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-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RT-PCR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。
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-forming 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-forming 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.
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2014年第5期466-470,共5页
Chinese Journal of Osteoporosis
基金
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)
