24-25-dihydroxyvitamin-d-3 and Hypertrophy

Previously we showed that costochondral growth plate resting zone (RC) chondrocytes response primarily to 24R,25(OH)2D3 whereas prehypertrophic and hypertrophic (GC) cells respond to 1alpha,25(OH)2D3. 24R,25(OH)2D3 increases RC cell proliferation and inhibits activity of matrix processing enzymes, suggesting it stabilizes cells in the reserve zone, possibly by inhibiting the matrix degradation characteristic of apoptotic hypertrophic GC cells. To test this, apoptosis was induced in rat RC cells by treatment with exogenous inorganic phosphate (Pi). 24R,25(OH)2D3 blocked apoptotic effects in a dose-dependent manner. Similarly, apoptosis was induced in ATDC5 cell cultures and 24R,25(OH)2D3 blocked this effect. Further studies indicated that 24R,25(OH)2D3 acts via at least two independent pathways. 24R,25(OH)2D3 increases LPA receptor-1 (LPA R1) expression and production of lysophosphatidic acid (LPA), and subsequent LPA R1/3-dependent signaling, thereby decreasing p53 abundance. LPA also increases the Bcl-2/Bax ratio. In addition, 24R,25(OH)2D3 acts by increasing PKC activity. 24R,25(OH)2D3 stimulates 1-hydroxylase activity, resulting in increased levels of 1,25(OH)2D3, and it increases levels of phospholipase A2 activating protein, which is required for rapid 1alpha,25(OH)2D3-dependent activation of PKC in GC cells. These results suggest that 24R,25(OH)2D3 modulates growth plate development by controlling the rate and extent of RC chondrocyte transition to a GC chondrocyte phenotype.

Topics: 24,25-Dihydroxyvitamin D 3; Animals; Apoptosis; Calcitriol; Caspase 3; Cell Proliferation; Chondrocytes; Enzyme Activation; Growth Plate; Hypertrophy; Lysophospholipids; Male; Phospholipases A2; Rats; Rats, Sprague-Dawley; Tumor Suppressor Protein p53

In rabbit cartilage growth plates, the membrane potential, Vm, and potassium intracellular activities, alpha iK, were determined in order to study the effects of long-term (48 h) and short-term (1-2 min) exposures to vitamin D metabolites. Results are as follows: (i) in proliferative cells, Vm was -55.6 +/- 0.2 mV, n = 30, and alpha ik = 50.8 +/- 4.2 mM, n = 22; (ii) in hypertrophic cells, Vm was -35.5 +/- 0.8 mV, n = 88, and alpha iK = 85.1 +/- 5.4 mM, n = 20; (iii) Vm (-44.0 +/- 1.0 mV, n = 33) and alpha iK (114.7 +/- 8.7 mM, n = 14) were increased in metatarsal hypertrophic cells incubated with 10(-10)M of 1,25(OH)2D3 but were unaffected by the presence of 24,25(OH)2D3; (iv) an hyperpolarization of Vm was observed after short-term exposure of the hypertrophic cells to 10(-10)M of 1,25(OH)2D3 (-2.2 +/- 0.2 mV, n = 34) and 24,25(OH)2D3 (-2.2 +/- 0.7 mV, n = 17) but not to 25(OH)D3 (+0.3 +/- 0.8 mV, n = 10).

Topics: 24,25-Dihydroxyvitamin D 3; Animals; Calcitriol; Cartilage; Cell Division; Dihydroxycholecalciferols; Growth Plate; Hypertrophy; In Vitro Techniques; Male; Membrane Potentials; Microelectrodes; Ouabain; Potassium; Rabbits; Time Factors