The vitamin D receptor regulates miR-140-5p and targets the MAPK pathway in bone development
The vitamin D receptor regulates miR-140-5p and targets the MAPK pathway in bone development
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维生素 D 受体调节 miR-140-5p 并靶向骨发育中的 MAPK 通路。
DOI：
10.1016/j.metabol.2018.03.018
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发表时间：
2018-08-01
影响因子：
9.8
通讯作者：
Zhao, Qun
中科院分区：
文献类型：
--
作者：
Luo, Wenting;Liu, Lingli;Zhao, Qun
背景：骨骼发育是一个复杂的过程。在胚胎期，维生素D （VD）的状态与胎儿骨骼发育密切相关。最近，在一些物种中发现mirna参与骨骼生长发育的调控。然而，在骨骼发育过程中，维生素D及其受体（VDR）和mirna之间相互作用的机制尚不清楚。本研究的目的是鉴定小鼠成骨细胞中受1,25(OH)(2)D-3调控的miRNAs，并分析VD/VDR与miRNAs在体外和体内的关系和作用。方法：我们对小鼠原代成骨细胞和经1,25(OH)(2)D-3处理的成骨细胞系进行了miRNA测序，以鉴定这些细胞中的miRNA。qRT-PCR验证后，选择miR-140-5p进行进一步分析。我们评估了包括miR-140-5p在内的几种表达mirna的预测靶基因的通路，通过qRT-PCR验证了MAPK通路中预测的靶基因，并在体外和体内探索了VD/VDR与miR-140-5p之间的相关性。结果：小鼠原代成骨细胞中有88个mirna表达差异，成骨细胞系中有49个mirna表达差异。MiR-140-5p在这两种小鼠成骨细胞中表达上调。1,25(OH)(2)D-3通过VDR转录激活促进miR-140-5p的表达，靶向抑制成骨细胞中的MAPK信号。在VDR敲除小鼠和165个人血清标本中观察到维生素D/VDR和miR-140-5p之间的正相关。这些数据首次表明VDR转录激活miR-140-5p。因此，VD/VDR/miR-140-5p/MAPK信号轴在传递1,25(OH)(2)D-3的作用中起重要作用。结论：我们的研究结果表明，miR-140-5p在体外和体内通过VD/VDR靶向MAPK通路是一种新的调控机制。这些发现为维生素d相关骨病的机制研究和治疗途径提供了新的参考。(C) 2018爱思唯尔公司版权所有。
Background: Skeletal development is a complicated process. The status of vitamin D (VD) is closely related to fetal bone development in the embryonic period. Recently, miRNAs have been found to participate in the regulation of skeletal growth and development in several species. However, the mechanisms underlying the interactions among vitamin D, its receptor (VDR), and miRNAs during the process of bone development remain unclear. The aim of this study was to identify miRNAs that are regulated by 1,25(OH)(2)D-3 in murine osteoblasts and to analyze the relationship and the effects of VD/VDR and miRNAs in vitro and in vivo.Methods: We performed miRNA sequencing in murine primary osteoblasts and in an osteoblast cell line treated with 1,25(OH)(2)D-3 to identify miRNAs in these cells. After qRT-PCR validation, miR-140-5p was selected for further analysis. We assessed the pathways comprising predicted target genes for several expressed miRNAs, including miR-140-5p, validated predicted target genes in the MAPK pathway by qRT-PCR, and explored the correlation between VD/VDR and miR-140-5p in vitro and in vivo.Results: 88 miRNAs in murine primary osteoblasts and 49 miRNAs in osteoblast cell line were found to be differentially expressed. MiR-140-5p was upregulated in these 2 types of murine osteoblasts. The expression of miR-140-5p was promoted by 1,25(OH)(2)D-3 through transcriptional activation by VDR, with targeted inhibition of MAPK signaling in osteoblasts. A positive correlation between vitamin D/VDR and miR-140-5p was observed in VDR-knockout mice and in 165 human serum specimens. These data show for the first time that VDR transcriptionally activates miR-140-5p. Therefore, the VD/VDR/miR-140-5p/MAPK signaling axis plays an important role in transmitting the effects of 1,25(OH)(2)D-3.Conclusion: Our results demonstrate a novel regulatory mechanism by which miR-140-5p targets the MAPK pathway by means of VD/VDR in vitro and in vivo. These findings provide a new reference for mechanistic research and therapeutic approaches for vitamin D-related bone diseases. (C) 2018 Elsevier Inc. All rights reserved.