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| orexin-1受体抑制剂通过Wnt通路促进骨髓间充质干 细胞的成骨分化 |
| In vitro study on the effect of Wnt signaling pathway on orexin-1 receptor inhibitor induced osteogenic differentiation of rat bone marrow mesenchymal stem cells |
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| DOI：10.3969/j.issn.1006-7108.2017.05.010 |
| 中文关键词: 骨髓间充质干细胞；Writ/β-catenin信号通路 成骨分化 |
| 英文关键词:Bone marrow mesenchymal stem cells Wnt/beta-catenin signaling pathways Osteogenic differentiation |
| 基金项目:2014年广东医学院科研基金（2XJ141P)；2015年湛江市非资助科技攻关计划项目（2015B01083)；2015年湛江市资助科技攻关计划（2015A01030） |
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| 中文摘要: |
| 目的 研究Wnt/β-catenin信号通路对orexin-1受体抑制剂介导大鼠骨髓间充质干细胞（BMSCs)成骨分化的影响。方 法（1)分离提取SD大鼠股骨、胫骨的BMSCs,用流式细胞仪进行细胞表型鉴定；（2)实验分组：对照组、10–5 mol/L、10-6 mol/L质量浓度的orexin-1受体抑制剂溶液，于成骨诱导第5天免疫印迹法和实时荧光定量PCR法观察Wnt/β-catenin信号通 路关键靶点蛋白Dickkopf-1、Gsk3β、β-catenin以及基因Gsk3β、β-catenin mRNA表达，以观察Wnt信号通路对orexin-1受体抑 制剂诱导大鼠骨髓间充质干细胞成骨分化的影响。结果10–5 mol/L、10-6 mol/L的orexin-1受体抑制剂处理BMSCs5天后， Dickkopf-1、β-catenin和GSK3β蛋白表达升高，β-catenin和GSK3β mRNA表达升高。结论orexin-1受体抑制剂促进大鼠骨髓 基质干细胞向成骨细胞方向分化的作用可能与调控Wnt/β-catenin信号通路有关。 |
| 英文摘要: |
| Objective To observe the effect of Wnt/intracellular beta chain protein (β-catenin) signaling pathway on orexin-1 receptor inhibitor induced osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in rats. Methods (1) Isolated the bone marrow mesenchymal stem cells from one-month SD rats by whole bone marrow culture method and identified the cell surface antigen by flow cytometry. (2) Experimental groups： control group, and 10–5 mol/L or 10–6 mol/L orexin-1 receptor inhibitor solution groups. At day 5 the mRNA expression of Gsk3beta and beta-catenin were detected by real-time quantitative PCR. The protein expression of Dickkopfl, Gsk3beta and beta-catenin were detected by Western blot. Results Orexin-1 receptor inhibitor up-regulated Gsk3beta and beta-catenin mRNA expression at concentrations of 10– 6 mol/L and 10 –5 mol/L for 5 days. Orexin-1 receptor inhibitor up-regulated Dickkopfl，Gsk3beta and beta-catenin protein expression at concentrations of 10–6 mol/L and 10 –5 mol/L for 5 days (P <
0.05 ). Conclusion The effect of orexin-1 receptor inhibitor on osteogenic differentiation of MSCs maybe closely related to the regulation of Wnt/catenin signaling pathway. |
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