【摘要】：目的探討Notch信號抑制對核因子κB受體活化因子配體(RANKL)誘導(dǎo)的小鼠RAW264.7細(xì)胞向破骨細(xì)胞分化的影響。方法建立RANKL誘導(dǎo)的小鼠RAW264.7細(xì)胞體外分化模型,實時定量聚合酶鏈反應(yīng)(real-time PCR)檢測Notch信號成分(Notch1、Notch2、Delta1、Jagged1)、下游靶基因Hes1以及破骨細(xì)胞標(biāo)志基因抗酒石酸酸性磷酸酶(TRAP)和Cathepsin K在誘導(dǎo)前后mRNA的表達(dá)。在誘導(dǎo)體系中加入不同濃度的γ分泌酶抑制劑(GSI),抑制Notch受體的表達(dá),TRAP染色檢測破骨細(xì)胞分化的變化情況。結(jié)果 50 ng·m L-1 RANKL誘導(dǎo)小鼠RAW264.7細(xì)胞3 d,Notch1、Notch2、Delta1、Jagged1及Hes1的mRNA表達(dá)均有不同程度的提高,其中以Notch2、Jagged1增高最明顯;破骨細(xì)胞標(biāo)志基因表達(dá)顯著增高。在RANKL誘導(dǎo)的同時加入不同濃度GSI,抑制Notch的表達(dá),可致Notch下游靶基因Hes1表達(dá)下降,同時TRAP陽性細(xì)胞計數(shù)顯著減少,且呈劑量依賴性。結(jié)論 Notch信號可促進(jìn)RANKL誘導(dǎo)的RAW264.7細(xì)胞向破骨細(xì)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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