Effects of vitamin D metabolites on healing of low phosphate, vitamin D-deficient induced rickets in rats

Atkin, I.; Pita, Julio C.; Ornoy, Asher; Agundez, Agueda; Castiglione, G.; Howell, David S.

doi:10.1016/8756-3282(85)90315-1

Cited by 53 publications

(16 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fracture healing in chickens has been recently shown to be enhanced by treatment with 24R,25-(OH) 2 D 3 (Kato et al, 1998). These observations support those of Atkin and colleagues (Atkin et al, 1985;Atkin et al, 1992) demonstrating an enhancement of repair of rickets after treatment with 24R,25-(OH) 2 D 3 . The importance of 24R,25-(OH) 2 D 3 to proper growth and development via endochondral ossification is also shown by the differential localization of [ 3 H]-24R,25-(OH) 2 D 3 in the growth plates of vitamin D-replete rats (Seo et al, 1996).…”

Section: Discussionsupporting

confidence: 58%

24R,25-(OH)2D3 mediates its membrane receptor-dependent effects on protein kinase C and alkaline phosphatase via phospholipase A2 and cyclooxygenase-1 but not cyclooxygenase-2 in growth plate chondrocytes

Schwartz¹,

Sylvia

Toro

et al. 2000

J. Cell. Phys.

View full text Add to dashboard Cite

Recent studies have shown that 24R,25-(OH)(2)D(3) mediates its effects on growth plate chondrocytes via membrane receptors. This study examined the roles of phospholipase A(2) (PLA(2)) and cyclooxygenase (Cox) in the mechanism of action of 24R, 25-(OH)(2)D(3) in resting zone chondrocytes in order to determine whether the activity of one or both enzymes provides a regulatory checkpoint in the signaling pathway resulting in increased protein kinase C (PKC) activity. We also determined whether constitutive or inducible Cox is involved. Cultures were incubated with 24R, 25-(OH)(2)D(3) for 90 min to measure PKC or for 24 h to measure physiological responses ([(3)H]-thymidine incorporation, alkaline phosphatase-specific activity, [(35)S]-sulfate incorporation). Based on RT-PCR and Northern blot analysis, resting zone chondrocytes express mRNAs for both Cox-1 and Cox-2. Levels of mRNA for both proteins were unchanged from control levels after a 24-h incubation with 24R,25-(OH)(2)D(3). To examine the role of Cox, the cultures were also treated with resveratrol (a specific inhibitor of Cox-1), NS-398 (a specific inhibitor of Cox-2), or indomethacin (a general Cox inhibitor). Cox-1 inhibition resulted in effects on proliferation, differentiation, and matrix production typical of 24R, 25-(OH)(2)D(3). In contrast, inhibition of Cox-2 had no effect, indicating that 24R,25-(OH)(2)D(3) exerts its effects via Cox-1. Inhibition of Cox-1 also blocked 24R,25-(OH)(2)D(3)-dependent increases in PKC. Activation of PLA(2) with melittin inhibited 24R, 25-(OH)(2)D(3)-dependent stimulation of PKC, and inhibition of PLA(2) with quinacrine stimulated PKC in response to 24R, 25-(OH)(2)D(3). Inclusion of resveratrol reduced the melittin-dependent inhibition of PLA(2) and caused an increase in quinacrine-stimulated PLA(2) activity. Metabolism of arachidonic acid to leukotrienes is not involved in the response to 24R, 25-(OH)(2)D(3) because inhibition of lipoxygenase had no effect. The effect of 24R,25-(OH)(2)D(3) was specific because 24S,25-(OH)(2)D(3), the biologically inactive stereoisomer, failed to elicit a response from the cells. These results support the hypothesis that 24R, 25-(OH)(2)D(3) exerts its effects via more than one signaling pathway and that these pathways are interrelated via the modulation of PLA(2). PKC regulation may occur at multiple stages in the signal transduction cascade.

show abstract

Section: Discussionsupporting

confidence: 58%

24R,25-(OH)2D3 mediates its membrane receptor-dependent effects on protein kinase C and alkaline phosphatase via phospholipase A2 and cyclooxygenase-1 but not cyclooxygenase-2 in growth plate chondrocytes

Schwartz¹,

Sylvia

Toro

et al. 2000

J. Cell. Phys.

View full text Add to dashboard Cite

show abstract

“…Whereas matrix vesicles produced by resting zone costochondral cartilage cell cultures are sensitive to 24,25-(OH),D3, those produced by growth zone cell cultures are sensitive to 1,25-(OH)zD3. Because maturation of the extracellular matrix by chondrocytes in the endochondral lineage is modulated'by these vitamin D metabolites in vivo (Atkin et al, 1985), we hypothesized, and eventually showed, that MMP activity in matrix vesicles is also regulated by 1,25-(OH)& and 24,25-(OH),D3 and that the effect of hormone is specific to the lineage stage of the chondrocyte, with resting zone cells preserving the proteoglycan content of the matrix and growth zone cells degrading this extracellular matrix component .…”

mentioning

confidence: 99%

Vitamin D3 regulation of stromelysin-1 (MMP-3) in chondrocyte cultures is mediated by protein kinase C

et al. 1996

View full text Add to dashboard Cite

Matrix metalloproteinases (MMPs) are a group of enzymes with the potential to degrade extracellular matrix proteins. One of the MMPs, stromelysin-1 (MMP-3) has been localized to extracellular matrix vesicles in growth plate chondrocyte cultures, suggesting involvement of this enzyme in remodeling of the extracellular matrix during endochondral development, a process which is regulated by the vitamin D metabolites, 1,25-(OH)2D3 and 24,25-(OH)2D3. To determine whether stromelysin-1 is regulated by vitamin D as well, confluent cultures of cells derived from growth zone (GC) and resting zone (RC) rat costochondral cartilage were treated with 1 alpha, 25-(OH)2D3 (1,25) and 24R,25-(OH)2D3 (24,25), respectively, and the effect on stromelysin-1 assessed by casein gel zymography and Western blots. Although stromelysin-1 activity was enriched in the matrix vesicle fraction, only the plasma membrane enzyme was affected by the treatment; 1, 25 and 24,25 caused a marked decrease in plasma membrane stromelysin-1 activity in their target cells. Since plasma membrane protein kinase C (PKC) activity is stimulated by 1,25 and 24,25, we hypothesized that stromelysin-1 activity was regulated by the vitamin D metabolites via PKC-dependent phosphorylation. To test this, membrane fractions (containing endogenous PKC alpha and zeta as well as stromelysin-1) were incubated in the presence of purified rat brain PKC and/or recombinant human (rh) stromelysin-1 and [gamma 32 P]-ATP and anti-stromelysin-1 immunoprecipitates were analyzed by autoradiography and Western blots. Immuno-phospho-stromelysin-1 was localized to a 52-kDa band in the plasma membrane fraction only; no phosphorylation was observed in the matrix vesicle fraction. Selective inhibitors of PKC activity demonstrated that phosphorylation was inhibited by H7 and low concentrations of H8, but not by HA1004, indicating that PKC, not PKA, was responsible. Protein phosphatase 2A1 (PP2A), a serine/threonine-specific phosphatase, selectively removed the radiolabel in a time-dependent manner, providing further support for a PKC-dependent phosphorylation mechanism. Incubation of resting zone cell plasma membranes with 24,25 but not 1, 25, resulted in phosphorylation of stromelysin-1, demonstrating that the nongenomic effect was metabolite-specific. This suggests that this may be one mechanism by which vitamin D metabolites regulate stromelysin-1 activity and that PKC-dependent phosphorylation inhibits the metalloproteinase.

show abstract

“…The present study with rachitic animals was designed to define how rat interstitial collagenase gene expression is regulated by the principal active metabolite of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25-(OH),D3), an important regulator of growth cartilage processes [Atkin et al, 1985;Kato et al, 1990;Schwartz and Boyan, 19881. Interleukin-lp (IL-lp) peptide is also hy-pothesized to mediate some actions of 1,25-(OH)2D3 in growth cartilage because IL-lp peptide increases fivefold in cartilage extracts after the addition of 1,25-(OH)2D3 [Howell et al, 19921.…”

mentioning

confidence: 99%

Regulation of rat interstitial collagenase gene expression in growth cartilage and chondrocytes by vitamin D3, interleukin‐1β, and okadaic acid

Grumbles¹,

Shao²,

Jeffrey³

et al. 1996

J. Cell. Biochem.

Self Cite

View full text Add to dashboard Cite

The interstitial collagenase produced by the rat growth plate chondrocytes is the homologue of the human collagenase-3, or matrix metalloproteinase-13. This enzyme is responsible for the loss of collagen during hypertrophy of chondrocytes and for the degradation of transverse septa in long bone growth. Rachitic rats (42 days, male Sprague-Dawley) had an 8-fold higher level of collagenase mRNA in the hypertrophic versus proliferative zone of growth plate cartilage. Intramuscular injection of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3; 1.0 micrograms/kg body weight) in rachitic rats increased collagenase mRNA another 1.5-fold in the hypertrophic zone. The regulation of collagenase gene by 1,25-(OH)2D3 and interleukin (IL)-1 beta in cultured proliferative chondrocytes was studied by means of steady-state mRNA and half-life determination of mRNA using the transcriptional inhibitor actinomycin D, and nuclear run-on transcription analyses. Treatment of cells with 1,25-(OH)2D3 (10(-6) M) and IL-1 beta (2 ng/ml) increased collagenase mRNA 8- and 13-fold, respectively. Additionally, the collagenase mRNA half-life was increased by 1,25-(OH)2D3 and IL-1 beta. In the presence of a protein kinase C inhibitor, staurosporine, 1,25-(OH)2 D3 induction of collagenase mRNA was blocked. Here the addition of phorbol 12-myrisate 13-acetate (PMA) to activate protein kinase C increased collagenase mRNA 10-fold. However, in the presence of staurosporine (50 nM), PMA induction was blocked, whereas IL-1 beta was not. IL-1 beta is known to activate several phosphorylation pathways. Okadaic acid (500 nM), a protein phosphatase inhibitor, increased the relative collagenase mRNA abundance 10-fold. The rate of the rat collagenase gene transcription in nuclei was increased with 1,25-(OH)2D3, IL-1 beta and okadaic acid. In separate experiments, the collagenase promoter was ligated to a reporter plasmid and the plasmid was transfected into chondrocytes. The results showed that 1,25-(OH)2D3, IL-1 beta, and PMA increased reporter activity 2.5-, 2.8-, and 3.27-fold, respectively. Thus, there are multiple nuclear and cytoplasmic mechanisms by which cartilage modulators regulate rat interstitial collagenase gene expression.

show abstract

Effects of vitamin D metabolites on healing of low phosphate, vitamin D-deficient induced rickets in rats

Cited by 53 publications

References 35 publications

24R,25-(OH)2D3 mediates its membrane receptor-dependent effects on protein kinase C and alkaline phosphatase via phospholipase A2 and cyclooxygenase-1 but not cyclooxygenase-2 in growth plate chondrocytes

24R,25-(OH)2D3 mediates its membrane receptor-dependent effects on protein kinase C and alkaline phosphatase via phospholipase A2 and cyclooxygenase-1 but not cyclooxygenase-2 in growth plate chondrocytes

Vitamin D3 regulation of stromelysin-1 (MMP-3) in chondrocyte cultures is mediated by protein kinase C

Regulation of rat interstitial collagenase gene expression in growth cartilage and chondrocytes by vitamin D3, interleukin‐1β, and okadaic acid

Contact Info

Product

Resources

About