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首页> 外文期刊>Cell and Tissue Research >Activation of toll-like receptor signaling in endothelial progenitor cells dictates angiogenic potential: from hypothesis to actual state
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Activation of toll-like receptor signaling in endothelial progenitor cells dictates angiogenic potential: from hypothesis to actual state

机译：内皮祖细胞中的Toll样受体信号传导的激活决定了血管生成潜力：从假说到实际状态

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Human endothelial progenitor cells (EPCs) were isolated from cord blood samples and enriched by magnetic activated cell sorting method based on the CD133 marker. Cells were incubated with different doses of bacterial lipopolysaccharide, ranging from 2, 5, 10, 50, 100, 200, 250, 500, to 1000 mu g/ml, for 48 h. The cell survival rate was determined by using MTT assay. To confirm activation of the toll-like receptor signaling pathway, PCR array analysis was performed. Protein levels of ERK1/2, p-ERK1/2, NF-kappa B and TRIF proteins were measured using western blotting. The content of TNF-alpha and lipoprotein lipase activity were analyzed by immunofluorescence imaging. Flow cytometric analysis of CD31 was performed to assess the maturation rate. Cell migration was studied by the Transwell migration assay. The expression of genes related to exosome biogenesis was measured using real-time PCR analysis. In vivo gel plug angiogenesis assay was done in nude mice. Lipopolysaccharide changed endothelial progenitor cells' survival in a dose-dependent manner with maximum viable cells in groups treated with 2 mu g/ml. PCR array analysis showed the activation of toll-like signaling pathways after exposure to LPS (p<0.05). Western blotting analysis indicated an induction of p-ERK1/2 and Erk1/2, NF-kappa B and TRIF in LPS-treated EPCs compared with the control (p<0.05). Immunofluorescence staining showed an elevation of TNF-alpha and lipoprotein lipase activity after lipopolysaccharide treatment (p<0.05). Lipopolysaccharide increased EPC migration and expression of exosome biogenesis-related genes (p<0.05). In vivo gel plug analysis revealed enhanced angiogenesis in cells exposed to bacterial lipopolysaccharide. Data highlighted the close relationship between the toll-like receptor signaling pathway and functional activity in EPCs.
机译：从脐血样本中分离人内皮祖细胞（EPCs），并通过基于CD133标记物的磁激活细胞分选法富集。将细胞与不同剂量的细菌脂多糖（2,5,10,50,100,200,250,500至1000μg/ml）孵育48小时。通过MTT法测定细胞存活率。为了证实toll样受体信号通路的激活，进行了PCR阵列分析。用western印迹法检测ERK1/2、p-ERK1/2、NF-κB和TRIF蛋白的蛋白水平。免疫荧光成像分析TNF-α含量和脂蛋白脂肪酶活性。对CD31进行流式细胞术分析以评估成熟率。细胞迁移通过Transwell迁移试验进行研究。使用实时PCR分析测量与外体生物发生相关的基因表达。在裸鼠体内进行凝胶塞血管生成试验。脂多糖以剂量依赖性的方式改变内皮祖细胞的存活率，在2μg/ml处理组中，最大活细胞数。PCR阵列分析显示，暴露于LPS后，toll样信号通路激活（p<0.05）。Western blotting分析表明，与对照组相比，LPS处理的内皮祖细胞中p-ERK1/2、ERK1/2、NF-κB和TRIF均被诱导（p<0.05）。免疫荧光染色显示脂多糖治疗后TNF-α和脂蛋白脂肪酶活性升高（p<0.05）。脂多糖增加了EPC迁移和外体生物发生相关基因的表达（p<0.05）。体内凝胶塞分析显示，暴露于细菌脂多糖的细胞血管生成增强。数据强调了toll样受体信号通路与内皮祖细胞功能活性之间的密切关系。

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