文章摘要
杨 姣,夏 雷,汤 郁,陆 化,费小明.肿瘤坏死因子预处理促进骨髓间充质干细胞成骨分化潜能[J].南京医科大学学报,2014,(3):275~280
肿瘤坏死因子预处理促进骨髓间充质干细胞成骨分化潜能
Promotion of tumor necrosis factor-α on osteogenic differentiation of bone marrow derived mesenchymal stem cells
投稿时间:2013-11-29  
DOI:10.7655/NYDXBNS20140301
中文关键词: 骨髓间充质干细胞  肿瘤坏死因子  炎症  NF-κB  成骨分化
英文关键词: bone-marrow mesenchymal stem cells  tumor necrosis factor-α  inflammation  NF-κB  osteogenic differentiation
基金项目:国家自然科学基金(81202358);镇江市社会发展项目(SH2010030,SH2011021)
作者单位
杨 姣 江苏大学附属医院血液科,江苏 镇江 212001 
夏 雷 江苏大学附属医院血液科,江苏 镇江 212001 
汤 郁 江苏大学附属医院风湿科,江苏 镇江 212001 
陆 化 南京医科大学第一附属医院血液科,江苏 南京 210029 
费小明 江苏大学附属医院血液科,江苏 镇江 212001 
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中文摘要:
      目的:观察肿瘤坏死因子(tumor necrosis factor,TNF)-α预处理骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)后,对其核因子(nuclear factor,NF)-κB通路以及成骨分化能力的影响?方法:实验分为BMMSCs阴性对照组(A组)?TNF-α预刺激1 d组(B组)及TNF-α预刺激7 d组(C组)?预刺激结束后,采用Western blot方法检测各组IκBα和p-IκBα蛋白水平的表达;采用RT-PCR方法检测各组BMMSCs中分泌红细胞生成素和肝细胞受体B4(erythroprotein-producing hepatocellular receptor B4,EPHB4)?胰岛素样生长因子1(insulin-like growth factor 1,IGF-1)?骨保持素(osteoprotegerin,OPG)?白细胞介素(interleukin,IL)-7及基质金属蛋白酶(metrix metalloproteinase,MMP)-1 mRNA表达水平的变化;加成骨诱导剂诱导后,茜素红及碱性磷酸酶检测各组成骨分化能力?同时观察加用NF-κB抑制剂吡咯烷二硫氨基甲酸酯(pyrrolidine dithiocarbamate,PDTC)对TNF-α作用的影响?结果:与A组相比,B组和C组IκBα蛋白的表达水平降低(P < 0.05),p-IκBα蛋白的表达水平增高(P < 0.05);与A组相比,B组和C组的EPHB4?IGF-1?OPG mRNA表达水平上调(P < 0.05),IL-7及MMP-1 mRNA表达水平下调(P < 0.05);与A组相比,B组和C组的成骨分化能力增强,而加用NF-κB抑制剂后,B组和C组的成骨分化能力减弱?结论:体外实验表明,BMMSCs经TNF-α预处理后,可以促使BMMSCs成骨分化活性,并且多次刺激较单次刺激效果明显,其促进成骨分化效果至少部分是通过NF-κB通路介导的?
英文摘要:
      Objective:To investigate the effect of continuous pulsed-tumor necrosis factor(TNF-α) treatment on nuclear factor-κB(NF-κB) pathway and osteogentic potentials of bone marrow mesenchymal stem cells (BMMSCs). Methods:There were three groups in this study:Group A:negtive control;Group B:BMMSCs treated by TNF-α once;Group C:BMMSCs continuously pulsed-treated by TNF-α for seven days. After the treatment,IκBα and phospho-IκBα (p-IκBα) were detected by Western blot. The mRNA expression of EPHB4,IGF-1,OPG,IL-7 and MMP-1 in BMMSCs were measured by Real-time PCR. Alizarin red staining and alkaline phosphatase(ALP) staining were employed for the measurement of osteogenic differentiation. This experiment also investigated the effect of the addition of NF-κB inhibitor (pyrrolidine dithiocarbamate,PDTC) on TNF-α action. Results:Compared with group A,IκBα protein levels were lower,while phospho-IκBα(p-IκBα) were higher in both group B and C(P < 0.05). Besides,the mRNA expression levels of EPHB4,IGF-1,OPG were up-regulated but IL-7 and MMP-1 down-regulated after TNF-α priming (P < 0.05). The osteogenic potential of TNF-α-primed BMMSCs was enhanced and the pro-osteogenic differentiation effect of TNF-α was reversed in the presence of NF-κB pathway inhibitor PDTC. Conclusion:Our in vitro results indicated that continuously pulsed TNF-α treatment significantly enhances the osteogenic differentiation of BMMSCs and this effect may be mediated at least partially via NF-κB pathway.
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