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| 人骨髓间充质干细胞中Notch信号上调对破骨细胞增殖的调控 |
| The effect of up-regulated Notch signaling on the proliferation of osteoclasts by hBMSCs |
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| DOI： |
| 中文关键词: Notch信号；人骨髓间充质干细胞 破骨细胞 |
| 英文关键词:Notch signaling pathway hBMSCs Osteoclast |
| 基金项目:国家重点基础研究发展计划（2011CB964703)，国家高技术研究发展计划（2012AA020502-6) |
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| 中文摘要: |
| 目的通过研究Notch信号上调的人骨髓间充质干细胞（human Bone Marrow Mesenchymal Stem Cells，hBMSCs)对破骨细胞增殖调控的机制，探讨Notch信号在成骨H破骨偶联中的作用。方法用装载Notch信号胞内域NICD1的慢病毒载体(Lentivirus，Lv)转染hBMSCs，收集其分泌的条件培养液(Conditioned Medium，CM)。培养前破骨细胞系RAW264. 7，模拟共培养环境。定量RTTCR和ELISA测定转染前后hBMSCs表达和分泌的M~CSF的情况。cck~8和单板克隆试验测定RAW264 7的增殖情况。结果转染慢病毒后，hBMSCs表达和分泌的M~CSF均有所上调（分别为P <0. 01和P <0. 05)；CCK-8和单板克隆试验的结果显示hBMSCs分泌的CM使RAW264 7的细胞数量增加（均为P < 0. 01)。结论Notch信号上调的hBMSCs可以通过影响M~CSF的表达和分泌，作用于破骨前体细胞，促进其增殖。 |
| 英文摘要: |
| Objective By investigating the mechanism of up-regulated Notch signaling in the proliferation of osteoclasts in human bone marrow mesenchymal stem cells (hBMSCs)，to explore the effect of Notch signaling on the coupling of osteoblasts and osteoclasts. Methods hBMSCs were transfected with the Lentivirus vector (Lv)，which loaded with the Notch intracellular domain 1 (NICD1). And the conditioned medium (CM) was collected. The pre-osteoclast cell line，RAW264. 7，was cultured in the CM，which could mimic the co-culture environment. The expression and secretion of M-CSF in the hBMSCs before and after the transfection were detected using quantitative RT-PCR and ELISA method. The proliferation of RAW264. 7 cells was detected using CCK-8 and colony^orming unit assays. Results After the transfection of Lv，both the expression and secretion of M-CSF in the hBMSCs increased (P <0.01； P <0. 05). Furthermore，the number of RAW 264. 7 cells cultured in the CM secreted by the hBMSCs also increased，according to the detection using CCK-8 and colony■Torming unit assays (P <0. 01). Conclusion The up-regulated Notch signaling can promote the proliferation of the pre-osteoclasts by improving the expression and secretion of M-CSF in the hBMSCs. |
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