Egr-1通过上调NDRG1诱导骨髓间充质干细胞成骨分化

发布时间：2018-02-14 21:08

本文关键词： 骨质疏松骨髓间充质干细胞成骨细胞早期生长因子 N-myc下游调节基因　出处：《重庆医学》2017年04期 　论文类型：期刊论文

【摘要】：目的探究早期生长因子1(Egr-1)对骨髓间充质干细胞(BMSCs)增殖和成骨分化的影响。方法采集成年男性骨髓组织,分离培养原代BMSCs并在显微镜下观察其形态,流式细胞术鉴定BMSCs表面标志物;pcDNA3.1/Egr-1转染BMSCs,MTT检测BMSCs的增殖,茜素红钙染色试剂盒检测细胞基质内钙结节的形成,ALP活性测定试剂盒检测细胞内ALP的活性,实时定量qPCR和Western blot分别检测细胞内Egr-1、Runx2、NDRG1mRNA和蛋白表达;Egr-1siRNA转染BMSCs,检测细胞内ALP活性、Egr-1、Runx2和NDRG1mRNA及蛋白表达。结果离体培养的BMSCs高表达CD90和CD29,而CD34和CD45呈阴性表达;pcDNA3.1/Egr-1转染对BMSCs增殖无明显作用,却能促进细胞基质内钙结节的形成,上调ALP活性和Egr-1、Runx2、NDRG1的表达,而Egr-1siRNA则作用相反。结论 Egr-1通过上调NDRG1诱导BMSCs的成骨分化。
[Abstract]:Objective to investigate the effect of early growth factor (Egr-1) on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). BMSCs surface marker pcDNA3.1 / Egr-1 was transfected into BMSCs by flow cytometry to detect the proliferation of BMSCs, and alizarin red calcium staining kit was used to detect the formation of intracellular calcium nodules and the activity of ALP in cells. Real-time quantitative qPCR and Western blot were used to detect the mRNA and protein expression of Egr-1rx2ndRG1mRNA and protein in BMSCs, and the activity of ALP in BMSCs was detected. Results BMSCs in vitro overexpressed CD90 and CD29, while CD34 and CD45 showed negative expression of pcDNA3.1 / Egr-1 on BMSCs. Proliferation has no obvious effect, However, it can promote the formation of calcium nodules in the cell matrix and up-regulate the activity of ALP and the expression of Egr-1 Runx2, NDRG1, but Egr-1siRNA has the opposite effect. Conclusion Egr-1 can induce the osteogenic differentiation of BMSCs by up-regulating NDRG1.
【作者单位】：河南中医药大学第一附属医院内分泌科;河南中医药大学第三附属医院内分泌科;
【分类号】：R580

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