| 邓捷韩雅玲彭程飞闫承慧田孝祥康建.体外缺血缺氧条件下大鼠骨髓间充质干细胞凋亡发生的机制研究[J].,2011,11(17):3208-3211 |
| 体外缺血缺氧条件下大鼠骨髓间充质干细胞凋亡发生的机制研究 |
| Hypoxia and Serum Deprivation Induced Bone Mesenchymal StemCells apoptosis Via PI3K/Akt* |
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| DOI： |
| 中文关键词: 骨髓间充质干细胞 缺血缺氧 PI3K/Akt 凋亡 |
| 英文关键词: BMSCs hypoxia and serum deprivation PI3K/Akt apoptosis |
| 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：研究体外大鼠骨髓间充质干细胞（Bone marrow-derived mesenchymal stem cells, BMSCs）在缺血缺氧条件下发生凋亡
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，骨髓间充质干细胞（BMSCs）在缺血
缺氧条件下培养，通过Annexin V/PI 双染细胞凋亡检测比较不同组别细胞的凋亡率和蛋白印迹法（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培养成功。②对照组(无缺血
缺氧) 与缺血缺氧组比较，缺血缺氧组的凋亡率显著性增加，而通过磷酸化Akt 的表达量显著性增加提示PI3K(Phosphoinositide-
3kinase)/Akt(ProteinkinaseB,PKB)信号通路被激活(P<0.05)；同时缺血缺氧组与缺血缺氧+PI3K/Akt 抑制剂(LY294002)组比
较，缺血缺氧+PI3K/Akt 抑制剂(LY294002)组的凋亡率显著降低，而通过磷酸化Akt 的表达量显著减少提示PI3K/Akt 信号通路
被抑制(P<0.05)。结论：PI3K/Akt 信号通路对体外缺血缺氧条件下培养的骨髓间充质干细胞凋亡发生有关键性作用。 |
| 英文摘要: |
| Objective: To investigate the protective effects of PI3K (Phosphoinositide-3kinase)/Akt (ProteinkinaseB,PKB) 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. BMSCs were identified by Morphology and
Surface molecule markers of BMSCs. After the marrow-derived mesenchymal stem cells were under hypoxia and serum deprivation
(hypoxia and SD), the proportion of apoptosis and the protein expression of p-Akt in different groups were compared by Fluorescence-activated
cell sorter (FACS) analysis and WESTERN BOLT. Results: ① The marrow-derived mesenchymal stem cells were successful cultured
by detecting the morphology of the marrow-derived mesenchymal stem cells and the cell surface antigen by using flow cytometry
assays. ② The proportion of apoptosis had significantly increased in the group under hypoxia and SD compared with that of group under
non hypoxia and SD. The expression of p-Akt increased significantly which indicated the activation of PI3K/Akt (P<0.05). The proportion
of apoptosis had significantly decreased in the group under hypoxia and SD plus the inhibitor of PI3K/Akt compared with that in the
group under hypoxia and SD alone. The protein expression of p-Akt significantly decressed, which indicated the inhibition of PI3K/Akt
(P<0.05). Conclusion: PI3K/Akt had crucially protective effects on the marrow-derived mesenchymal stem cells under hypoxia and SD. |
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