CREB/Wnt10b mediates the effect of COX-2 on promoting BMP9-induced osteogenic differentiation via reducing adipogenic differentiation in mesenchymal stem cells
CREB/Wnt10b mediates the effect of COX-2 on promoting BMP9-induced osteogenic differentiation via reducing adipogenic differentiation in mesenchymal stem cells
复制标题
CREB/Wnt10b通过减少间充质干细胞的成脂分化介导COX-2促进BMP9诱导的成骨分化的作用
DOI：
10.1002/jcb.28234
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发表时间：
2019-06-01
影响因子：
4
通讯作者：
He, Bai-Cheng
中科院分区：
文献类型：
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作者：
Liao, Yun-Peng;Du, Wei-Min;He, Bai-Cheng
骨形态发生蛋白9 （Bone morphogenetic protein 9, BMP9）是最有效的成骨因子之一，可能是骨组织工程的潜在候选因子。然而，BMP9的成骨能力仍需进一步增强。在这项研究中，我们确定了Wnt10b对bmp9诱导的间充质干细胞（MSCs）成骨分化的影响以及这一过程的可能机制。我们采用聚合酶链反应（PCR）、Western blot分析、组织化学染色、异位骨形成和显微计算机断层扫描分析来评估Wnt10b对bmp9诱导的成骨分化的影响。同时，采用PCR、Western blot、染色质免疫沉淀、免疫沉淀等方法分析BMP9与Wnt10b可能存在的关系。我们发现BMP9上调C3H10T1/2细胞中的Wnt10b。Wnt10b增加了C3H10T1/2细胞中BMP9诱导的成骨标志物和骨形成，沉默Wnt10b可降低BMP9的这些作用。同时，Wnt10b可提高BMP9诱导的磷酸化Smad1/5/8 （p-Smad1/5/8）水平，而沉默Wnt10b可降低这一水平。相反，Wnt10b抑制BMP9诱导的脂肪生成标记物，沉默Wnt10b可以降低脂肪生成标记物。进一步分析表明，BMP9同时上调环氧化酶-2 （COX-2）和camp响应元件结合（p-CREB）的磷酸化。COX-2增强了BMP9增加p-CREB和Wnt10b的作用，而沉默COX-2则降低了这些作用。p-CREB与p-Smad1/5/8相互作用，结合C3H10T1/2细胞中Wnt10b的启动子。我们的研究结果表明，Wnt10b可以促进BMP - 9诱导的MSCs成骨分化，这可能是通过增强BMP/Smad信号，减少成脂分化介导的；BMP9可能通过COX-2/p- creb依赖的方式上调Wnt10b。
Bone morphogenetic protein 9 (BMP9) is one of the most potent osteogenic factors, which may be a potential candidate for bone tissue engineering. However, the osteogenic capacity of BMP9 still need to be further enhanced. In this study, we determined the effect of Wnt10b on BMP9-induced osteogenic differentiation in mesenchymal stem cell (MSCs) and the possible mechanism underlying this process. We introduced the polymerase chain reaction (PCR), Western blot analysis, histochemical stain, ectopic bone formation, and microcomputed tomography analysis to evaluate the effect of Wnt10b on BMP9-induced osteogenic differentiation. Meanwhile, PCR, Western blot analysis, chromatin immunoprecipitation, and immunoprecipitation were used to analyze the possible relationship between BMP9 and Wnt10b. We found that BMP9 upregulates Wnt10b in C3H10T1/2 cells. Wnt10b increases the osteogenic markers and bone formation induced by BMP9 in C3H10T1/2 cells, and silencing Wnt10b decreases these effects of BMP9. Meanwhile, Wnt10b enhances the level of phosphorylated Smad1/5/8 (p-Smad1/5/8) induced by BMP9, which can be reduced by silencing Wnt10b. On the contrary, Wnt10b inhibits adipogenic markers induced by BMP9, which can be decreased by silencing Wnt10b. Further analysis indicated that BMP9 upregulates cyclooxygenase-2 (COX-2) and phosphorylation of cAMP-responsive element binding (p-CREB) simultaneously. COX-2 potentiates the effect of BMP9 on increasing p-CREB and Wnt10b, while silencing COX-2 decreases these effects. p-CREB interacts with p-Smad1/5/8 to bind the promoter of Wnt10b in C3H10T1/2 cells. Our findings suggested that Wnt10b can promote BMP9-induced osteogenic differentiation in MSCs, which may be mediated through enhancing BMP/Smad signal and reducing adipogenic differentiation; BMP9 may upregulate Wnt10b via the COX-2/p-CREB-dependent manner.