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原代人成骨细胞对25-羟基维生素D3、1,25-二羟基维生素D3和24R,25-二羟基维生素D3的反应。

Primary human osteoblasts in response to 25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3 and 24R,25-dihydroxyvitamin D3.

作者信息

van der Meijden Karen, Lips Paul, van Driel Marjolein, Heijboer Annemieke C, Schulten Engelbert A J M, den Heijer Martin, Bravenboer Nathalie

机构信息

Department of Internal Medicine/Endocrinology, VU University Medical Center, Research Institute MOVE, Amsterdam, The Netherlands.

Department of Internal Medicine/Endocrinology, Erasmus Medical Center, Rotterdam, The Netherlands.

出版信息

PLoS One. 2014 Oct 17;9(10):e110283. doi: 10.1371/journal.pone.0110283. eCollection 2014.

DOI:10.1371/journal.pone.0110283
PMID:25329305
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4201491/
Abstract

The most biologically active metabolite 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has well known direct effects on osteoblast growth and differentiation in vitro. The precursor 25-hydroxyvitamin D3 (25(OH)D3) can affect osteoblast function via conversion to 1,25(OH)2D3, however, it is largely unknown whether 25(OH)D3 can affect primary osteoblast function on its own. Furthermore, 25(OH)D3 is not only converted to 1,25(OH)2D3, but also to 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) which may have bioactivity as well. Therefore we used a primary human osteoblast model to examine whether 25(OH)D3 itself can affect osteoblast function using CYP27B1 silencing and to investigate whether 24R,25(OH)2D3 can affect osteoblast function. We showed that primary human osteoblasts responded to both 25(OH)D3 and 1,25(OH)2D3 by reducing their proliferation and enhancing their differentiation by the increase of alkaline phosphatase, osteocalcin and osteopontin expression. Osteoblasts expressed CYP27B1 and CYP24 and synthesized 1,25(OH)2D3 and 24R,25(OH)2D3 dose-dependently. Silencing of CYP27B1 resulted in a decline of 1,25(OH)2D3 synthesis, but we observed no significant differences in mRNA levels of differentiation markers in CYP27B1-silenced cells compared to control cells after treatment with 25(OH)D3. We demonstrated that 24R,25(OH)2D3 increased mRNA levels of alkaline phosphatase, osteocalcin and osteopontin. In addition, 24R,25(OH)2D3 strongly increased CYP24 mRNA. In conclusion, the vitamin D metabolites 25(OH)D3, 1,25(OH)2D3 and 24R,25(OH)2D3 can affect osteoblast differentiation directly or indirectly. We showed that primary human osteoblasts not only respond to 1,25(OH)2D3, but also to 24R,25(OH)2D3 by enhancing osteoblast differentiation. This suggests that 25(OH)D3 can affect osteoblast differentiation via conversion to the active metabolite 1,25(OH)2D3, but also via conversion to 24R,25(OH)2D3. Whether 25(OH)D3 has direct actions on osteoblast function needs further investigation.

摘要

生物活性最强的代谢产物1,25 - 二羟基维生素D3（1,25(OH)2D3）在体外对成骨细胞的生长和分化具有众所周知的直接作用。前体25 - 羟基维生素D3（25(OH)D3）可通过转化为1,25(OH)2D3来影响成骨细胞功能，然而，2,5(OH)D3自身是否能影响原代成骨细胞功能在很大程度上尚不清楚。此外，25(OH)D3不仅会转化为1,25(OH)2D3，还会转化为可能同样具有生物活性的24R,25 - 二羟基维生素D3（24R,25(OH)2D3）。因此，我们使用原代人成骨细胞模型，通过沉默CYP27B1来研究25(OH)D3自身是否能影响成骨细胞功能，并探究24R,25(OH)2D3是否能影响成骨细胞功能。我们发现，原代人成骨细胞对25(OH)D3和1,25(OH)2D3均有反应，表现为增殖减少，且通过碱性磷酸酶、骨钙素和骨桥蛋白表达增加来增强分化。成骨细胞表达CYP27B1和CYP24，并呈剂量依赖性地合成1,25(OH)2D3和24R,25(OH)2D3。沉默CYP27B1导致1,25(OH)2D3合成减少，但在用25(OH)D3处理后，与对照细胞相比，我们在CYP27B1沉默细胞中未观察到分化标志物mRNA水平的显著差异。我们证明，24R,25(OH)2D3可增加碱性磷酸酶、骨钙素和骨桥蛋白的mRNA水平。此外，24R,25(OH)2D3能显著增加CYP24 mRNA。总之，维生素D代谢产物25(OH)D3、1,25(OH)2D3和24R,25(OH)2D3可直接或间接影响成骨细胞分化。我们发现，原代人成骨细胞不仅对1,25(OH)2D3有反应，对24R,25(OH)2D3也有反应，可通过增强成骨细胞分化来响应。这表明，25(OH)D3可通过转化为活性代谢产物1,25(OH)2D3来影响成骨细胞分化，也可通过转化为24R,25(OH)2D3来实现。25(OH)D3对成骨细胞功能是否具有直接作用有待进一步研究。

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