机械牵张力促进小鼠骨髓间充质干细胞的成骨向分化

发布时间：2018-09-15 05:55

【摘要】：目的旨在研究p38分裂原激活的蛋白激酶(MAPK)信号通路和转录因子Osterix在间断性机械牵张力刺激下促进小鼠骨髓间充质干细胞(BMSC)成骨向分化的作用机制。方法将C57BL/6J小鼠BMSC分为空白对照组、牵张力组和牵张力阻断组(p38MAPK通路抑制剂SB203580+牵张力),采用Flexercell体外细胞力学加载装置,施加频率为0.5 Hz、形变率为0.8%的牵张力,每天2次,每次30 min,分别在实验第1、3、5 d收获BMSC。实时荧光定量聚合酶链反应(RT-PCR)检测成骨基因ALP、COL I、OCN的m RNA水平变化情况,Western blot检测P-p38-MAPK蛋白的表达情况。通过小干扰核糖核酸(si RNA)技术沉默小鼠BMSC的osterix基因,Western blot检测Osterix蛋白的表达情况,RT-PCR检测成骨相关基因ALP、COL I、OCN的mRNA水平变化情况。结果牵张力作用后,可观察到成骨相关基因ALP、COL I、OCN及转录因子Osterix的m RNA水平增高。沉默osterix后,小鼠BMSC成骨相关基因ALP、COL I、OCN的m RNA水平也随之降低。Western blot结果显示,牵张力组小鼠BMSC中Osterix和P-p38-MAPK的蛋白质水平明显高于对照组(P0.05)。SB203580作用后,小鼠BMSC成骨相关基因ALP、COL I、OCN和osterix的m RNA水平降低。结论间断性牵张力可通过活化p38MAPK-Osterix通路促进小鼠BMSC成骨分化。
[Abstract]:Objective to investigate the mechanism of p38 mitogen-activated protein kinase (MAPK) signaling pathway and transcription factor Osterix in promoting the osteogenic differentiation of mouse bone marrow mesenchymal stem cells (BMSC) induced by intermittent mechanical tension. Methods C57BL/6J mice with BMSC were divided into three groups: the control group, the stretch group and the p38MAPK pathway inhibitor SB203580 group. The Flexercell in vitro cellular mechanical loading device was used to apply the strain with 0.8% deformed rate of 0.5 Hz, twice a day. The BMSC. was harvested at 30 min, on the 1st day of the experiment and 5 days after the experiment. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the change of m RNA level of osteogenic gene ALP,COL. Western blot was used to detect the expression of P-p38-MAPK protein. The expression of Osterix protein in mouse BMSC was detected by (si RNA) technique. The expression of Osterix protein was detected by Western blot. The mRNA level of osteoblast-associated gene ALP,COL Io CN was detected by RT-PCR. Results it was observed that the level of m RNA of osteoblast-associated gene (ALP,COL) and transcription factor (Osterix) was increased after retraction. After silencing osterix, the m RNA level of BMSC osteoblast related gene ALP,COL Io CN was also decreased. The results showed that the BMSC Osterix and P-p38-MAPK protein levels in the stretch group were significantly higher than those in the control group (P0.05). SB203580. The levels of m RNA of BMSC osteoblast associated gene ALP,COL Io CN and osterix were decreased in mice. Conclusion intermittent stretch can promote osteogenic differentiation of BMSC in mice by activating p38MAPK-Osterix pathway.
【作者单位】：青岛大学附属医院口腔颌面外科山东省教育厅口腔重点实验室;青岛大学附属医院儿童口腔科;
【基金】：山东省高校科技计划(J12LK57) 青岛市黄岛区应用研究与公共卫生专项基金(2014-1-84)~~
【分类号】：R329.2

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