| 吴颖星,杨卿,杨开祥,等.NF-κB shRNA转染骨髓间充质干细胞对炎症因子的影响.骨科,2014,5(2): 69-72. |
| NF-κB shRNA转染骨髓间充质干细胞对炎症因子的影响 |
| Effect of NF-κB shRNA transfection on expression of inflammatory cytokines in mesenchymal stem cells |
| 投稿时间：2013-12-11 |
| DOI：10.3969/j.issn.1674-8573.2014.02.002 |
| 中文关键词: NF-κB RNA 间质干细胞 骨关节炎 |
| 英文关键词: NF-kappa B RNA Mesenchymal stem cells Osteoarthritis |
| 基金项目:家自然科学基金资助项目（31070831） |
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| 摘要点击次数: 5843 |
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| 中文摘要: |
| 目的 鉴定NF-κB/p65-shRNA质粒并转染入骨髓间充质干细胞，检测靶细胞在NF-κB通路激活时磷酸化NF-κB/p65的表达。方法 用双酶电泳鉴定所得质粒。使用HP试剂将NF-κB/p65-shRNA质粒转染至小鼠BMSCs，在荧光显微镜下观察绿色荧光，并且以流式细胞术检测转染效率，最后以Western Blot检测转染后靶细胞在炎症环境下对磷酸化NF-κB/p65表达的抑制情况。结果 NF-κB/p65-shRNA质粒酶切鉴定结果与预期一致；流式细胞术检测转染效率达到52.76%；Western Blot检测在炎症环境刺激下，转染后靶细胞磷酸化NF-κB/p65表达明显降低。结论 成功将NF-κB/p65-shRNA质粒转染至BMSCs中并有效表达，为进一步实验通过BMSCs修复和重建骨关节炎患者的软骨组织奠定基础。 |
| 英文摘要: |
| Objective To identify the NF-κB/p65 plasmid and to explore the effectiveness of gene silencing in mouse bone marro mesenchymal stem cells (BMSCs). Methods The plasmid was confirmed by restriction analysis and then transfected into BMSCs by HP. Fluorescence microscope and flow cytometry were used to detect the transfection efficiency. Western blotting was performed to determine the protein level of P-NF-κB/p65. ResultsThe result of restriction analysis was consistent with the expected effect. Flow cytometry showed the transfection efficiency was 52.76%. BMSCs expressed a lower level of P-NF-κB/p65 in the inflammatory environment. Conclusion The NF-κB/p65-shRNA plasmid has been successfully transfected into BMSCs and well expressed. It will be a useful method to explore the cartilage restoration and regeneration with BMSCs in osteoarthritis patients. |
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