Akt调节切应力诱导骨髓间充质干细胞Bmi-1基因的表达

发布时间：2018-01-21 00:42

本文关键词： 间充质干细胞基因表达流体切应力信号转导　出处：《医用生物力学》2017年01期 　论文类型：期刊论文

【摘要】：目的探讨流体切应力对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)中Bmi-1基因表达的影响及可能的信号机制。方法原代体外分离培养大鼠BMSCs,应用平行平板流动腔系统,给BMSCs施加不同强度(0.5、1.5、3.0 Pa)和不同加载时间(1、2、6、24 h)的层流切应力,用实时定量RT-PCR法检测Bmi-1基因的表达水平,用免疫印迹法检测磷酸化Akt和ERK1/2的表达水平,利用Wortmannin(PI3K特异性抑制剂)、PD98059(ERK1/2 MAPK特异性抑制剂)信号阻断剂探讨信号转导途径。结果 BMSCs在1.5 Pa切应力作用1 h后Bmi-1基因表达即明显增强,24 h达高峰。不同强度切应力都会刺激Bmi-1基因表达,其中3.0 Pa最强。切应力能显著激活磷酸化Akt和ERK1/2表达。Wortmannin而不是PD98059可以明显抑制Bmi-1基因的表达。结论切应力可诱导BMSCs中Bmi-1基因表达,其表达量与刺激时间和切应力的强度密切相关,这种作用可能通过Akt信号调节。
[Abstract]:Objective to investigate the effect of fluid shear stress on bone marrow marrow mesenchymal stem cells. Methods Rat BMSCs were isolated and cultured in vitro, and parallel plate flow chamber system was used. The laminar shear stress of BMSCs was applied to BMSCs with different strength of 0.5 ~ (1.5) Pa) and different loading time of 1 ~ 2 ~ 2 ~ 6 ~ 6 ~ (24 h). The expression levels of Bmi-1 gene and phosphorylated Akt and ERK1/2 were detected by real-time quantitative RT-PCR and Western blot. Wortmannin(PI3K specific inhibitors were used. PD98059(ERK1/2 MAPK specific inhibitor. Results the expression of Bmi-1 gene increased significantly after 1 h of BMSCs was exposed to 1.5 Pa shear stress. The peak was reached at 24 h. The Bmi-1 gene expression was stimulated by different shear stress. Of which 3.0. Pa is the strongest. Shear stress can significantly activate the expression of phosphorylated Akt and ERK1/2. Wortmannin, instead of PD98059, can significantly inhibit the expression of Bmi-1 gene. Conclusion Shear stress can significantly inhibit the expression of Bmi-1 gene. It can induce the expression of Bmi-1 gene in BMSCs. The expression level is closely related to the stimulation time and shear stress, which may be regulated by Akt signal.
【作者单位】：湖北医药学院附属随州医院转化医学研究中心;湖北医药学院基础医学院;
【基金】：国家自然科学基金项目(31670961) 湖北省科技厅自然基金项目(2015CFB186) 湖北医药学院大学生创新创业训练计划项目(2015XS10)
【分类号】：R329.2
【正文快照】： 骨髓间充质干细胞(bone mesenchymal stemcells,BMSCs)是一种来源于中胚层的多能干细胞,具有多向分化潜能。干细胞在体内生长的局部微环境(包括细胞外基质、细胞因子在内的基质微环境和复杂的力学微环境共同组成)对其分化走向起重要的调控作用[1-2]。流体切应力(fluid shear s

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