,25(OH)2 D3 or 1,25D3-maintains healthy osseous tissue, stimulates the production of the antimicrobial peptide cathelicidin and has anti-inflammatory effects, but it can cause hypercalcemia. Evidence links diminished serum levels of 1,25D3 with increased gingival inflammation. Periodontitis progression is associated with increased local production of inflammatory mediators by immune cells and gingival fibroblasts. These include interleukin (IL)-6, a regulator of osteoclastic bone resorption, and the neutrophil chemoattractant IL-8, both regulated by signaling pathways, including NF-κB and MAPK/AP-1. The objectives were to determine the effects of 1,25D3 or a non-calcemic analog, 20-hydroxyvitamin D3 -20(OH)D3 or 20D3-on IL-1ß-stimulated IL-6 and IL-8 production, and NF-κB and MAPK/AP-1 activation, by human gingival fibroblasts. MATERIAL AND METHODS: Human gingival fibroblasts were incubated ± IL-1ß, with or without exposure to 1,25D3 or 20D3. IL-6 and IL-8 in culture supernatants were measured by enzyme-linked immunosorbent assay. NF-κB (p65) and AP-1 (phospho-cJun) and were measured in nuclear extracts via binding to specific oligonucleotides. Data were analyzed using ANOVA and Scheffe's F procedure for post hoc comparisons. RESULTS: IL-1ß-stimulated IL-6 and IL-8 levels were both significantly inhibited (40%-60%) (P<.045) by 1,25D3, but not 20D3 (0%-15% inhibition, not statistically significant). Both 1,25D3 and 20D3 significantly and similarly inhibited IL-1ß-stimulated nuclear levels of p65 and phospho-cJun (P<.02). CONCLUSION: Reduction of the activation of NF-κB and AP-1 alone is not able to inhibit strongly the IL-1ß stimulated IL-6 and IL-8 gene expression. 1,25D3 but not 20D3 may affect some of the many other factors/processes/pathways that in turn regulate the expression of these genes. However, the results suggest that topical application of ligands of the vitamin D receptor may be useful in the local treatment of periodontitis while reducing adverse systemic effects.
Subject(s)
Calcifediol/analogs & derivatives , Calcitriol/antagonists & inhibitors , Fibroblasts/drug effects , Gingiva/drug effects , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Interleukin-8/metabolism , Calcifediol/antagonists & inhibitors , Cell Line , Cell Survival/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Gene Expression Regulation , Gingiva/metabolism , Humans , Interleukin-6/genetics , Interleukin-8/genetics , NF-kappa B/drug effects , Periodontitis/metabolism , Signal Transduction/drug effects , Transcription Factor AP-1/drug effectsABSTRACT
It is proved that at experimental diabetes the calcium content in hepatocytes is disturbed. This disorder is mostly shown in increase of calcium content in hepatocytes and in decrease of accumulation of these ions in mitochondrias. One of the possible reasons of changes at this pathology is the change of lipid content of hepatocytes, mitochondrias and microsomes. It is proved that the in vitro model systems Ca ions inhibits the synthesis of 25-hydroxyvitamin D3 by hepatocytes.
Subject(s)
Biological Transport, Active/drug effects , Calcifediol/antagonists & inhibitors , Calcium/metabolism , Diabetes Mellitus, Experimental/metabolism , Hepatocytes/metabolism , Membrane Lipids/analysis , Animals , Calcifediol/biosynthesis , Calcium/pharmacology , Cell Membrane/chemistry , Hepatocytes/chemistry , Ion Transport/drug effects , Microsomes/metabolism , Mitochondria, Liver/metabolism , Rats , Rats, WistarABSTRACT
To enzymatically synthesize vitamin D derivatives, we screened about 300 Streptomyces sp. strains. Streptomyces sclerotialus FERM BP-1370 and Streptomyces roseoporus FERM BP-1574 were found to have the ability to convert 25-hydroxyvitamin D3 and 1 alpha-hydroxyvitamin D3, respectively, to 1 alpha, 25-dihydroxyvitamin D3. The average rates of 1 alpha hydroxylation of 25-hydroxyvitamin D3 were 6.9 micrograms liter-1 min-1 with FERM BP-1370 and 7.0 micrograms liter-1 min-1 with FERM BP-1574. The specific cytochrome P-450 inhibitors carbon monoxide, SKF-525-A, and metyrapone inhibited the hydroxylation of 1 alpha- and 25-hydroxyvitamin D3 to 1 alpha, 25-dihydroxyvitamin D3 by FERM BP-1370 and FERM BP-1574. The cytochromes P-450 of these strains were detected by reduced CO difference spectra in the whole-cell suspensions. The appearance of cytochrome P-450 suggests that the cytochromes P-450 of FERM BP-1370 and FERM BP-1574 carry out the hydroxylation of 25- and 1 alpha-hydroxyvitamin D3 to 1 alpha, 25-dihydroxyvitamin D3.
Subject(s)
Calcifediol/metabolism , Calcitriol/metabolism , Hydroxycholecalciferols/metabolism , Streptomyces/metabolism , Biotransformation , Calcifediol/antagonists & inhibitors , Calcifediol/pharmacology , Calcitriol/antagonists & inhibitors , Calcitriol/pharmacology , Cytochrome P-450 Enzyme System/analysis , Drug Interactions , Hydroxycholecalciferols/antagonists & inhibitors , Hydroxycholecalciferols/pharmacology , Hydroxylation/drug effects , Streptomyces/enzymology , Streptomyces/isolation & purificationABSTRACT
In order to examine the role of the oxygen-derived free radicals in bone metabolism, the mouse calvaria organ culture system and the mouse bone marrow culture system were used. The activity of bone resorption stimulated by 1 alpha, 25(OH)2 vitamin D3 was inhibited by the addition of catalase, an eliminating agent of H2O2. Catalase also significantly suppressed the osteoclast-like cell formation induced by 1 alpha, 25(OH)2 vitamin D3 in a dose-dependent manner, whereas the agent had no effect on the mitogenic activity of the bone marrow-derived stromal cells. This suppression was reversed by adding H2O2 to the system. The effect of catalase was limited to the early stage precursor of the osteoclast-like cells. Moreover, when superoxide dismutase (SOD), which activates the production of H2O2, was added to the bone marrow culture, the number of osteoclast-like cells increased significantly in a dose-dependent manner. Finally, the source of the oxygen-derived free radicals was examined by measuring the fluorescence of 2.7-dichlorofluorescein (DCF), the obtained result indicated that the osteoclast-like cells could produce almost the equivalent amount of H2O2 as could the macrophages. These results strongly suggest that the oxygen-derived free radicals, particularly H2O2, may regulate bone resorption, promoting the differentiation of osteoclast precursor cells to osteoclasts.
Subject(s)
Bone Resorption , Hydrogen Peroxide , Osteoclasts/cytology , Animals , Bone Marrow Cells , Calcifediol/antagonists & inhibitors , Catalase/pharmacology , Cell Differentiation/drug effects , Cell Division/drug effects , Cells, Cultured , Free Radicals , Hydrogen Peroxide/metabolism , Male , Mice , Osteoclasts/metabolism , Superoxide Dismutase/pharmacologyABSTRACT
1. The elimination half-time of 25-hydroxyvitamin D in plasma was estimated after intravenous injection of the radioactively labelled metabolite in seven patients with primary hyperparathyroidism before and after excision of a parathyroid adenoma. 2. The elimination half-time of 25-hydroxyvitamin D was significantly shortened in primary hyperparathyroidism and reverted towards normal after parathyroidectomy. 3. The increased metabolic clearance of 25-hydroxyvitamin D in primary hyperparathyroidism was accounted for by an increased excretion of vitamin D-derived inactivation products in the faeces. 4. Enhanced hepatic inactivation of 25-hydroxyvitamin D may be important in the development of vitamin D deficiency in primary hyperparathyroidism.
