Human monocyte colony-stimulating factor stimulates the gene expression of monocyte chemotactic protein-1 and increases the adhesion of monocytes to endothelial monolayers.
Human monocyte colony-stimulating factor stimulates the gene expression of monocyte chemotactic protein-1 and increases the adhesion of monocytes to endothelial monolayers.
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DOI：
10.1172/jci116762
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发表时间：
1993-10
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影响因子：
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通讯作者：
Y. Shyy;L. Wickham;J. Hagan;H. Hsieh;Y. Hu;S. Telian;A. Valente;K. Sung;S. Chien
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文献类型：
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作者：
Y. Shyy;L. Wickham;J. Hagan;H. Hsieh;Y. Hu;S. Telian;A. Valente;K. Sung;S. Chien
用重组人单核细胞源性集落刺激因子（MCSF）刺激人脐静脉内皮细胞（HUVEC），可增加单核细胞趋化蛋白（MCP-1）的基因表达。北方印迹分析表明，50 U/ml的MCSF是这种作用的最佳浓度。MCP-1 mRNA的表达在刺激后1 h开始升高，并维持至少8 h。MCSF处理的HUVEC中增加的MCP-1水平也通过使用多克隆MCP-1特异性抗体的免疫细胞化学染色在蛋白质水平上证明。HUVEC经50 U/ml MCSF激活5 h后，免疫荧光染色较对照组明显增强。从HUVEC中微量移液器分离THP-1单核细胞显示，MCSF激活THP-1和内皮细胞导致分离力增加3倍以上（36.2 +/- 6.7 x 10（-4）vs. 9.6 +/- 3.6 x 10（-4）dyn）。MCSF激活外周血单核细胞和HUVEC后，也观察到dyn增加（16.7 +/- 2.7 x 10（-4）vs. 5.2 +/- 0.9 x 10（-4）dyn）。两种系统中增加的粘附力通过使用抗MCP-1（5.5 +/- 0.8 x 10（-4）dyn和6.8 +/- 1.1 x 10（-4）dyn）被阻断。在只有HUVEC而不是单核细胞被MCSF激活的实验中获得了类似的结果。这种未处理的单核细胞与MCSF激活的HUVEC的粘附增加也被添加抗MCP-1阻断。相比之下，仅用MCSF处理THP-1或外周血单核细胞而不处理HUVEC的实验未显示这些细胞之间的粘附显著增加。这些结果表明，MCSF主要通过其对内皮细胞的作用来增强单核细胞与内皮细胞的相互作用，并且该功能可能是通过增加MCP-1的表达来介导的。MCSF/MCP-1依赖的粘附机制可能在体内动脉壁中起作用，导致在动脉粥样硬化形成过程中浸润的单核-巨噬细胞被困在内皮下间隙中。
The stimulation of the human umbilical vein endothelial cell (HUVEC) with recombinant human monocyte-derived colony-stimulating factor (MCSF) increased the gene expression of monocyte chemotactic protein (MCP-1). Northern blot analysis indicated that 50 U/ml of MCSF is the optimal concentration for this effect. The elevation of MCP-1 mRNA started as early as 1 h after stimulation and was maintained for at least 8 h. An increased MCP-1 level in MCSF-treated HUVEC was also demonstrated at the protein level by immunocytochemical staining using a polyclonal MCP-1-specific antibody. HUVEC activated by 50 U/ml of MCSF for 5 h showed a stronger immunofluorescence staining than control cells. Micropipette separation of THP-1 monocytes from HUVEC showed that the activation of both THP-1 and endothelium by MCSF led to an increase in the separation force by more than three times (36.2 +/- 6.7 x 10(-4) vs. 9.6 +/- 3.6 x 10(-4) dyn). An increased adhesiveness was also observed after MCSF activation of peripheral blood monocytes and HUVEC (16.7 +/- 2.7 x 10(-4) vs. 5.2 +/- 0.9 x 10(-4) dyn). The increased adhesive force in both systems was blocked by the use of anti-MCP-1 (5.5 +/- 0.8 x 10(-4) and 6.8 +/- 1.1 x 10(-4) dyn). Similar results were obtained in experiments in which only HUVEC, but not monocytes, were activated by MCSF. This increased adhesion of untreated monocytes to MCSF-activated HUVEC was also blocked by the addition of anti-MCP-1. In contrast, experiments in which only THP-1 or peripheral blood monocytes, but not HUVEC, were treated with MCSF did not show a significant increase of adhesion between these cells. These results indicate that MCSF augments monocyte-endothelium interaction primarily by its action on the endothelial cell and that this function is probably mediated through an increased expression of MCP-1. The MCSF/MCP-1-dependent adhesive mechanism might be operative in the arterial wall in vivo to lead to the trapping of the infiltrated monocyte-macrophage in the subendothelial space during atherogenesis.