Roles of brain and muscle ARNT-like 1 and Wnt antagonist Dkk1 during osteogenesis of bone marrow stromal cells
Roles of brain and muscle ARNT-like 1 and Wnt antagonist Dkk1 during osteogenesis of bone marrow stromal cells
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DOI：
10.1111/cpr.12075
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发表时间：
2013-12-01
影响因子：
8.5
通讯作者：
Tan, Z.
中科院分区：
文献类型：
--
作者：
He, Y.;Chen, Y.;Tan, Z.
目的许多研究表明，生物钟基因--脑和肌肉ARNT样1（Bmal 1）通过影响间充质干细胞（MSCs）的年龄相关变化与骨衰老直接相关。Wnt作为一种主要的发育信号，可能在这一过程中发挥重要作用。在这里，我们的目的是阐明是否Bmal 1积极调节osteogenesi通过Wnt pathways.Materials and methodsBone marrow stromal cells cultured in basic and in osteo-induction medium with Wnt signaling inhibitor Dkk 1 and Bmal 1 transfection.通过流式细胞术、衰老相关半乳糖苷酶（SA-gal）染色、实时定量PCR和westernblot分析，观察骨髓间充质干细胞的增殖和成骨能力、Bmal 1的表达和Wnt信号通路的激活情况。当通过转染增加细胞Bmal 1时，通过激活Wnt信号在一定程度上挽救了Dkk 1对成骨的抑制。然而，Dkk 1并没有显着影响增殖或衰老的MSC在早期culture.ConclusionThese研究结果表明，Bmal 1和Wnt信号可能有协同作用，在特定阶段的成骨。抑制Wnt信号传导并没有通过早期传代极大地影响MSC的老化。
ObjectivesMany studies have demonstrated that the clock gene, brain and muscle ARNT-like 1 (Bmal1), is directly related to bone ageing by affecting age-related changes to mesenchymal stem cells (MSCs). As a main developmental signal, Wnt may play an important role in this process. Here, we have aimed to elucidate whether Bmal1 positively regulates osteogenesi via Wnt pathways.Materials and methodsBone marrow stromal cells were cultured in basic and in osteo-induction medium with Wnt signalling inhibitor Dkk1 and Bmal1 transfection. Proliferation and osteogenesis of MSCs, expression of Bmal1 and activation of Wnt signalling were investigated by flow cytometry, senescence-associated -galactosidase (SA--gal) staining, real-time quantitative PCR and western blot analysis.ResultsExpression of Bmal1 (specially after 7days osteo-induction), activation of Wnt signalling and osteo-related factors fell significantly during osteo-induction after Dkk1 addition. When cellular Bmal1 was increased by transfection, osteogenesis inhibition by Dkk1 was rescued to a certain extent with activation of Wnt signalling. However, Dkk1 did not significantly affect proliferation or senescence of MSCs during early periods of culture.ConclusionThese findings demonstrated that Bmal1 and Wnt signalling may have a synergistic effect at a particular stage of osteogenesis. Inhibition of Wnt signalling did not greatly affect ageing of MSCs through early passages.