| 彭程飞1,2 韩雅玲2△ 邓捷2 闫承慧2 康建2 李洋2 李杰1 王艳春3.TNF-α刺激大鼠骨髓间充质干细胞的机制研究[J].,2011,11(7):1201-1204 |
| TNF-α刺激大鼠骨髓间充质干细胞的机制研究 |
| Mechanism of TNF-αin Stimulation of Bone Marrow MesenchymalStem Cells in Rats |
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| DOI： |
| 中文关键词: 骨髓间充质干细胞 肿瘤坏死因子-α 核转录因子-κB 生长因子 |
| 英文关键词: MSCs TNF-α NF-κB the growth factors |
| 基金项目:国家自然科学基金项目(30770793，30971218，81070097)和国家青年科学基金项目（30800465） |
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| 中文摘要: |
| 目的：研究肿瘤坏死因子-α（Tumor necrosis factor-α, TNF-α）刺激大鼠骨髓间充质干细胞（marrow-derived mesenchymal
stem cells, MSCs）的作用机制。方法：采取大鼠骨髓，以密度梯度离心分离出单个核细胞（MNCs），于体外培养并由牛垂体提取物
（PEX）诱导扩增传代培养出骨髓间充质干细胞（MSCs）。经形态学和流式细胞仪检测MSCs 表面标志物鉴定后，用TNF-α 刺激
骨髓间充质干细胞（MSCs），通过酶联免疫吸附剂测定法（enzyme linked immunosorbent assay，ELISA）观察比较不同组别细胞的生
长因子分泌和蛋白印迹法（western blot）来观察细胞中蛋白的变化。结果：①经形态学观察和流式细胞仪检测MSCs 表面标志物鉴
定，提示骨髓间充质干细胞的培养成功。②无TNF-α刺激组与TNF-α刺激组比较，TNF-α刺激组的生长因子分泌显著性增加，
而通过磷酸化IκB 的表达量显著性增加提示NF-κB 被激活(P<0.05)；同时TNF-α 刺激组与TNF-α+NF-κB 抑制剂组比较，
TNF-α+NF-κB 抑制剂组的生长因子分泌显著降低，而通过磷酸化IκB 的表达量显著减少提示NF-κB 的活性被抑制(P<0.05)。
结论：NF-κB 对TNF-α刺激下的骨髓间充质干细胞分泌生长因子有关键性作用。 |
| 英文摘要: |
| Objective: To investigate the promotive effects of NF-κB on the marrow-derived mesenchymal stem cells. Methods:
Mononuclear cells were isolated from rat bone marrow by density-gradient centrifugation and were cultured on fibronectin-coated plates,
supplied with bovine pituitary extract. MSCs were identified by Morphology and Surface molecule markers of MSCs. After the marrow-
derived mesenchymal stem cells were stimulated by TNF-α, the production of the growth factors and the protein expression of
NF-κB in different groups were compared by ELISA and WESTERN BOLT. Results: ① The marrow-derived mesenchymal stem cells
were successful cultured by observing the morphology of the marrow-derived mesenchymal stem cells and detecting the cell surface antigen
by flow cytometry assays. ② The production of the growth factors had significantly increased in TNF-α stimulation group compared
with that of no TNF-α stimulation group. The expression of p-IκB increased significantly indicating the activation of NF-κB (P<0.05).
The production of the growth factors had significantly decreased in stimulation of TNF-α plus the inhibitor of NF-κB group compared
stimulation of TNF-α group, . And the protein expression of p-IκB significantly decressed indicate the inhibition of NF-κB (P<0.05).
Conclusion: NF-κB had crucially protective effects on the marrow-derived mesenchymal stem cells stimulated by TNF-α. |
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