本文選題：Jagged + 破骨細(xì)胞��； 參考：《中國(guó)免疫學(xué)雜志》2014年07期

【摘要】：目的:探討Jagged1通過(guò)活化Notch通路對(duì)RAW 264.7細(xì)胞向破骨分化及增殖的影響。方法:將RAW 264.7細(xì)胞分三組培養(yǎng):對(duì)照組(細(xì)胞培養(yǎng)基+鼠RANKL 50 ng/L)、Jagged1組(細(xì)胞培養(yǎng)基+鼠RANKL+Jagged1重組蛋白)、γ-分泌酶抑制劑(DAPT)組(細(xì)胞培養(yǎng)基+鼠RANKL+Jagged1重組蛋白+DAPT),實(shí)時(shí)熒光定量PCR檢測(cè)各組破骨細(xì)胞標(biāo)志基因組織蛋白酶K(Cathepsin K,CK)、抗酒石酸性磷酸酶(Tartrate-resistant acid phosphatase,TRAP)、降鈣素受體(Calcitionin receptor,CTR)及Notch靶基因HES-1、HEY-1 mRNA的表達(dá),TRAP染色鑒定破骨細(xì)胞,掃描電鏡檢測(cè)破骨細(xì)胞溶骨功能。免疫熒光檢測(cè)NICD的表達(dá)變化,CCK-8檢測(cè)RAW 264.7細(xì)胞增殖。結(jié)果:Jagged1組TRAP、CK、CTR及HES-1、HEY-1 mRNA的表達(dá)、TRAP+細(xì)胞數(shù)較對(duì)照組及DAPT組明顯升高(P0.05),而DAPT組與對(duì)照組均無(wú)明顯變化;細(xì)胞免疫熒光顯示Jagged1組NICD除表達(dá)于細(xì)胞膜、細(xì)胞質(zhì)外,細(xì)胞核也有較高表達(dá),對(duì)照組及DAPT組細(xì)胞核中NICD無(wú)明顯表達(dá);細(xì)胞培養(yǎng)48 h,Jagged1組細(xì)增殖較對(duì)照組及DAPT組出現(xiàn)明顯抑制。結(jié)論:Jagged1通過(guò)活化Notch通路促進(jìn)RAW 264.7向破骨細(xì)胞分化、抑制其增殖。
[Abstract]:Aim: to investigate the effect of JaggeD1 on osteoclast differentiation and proliferation of raw 264.7 cells by activating Notch pathway. Methods: raw 264.7 cells were cultured in three groups: the control group (RANKL 50 ng / L) and the 緯 -secretory enzyme inhibitor group (RANKL Jagged1 recombinant protein) and the 緯 -secretory enzyme inhibitor group (RANKL Jagged1 recombinant protein DAPT). The expression of cathepsin K (CK), tartrate-resistant acid phosphatase trap (trap), calcitonin receptor (CTR) and Notch target gene HES-1hHEY-1 mRNA in osteoclasts were detected by fluorescence quantitative polymerase chain reaction (FQ-PCR). The osteolytic function of osteoclasts was examined by scanning electron microscope. The expression of NICD was detected by immunofluorescence and CCK-8 was used to detect the proliferation of raw 264.7 cells. Results compared with control group and DAPT group, the number of TRAPCKCTR and HES-1pHEY-1 mRNA expression of TRAPCKCTR and HES-1HEY-1 mRNA in group 1 were significantly higher than those in control group and DAPT group (P0.05), but there were no significant changes in DAPT group and control group, and cell immunofluorescence showed that NICD in Jagged1 group was expressed not only in cell membrane, but also in cytoplasm. The expression of NICD in the nuclei of control group and DAPT group was not obvious, and the fine proliferation of JaggeD1 group was significantly inhibited than that of control group and DAPT group. Conclusion by activating Notch pathway, Jagged1 promotes the differentiation of raw 264.7 into osteoclasts and inhibits its proliferation.
【作者單位】： 第三軍醫(yī)大學(xué)第二附屬醫(yī)院骨科;
【基金】：國(guó)家自然科學(xué)基金面上項(xiàng)目(No.81271979)
【分類號(hào)】：R392

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