Jagged1活化Notch通路促进RAW 264.7向破骨分化但抑制增殖

发布时间：2018-06-26 20:02

本文选题：Jagged + 破骨细胞　；参考：《中国免疫学杂志》2014年07期

【摘要】：目的:探讨Jagged1通过活化Notch通路对RAW 264.7细胞向破骨分化及增殖的影响。方法:将RAW 264.7细胞分三组培养:对照组(细胞培养基+鼠RANKL 50 ng/L)、Jagged1组(细胞培养基+鼠RANKL+Jagged1重组蛋白)、γ-分泌酶抑制剂(DAPT)组(细胞培养基+鼠RANKL+Jagged1重组蛋白+DAPT),实时荧光定量PCR检测各组破骨细胞标志基因组织蛋白酶K(Cathepsin K,CK)、抗酒石酸性磷酸酶(Tartrate-resistant acid phosphatase,TRAP)、降钙素受体(Calcitionin receptor,CTR)及Notch靶基因HES-1、HEY-1 mRNA的表达,TRAP染色鉴定破骨细胞,扫描电镜检测破骨细胞溶骨功能。免疫荧光检测NICD的表达变化,CCK-8检测RAW 264.7细胞增殖。结果:Jagged1组TRAP、CK、CTR及HES-1、HEY-1 mRNA的表达、TRAP+细胞数较对照组及DAPT组明显升高(P0.05),而DAPT组与对照组均无明显变化;细胞免疫荧光显示Jagged1组NICD除表达于细胞膜、细胞质外,细胞核也有较高表达,对照组及DAPT组细胞核中NICD无明显表达;细胞培养48 h,Jagged1组细增殖较对照组及DAPT组出现明显抑制。结论:Jagged1通过活化Notch通路促进RAW 264.7向破骨细胞分化、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 纬 -secretory enzyme inhibitor group (RANKL Jagged1 recombinant protein) and the 纬 -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.
【作者单位】：第三军医大学第二附属医院骨科;
【基金】：国家自然科学基金面上项目(No.81271979)
【分类号】：R392

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