【摘要】：目的旨在研究p38分裂原激活的蛋白激酶(MAPK)信號通路和轉(zhuǎn)錄因子Osterix在間斷性機(jī)械牽張力刺激下促進(jìn)小鼠骨髓間充質(zhì)干細(xì)胞(BMSC)成骨向分化的作用機(jī)制。方法將C57BL/6J小鼠BMSC分為空白對照組、牽張力組和牽張力阻斷組(p38MAPK通路抑制劑SB203580+牽張力),采用Flexercell體外細(xì)胞力學(xué)加載裝置,施加頻率為0.5 Hz、形變率為0.8%的牽張力,每天2次,每次30 min,分別在實(shí)驗(yàn)第1、3、5 d收獲BMSC。實(shí)時熒光定量聚合酶鏈反應(yīng)(RT-PCR)檢測成骨基因ALP、COL I、OCN的m RNA水平變化情況,Western blot檢測P-p38-MAPK蛋白的表達(dá)情況。通過小干擾核糖核酸(si RNA)技術(shù)沉默小鼠BMSC的osterix基因,Western blot檢測Osterix蛋白的表達(dá)情況,RT-PCR檢測成骨相關(guān)基因ALP、COL I、OCN的mRNA水平變化情況。結(jié)果牽張力作用后,可觀察到成骨相關(guān)基因ALP、COL I、OCN及轉(zhuǎn)錄因子Osterix的m RNA水平增高。沉默osterix后,小鼠BMSC成骨相關(guān)基因ALP、COL I、OCN的m RNA水平也隨之降低。Western blot結(jié)果顯示,牽張力組小鼠BMSC中Osterix和P-p38-MAPK的蛋白質(zhì)水平明顯高于對照組(P0.05)。SB203580作用后,小鼠BMSC成骨相關(guān)基因ALP、COL I、OCN和osterix的m RNA水平降低。結(jié)論間斷性牽張力可通過活化p38MAPK-Osterix通路促進(jìn)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.
【作者單位】： 青島大學(xué)附屬醫(yī)院口腔頜面外科山東省教育廳口腔重點(diǎn)實(shí)驗(yàn)室;青島大學(xué)附屬醫(yī)院兒童口腔科;
【基金】：山東省高�？萍加媱�(J12LK57) 青島市黃島區(qū)應(yīng)用研究與公共衛(wèi)生專項基金(2014-1-84)~~
【分類號】：R329.2

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