THE EFFECTS OF VITAMIN-D METABOLITES ON THE PLASMA AND MATRIX VESICLE MEMBRANES OF GROWTH AND RESTING CARTILAGE CELLS-INVITRO
THE EFFECTS OF VITAMIN-D METABOLITES ON THE PLASMA AND MATRIX VESICLE MEMBRANES OF GROWTH AND RESTING CARTILAGE CELLS-INVITRO
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DOI：
10.1210/endo-122-6-2851
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发表时间：
1988-06-01
期刊：
影响因子：
4.8
通讯作者：
RAMIREZ, V
中科院分区：
文献类型：
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作者：
BOYAN, BD;SCHWARTZ, Z;RAMIREZ, V
本研究比较了维生素1,25-二羟基维生素D3 [1,25-(OH)2D3]和24,25-(OH)2D3对不同发育阶段软骨细胞群体的影响。取大鼠肋软骨静息区和邻近生长期的三代软骨细胞，在含有10%胎牛血清和增加激素浓度的Dulbecco' s Modified Eagle培养基中培养。测定细胞数量后，差速离心分离基质囊泡和质膜。激素对碱性磷酸酶、5′-核苷酸酶、瓦阿巴因敏感的Na+/K+- atp酶和磷脂组成的影响依赖于维生素D代谢物，并且具有细胞特异性。生长软骨细胞主要对1,25-(OH)2D3起反应，而静息区细胞主要对24,25-(OH)2D3起反应。1,25-(OH)2D3在药理学浓度下抑制生长软骨细胞数量，对静息软骨细胞数量无影响。相比之下，24,25-(OH)2D3在生理浓度下似乎可以刺激静息软骨细胞数量，在药理学剂量下可以抑制这些细胞，但对软骨细胞的生长没有影响。这些数据得到了[3H]胸苷结合研究的支持。1,25-(OH)2D3刺激生长软骨细胞基质囊泡中的碱性磷酸酶、5′-核苷酸酶活性和Na+/K+- atp酶活性。1,25-(OH)2D3还能刺激静息区细胞基质囊泡和质膜中的Na+/K+- atp酶活性。24,25-(OH)2D3刺激静息区细胞产生的基质囊泡中的碱性磷酸酶、5′-核苷酸酶和Na+/K+ atp酶。此外，24,25-(OH)2D3刺激了静息区细胞质膜以及生长软骨细胞基质囊泡和质膜中的Na+/K+- atp酶活性。
This study compares the effects of vitamins 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] and 24,25-(OH)2D3 on populations of chondrocytes at different developmental stages. Confluent third passage chondrocytes derived from the resting zone and adjacent growth region of rat costochondral cartilage were cultured in Dulbecco''s Modified Eagle''s Medium containing 10% fetal bovine serum and increasing concentrations of hormone. After determination of cell number, matrix vesicles and plasma membranes were isolated by differential centrifugation. The effects of hormone on alkaline phosphatase, 5''-nucleotidase, ouabain-sensitive Na+/K+-ATPase, and phospholipid composition were dependent on vitamin D metabolite and were cell specific. Growth cartilage chondrocytes responded primarily to 1,25-(OH)2D3, whereas resting zone cells responded primarily to 24,25-(OH)2D3. 1,25-(OH)2D3 inhibited growth cartilage cell number at pharmacological concentrations and had no effect on resting cartilage cell number. In contrast, 24,25-(OH)2D3 appeared to stimulate resting cartilage cell number at physiological concentrations and inhibit these cells at pharmacological doses, but had no effect on growth cartilage chondrocytes. These data were supported by [3H]thymidine incorporation studies. 1,25-(OH)2D3 stimulated alkaline phosphatase, 5''-nucleotidase activity, and Na+/K+-ATPase activity in the matrix vesicles of growth cartilage cells. 1,25-(OH)2D3 also stimulated Na+/K+-ATPase activity in the matrix vesicles and plasma membranes of resting zone cells. Incubation with 24,25-(OH)2D3 stimulated alkaline phosphatase, 5''-nucleotidase, and Na+/K+ ATPase in the matrix vesicles produced by resting zone cells. In addition, 24,25-(OH)2D3 stimulated Na+/K+-ATPase activity in the plasma membranes of resting zones cells as well as in both matrix vesicles and plasma membranes of growth cartilage cells.