Notch信号促进核因子κB受体活化因子配体诱导的破骨细胞分化的体外研究

发布时间：2019-06-07 08:01

【摘要】：目的探讨Notch信号抑制对核因子κB受体活化因子配体(RANKL)诱导的小鼠RAW264.7细胞向破骨细胞分化的影响。方法建立RANKL诱导的小鼠RAW264.7细胞体外分化模型,实时定量聚合酶链反应(real-time PCR)检测Notch信号成分(Notch1、Notch2、Delta1、Jagged1)、下游靶基因Hes1以及破骨细胞标志基因抗酒石酸酸性磷酸酶(TRAP)和Cathepsin K在诱导前后mRNA的表达。在诱导体系中加入不同浓度的γ分泌酶抑制剂(GSI),抑制Notch受体的表达,TRAP染色检测破骨细胞分化的变化情况。结果 50 ng·m L-1 RANKL诱导小鼠RAW264.7细胞3 d,Notch1、Notch2、Delta1、Jagged1及Hes1的mRNA表达均有不同程度的提高,其中以Notch2、Jagged1增高最明显;破骨细胞标志基因表达显著增高。在RANKL诱导的同时加入不同浓度GSI,抑制Notch的表达,可致Notch下游靶基因Hes1表达下降,同时TRAP阳性细胞计数显著减少,且呈剂量依赖性。结论 Notch信号可促进RANKL诱导的RAW264.7细胞向破骨细胞分化。
[Abstract]:Objective to investigate the effect of Notch signal inhibition on the differentiation of mouse RAW264.7 cells into osteoclasts induced by nuclear factor kappa B receptor activating factor ligand (RANKL). Methods the differentiation model of mouse RAW264.7 cells induced by RANKL in vitro was established. Notch signal components (Notch1,Notch2,Delta1,Jagged1) were detected by real-time quantitative polymerase chain reaction (real-time PCR). Expression of downstream target genes Hes1 and osteoclast marker genes resistant to tartrate acid phosphatase (TRAP) and Cathepsin K before and after induction. Different concentrations of gamma secretase inhibitor (GSI), were added to the induction system to inhibit the expression of Notch receptor. The differentiation of osteoclasts was detected by TRAP staining. Results the mRNA expression of RAW264.7 cells induced by 50 ng 路mL 鈮，

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