| 喻伟,程鹏,黄晖,等.感觉神经肽P物质激活Wnt/β-catenin信号通路促进间充质干细胞增殖的实验研究.骨科,2015,6(2): 62-65. |
| 感觉神经肽P物质激活Wnt/β-catenin信号通路促进间充质干细胞增殖的实验研究 |
| Neuropeptide SP activates the Wnt/β-catenin signaling pathway and enhances the proliferation of bone marrow mesenchymal stem cells |
| 投稿时间：2014-08-14 |
| DOI：10.3969/j.issn.1674-8573.2015.02.002 |
| 中文关键词: 感觉神经肽P物质 骨髓间充质干细胞 增殖 Wnt/β-catenin信号通路 |
| 英文关键词: Substance P Bone marrow mesenchymal stem cells Proliferation Wnt/β-catenin signaling pathway |
| 基金项目:国家自然科学基金资助项目（No.81171696）；湖北省医学领军人才培养工程专项经费资助项目 |
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| 中文摘要: |
| 目的 探讨感觉神经肽P物质（substance P, SP）对骨髓间充质干细胞（bone marrow mesenchymal stem cells, BMSCs）增殖的影响并初步揭示其中可能的机制。方法 体外培养SD（Sprague-Dawley）大鼠BMSCs。细胞增殖-毒性检测试剂盒(Cell Counting Kit-8，CCK-8)法检测不同浓度SP刺激下的BMSCs增殖活性；在10-8 mol/L的SP刺激下，采用实时荧光定量PCR（real-time qPCR，RT-qPCR）检测第1、3、5和7 天时Wnt/β-catenin信号通路相关基因（β-catenin、c-myc、cyclin D）的表达情况。SP组单纯添加10-8 mol/L的SP，SP+LiCl（糖原合成激酶-3的抑制剂LiCl）组在SP组基础上添加6×10-3 mol/L的LiCl，SP+Dkk-1 (Dickkopf-1)组在SP组基础上添加10-9 mol/L的Dkk-1，以添加等量聚丁烯琥珀酸酯(poly butylenes succinate，PBS)为空白对照组。结果 CCK-8法显示SP明显促进BMSCs增殖，以10-8 mol/L的SP作用最为显著。RT-qPCR结果显示SP刺激下，β-catenin、c-myc、cyclin D的mRNA表达有显著差异，10-8 mol/L的浓度差异最为显著，并且呈明显的时间依赖，第5天mRNA表达出现顶峰。并且SP+LiCl组β-catenin、c-myc、cyclin D的mRNA和蛋白水平显著增强，而SP+Dkk-1组明显抑制。结论 SP通过激活Wnt/β-catenin通路而显著促进BMSCs增殖。 |
| 英文摘要: |
| Objective To explore the effects of neuropeptide substance P (SP) on proliferation of bone marrow mesenchymal stem cells (BMSCs) and reveal the potential mechanism. Methods BMSCs of SD rats were cultured at 37 ℃ and 5% CO2. The proliferative ability of BMSCs with different doses of SP was measured by CCK-8 kit. The expression of the related genes of the Wnt/β-catenin signaling pathway, such as β-catenin, c-myc, cyclin D, was detected by RT-qPCR in the dose-course and time-course. In addition, the agonist LiCl and the inhibitor Dkk-1 of the Wnt/β-catenin signaling pathway were separately used and the expression of β-catenin, c-myc, and cyclin D mRNA and proteins was detected as well. Results The BMSCs staining was typical with alizarin red staining after inducion of osteogenesis, oil red O staining after induction of adipogenesis, and alcian blue staining after induction of chondrogenesis. The BMSCs treated different doses of SP, especially 10-8 mol/L, had a greater proliferative ability than the controls. The mRNA expression of β-catenin, c-myc, and cyclin D was significantly different in SP dose-course and time-course, especially in 10-8 mol/L after 5 days. The expression levels of related genes to the Wnt/β-catenin signaling pathway were up-regulated by LiCl, and down-regulated by Dkk-1. Conclusion Neuropeptide SP can enhance the proliferation of BMSCs probably by regulating the Wnt/β-catenin signaling pathway. |
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