1,25-DIHYDROXYVITAMIN D-3 STIMULATES ADIPOCYTE DIFFERENTIATION IN CULTURES OF FETAL-RAT CALVARIA CELLS - COMPARISON WITH THE EFFECTS OF DEXAMETHASONE
1,25-DIHYDROXYVITAMIN D-3 STIMULATES ADIPOCYTE DIFFERENTIATION IN CULTURES OF FETAL-RAT CALVARIA CELLS - COMPARISON WITH THE EFFECTS OF DEXAMETHASONE
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DOI：
10.1210/en.134.5.2221
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发表时间：
1994-05-01
期刊：
影响因子：
4.8
通讯作者：
AUBIN, JE
中科院分区：
文献类型：
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作者：
BELLOWS, CG;WANG, YH;AUBIN, JE
几种间充质衍生细胞类型的祖细胞存在于新鲜分离的胎鼠颅盖 (RC) 细胞群中。我们表征了 1,25-二羟基维生素 D-3 [1,25-(OH)(2)D-3] 对原代 RC 细胞分化为脂肪细胞的影响，并将这些影响与地塞米松 (Dex) 的影响进行了比较。将 RC 细胞铺板于 3 x 10(4)/35-mm 培养皿中，并在含有 10% 胎牛血清、50 μg/ml 抗坏血酸、10 mM β-甘油磷酸钠和 0.1-100 nM 1,25-(OH)(2)D-3 或 1-1000 nM 的 α-最低必需培养基中维持培养物 14-19 天德克斯。使用形态学（苏丹 IV 染色培养物后脂肪细胞病灶数量和面积的定量）和生物化学（3-磷酸甘油脱氢酶活性）方法评估脂肪生成。在 1,25-(OH)(2)D-3 存在的情况下，汇合后约 3 天，脂肪细胞灶形成圆形或星状细胞簇。脂肪细胞灶发育的刺激在 0.1-100 nM 范围内具有剂量依赖性，并且在 10 nM 1,25-(OH)(2)D-3 时达到最大刺激；半最大刺激发生在约 1 nM 处。 1,25-(OH)(2)D-3 诱导的脂肪细胞刺激不需要抗坏血酸和 β-甘油磷酸的存在，但在 1,25-(OH)(2)D-3 存在下两者均显着增加脂肪细胞灶的数量。 1,25-(OH)(2)D-3启动脂肪细胞分化的关键期在1-9天之间，一旦沿着脂肪形成途径发生，脂肪细胞在没有1,25-(OH)(2)D-3的情况下保持其分化状态。 1,25-(OH)(2)D-3 的短期（48 小时）脉冲导致脂肪细胞形成轻微但显着的增加。其他维生素 D-3 代谢物在刺激脂肪细胞分化方面的效果不如 1,25-(OH)(2)D-3。 Dex (1-100 nM) 还引起 RC 细胞培养物中脂肪细胞灶分化的剂量依赖性增加。在存在 Dex 的情况下形成的脂肪细胞灶经常出现在培养物中较早的时间，即当细胞在第 6-7 天达到汇合时，并且比用 1,25-(OH)(2)D-3 形成的脂肪细胞灶更分散。然而，在混合 RC II-V 细胞中，1,25-(OH)(2)D-3 对脂肪细胞分化的刺激作用大于 Dex。 1,25-(OH)(2)D-3 和 Dex 的联合作用在低浓度下是相加的，在 1,25-(OH)(2)D-3 或 Dex 浓度较高时具有协同作用。数据显示，从胎儿 RC 中分离的骨细胞群含有脂肪细胞祖细胞，并且 1,25-(OH)(2)D-3 以及 Dex 是这些骨细胞群中脂肪细胞分化的有效调节剂。
Progenitor cells for several mesenchymally derived cell types exist within freshly isolated fetal rat calvaria (RC) cell populations. We have characterized the effects of 1,25-dihydroxyvitamin D-3 [1,25-(OH)(2)D-3] on the differentiation of adipocytes from primary RC cells and compared these effects with those of dexamethasone (Dex). RC cells were plated at 3 x 10(4)/35-mm dish, and cultures were maintained for 14-19 days in alpha-Minimum Essential Medium containing 10% fetal bovine serum, 50 mu g/ml ascorbic acid, 10 mM Na beta-glycerophosphate, and 0.1-100 nM 1,25-(OH)(2)D-3 or 1-1000 nM Dex. Morphological (quantitation of adipocyte foci number and area after staining cultures with Sudan IV) and biochemical (glycerol-3-phosphate dehydrogenase activity) methods of assessing adipogenesis were used. In the presence of 1,25-(OH)(2)D-3, adipocyte foci developed about 3 days after confluency as clusters of rounded or stellate cells. Stimulation of adipocyte foci development was dose dependent from 0.1-100 nM and was maximal with 10 nM 1,25-(OH)(2)D-3; half-maximal stimulation occurred at about 1 nM. The presence of ascorbic acid and beta-glycerophosphate was not required for 1,25-(OH)(2)D-3-induced stimulation of adipocytes, but both significantly increased the number of adipocyte foci in the presence of1,25-(OH)(2)D-3. The critical period for initiation of adipocyte differentiation with 1,25-(OH)(2)D-3 was between 1-9 days, and once committed along the adipogenic pathway, adipocytes maintained their differentiated state in the absence of 1,25-(OH)(2)D-3. Short term (48-h) pulses of 1,25-(OH)(2)D-3 resulted in slight, but significant, increases in adipocyte formation. Other vitamin D-3 metabolites were less effective than 1,25-(OH)(2)D-3 in stimulating adipocyte differentiation. Dex (1-100 nM) also caused a dose-dependent increase in the differentiation of adipocyte foci in RC cell cultures. The adipocyte foci that developed in the presence of Dex frequently appeared earlier in culture, i.e. when cells reached confluency on days 6-7, and were more diffuse than those forming with 1,25-(OH)(2)D-3. The stimulation of adipocyte differentiation by 1,25-(OH)(2)D-3, however, was greater than that by Dex in mixed RC II-V cells. The combined effects of 1,25-(OH)(2)D-3 and Dex were additive at low concentrations and synergistic at higher concentrations of either 1,25-(OH)(2)D-3 or Dex. The data show that bone cell populations isolated from fetal RC contain adipocyte progenitors and that 1,25-(OH)(2)D-3 as well as Dex are potent regulators of adipocyte differentiation within these bone cell populations.