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| 骨细胞Wnt/β-Catenin通过Notch信号促进BMSCs成骨分化 |
| Wnt/β-Catenin induces osteoblastic differentiation of BMCS via Notch signaling pathway by osteocytes |
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| DOI：10.3969/j.issn.1006.7108.2018.05.008 |
| 中文关键词: 骨细胞 Wnt/β-Catenin 骨髓间充质干细胞 Notch 成骨分化 |
| 英文关键词:Osteocytes Wnt/β-Catenin BMSCs Notch Osteogenic differentiation |
| 基金项目:国家自然科学基金( 81371972, 81572142) |
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| 中文摘要: |
| 目的 探讨骨细胞Wnt信号通路对骨髓间充质干细胞（BMSCs）成骨分化的作用及其分子机制。 方法 采用条件性基因敲出Cre/loxp技术制备特异性激活骨细胞Wnt/β-Catenin信号通路的小鼠；体外分离其和野生对照组小鼠的骨细胞，并分别与野生型小鼠骨髓间充质干细胞共培养；碱性磷酸酶（ALP）染色及活性定量、茜素红（Alizarin red）染色检测共培养早期成骨分化和晚期钙盐沉积；Real-Time PCR检测骨细胞Notch信号配体Jag1、Jag2、Dll1、Dll4及共培养后成骨分化特异标志物Runx2、ALP、Osteocalcin的mRNA表达水平。随后于共培养中加入Notch信号通路化学抑制剂DAPT（对照组加DMSO），再次行碱性磷酸酶（ALP）染色及活性定量和Real-Time PCR检测成骨分化特异标志物表达水平。结果 Wnt/β-Catenin激活的骨细胞其自身Jag1、Jag2、Dll4 mRNA表达较对照组骨细胞明显升高（P<0.05），与BMSCs共培养后成骨转录因子Runx2，成骨分化特异标志物ALP、Osteocalcin mRNA的表达较野生型明显升高（P<0.05），钙盐沉积增加。加入DAPT后，骨细胞Wnt激活组成骨转录因子Runx2、ALP、Osteocalcin mRNA的表达明显下降（P<0.05）。结论 骨细胞调控骨的代谢，激活其Wnt/β-Catenin信号通路将通过上调Notch信号而促进BMSCs的成骨分化。 |
| 英文摘要: |
| Objective To determine the effect of osteocytic Wnt/β-Catenin signaling on osteoblastic differentiation in bone marrow stromal cells (BMSCs) and its molecular mechanism. Methods Mice with osteocytic Wnt activation (daβcatOt) were generated previously by crossing DMP1-8kb-Cre mice with catnblox(ex3) mice. The osteocytes were isolated from Wnt activation mice and wild type mice. Then the two types osteocytes were separately co-cultured with wild-type BMSCs. ALP staining and Alizarin red staining were performed to measure osteoblastic differentiation in co-culture. The expressions of osteocytic Notch ligands Jag1, Jag2, Dll1, and Dll4 and osteoblastic marker genes Runx2, ALP, and osteocalcin were detected by qPCR. Furthermore, Notch signaling inhibitor DAPT was applied to the co-culture and osteoblastic marker genes were detected by qPCR again. Results The expression of Jag1, Jag2, and D114 mRNA in daβcatOt osteocytes was significantly higher than that in the control (P<0.05). In the co-cultures, the expression of osteoblastic makers Runx2, ALP, and osteocalcin mRNA was significantly higher than that in the control (P<0.05), and the deposition of calcium also increased compared to the control. The expression of osteoblastic transcription factors Runx2, ALP and Osteocalcin mRNA decreased significantly after addition of Notch inhibitor DAPT (P<0.05). Conclusion Osteocyte regulates bone metabolism. The activation of Wnt/β-Catenin signaling pathway in osteocytes promotes osteogenic differentiation of BMSCs via upregulation of Notch signaling. |
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