Fibrinogen regulates the expression of inflammatory chemokines through NF-κB activation of endothelial cells
Fibrinogen regulates the expression of inflammatory chemokines through NF-κB activation of endothelial cells
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DOI：
10.1160/th04-04-0261
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发表时间：
2004-10-01
影响因子：
6.7
通讯作者：
Francis, CW
中科院分区：
文献类型：
--
作者：
Guo, M;Sahni, SK;Francis, CW
本研究的目的是描述纤维蛋白原通过NF-κ B激活在内皮细胞中刺激炎症趋化因子表达的作用。将人脐静脉内皮细胞（HUVEC）暴露于高达3，000 μ g/ml的纤维蛋白原，并使用电泳迁移率变动测定（EMSA）评估NF-κ B活化。纤维蛋白原暴露导致NF-κ B活化的浓度依赖性增加，4小时后在1，000 μ g/ml时达到最大值，并持续长达24小时。这种作用被α（nu）β（3）和α（5）β（1）抗体和GRGDS肽抑制，表明整合素参与。Mn 2+预孵育降低了纤维蛋白原浓度依赖性，与整合素活化一致。超移位分析表明NF-κ B的p50、p65和c-Rel组分参与。纤维蛋白原暴露也导致单核细胞趋化蛋白-1（MCP-1）和白细胞介素-8的表达上调，如RNA酶保护试验和实时RT-PCR所示。通过ELISA证实MCP-1分泌增加。Parthenidine，一种IkappaB激酶抑制剂，阻止了MCP-1被纤维蛋白原上调，将这种反应与NF-κ B激活联系起来。从我们的研究结果中，我们得出结论，纤维蛋白原调节NF-κ B活化和炎症趋化因子在血管内皮细胞的表达，并可能参与介导炎症过程。
The objective of this study was to characterize the role of fibrinogen in stimulating expression of inflammatory chemokines in endothelial cells through NF-kappaB activation. Human umbilical vein enclothelial cells (HUVEC) were exposed to fibrinogen up to 3,000 mug/ml, and NF-kappaB activation was assessed using electrophoretic mobility shift assay (EMSA). Fibrinogen exposure resulted in a concentration dependent increase in NF-kappaB activation that reached a maximum at 1,000 mug/ml after 4 hours and was sustained up to 24 hours. The effect was inhibited by antibodies to alpha(nu)beta(3) and alpha(5)beta(1) and by the GRGDS peptide, indicating integrin involvement. Preincubation with Mn2+ lowered the fibrinogen concentration-dependence, consistent with integrin activation. Supershift assays demonstrated involvement of the p50, p65 and c-Rel components of NF-kappaB. Fibrinogen exposure also resulted in up-regulation of expression of monocyte chemoattractant protein-1 (MCP-1) and of interleukin-8 as shown by RNase protection assays and by real-time RT-PCR. Increased secretion of MCP-1 was confirmed by ELISA. Parthenolide, an IkappaB kinase inhibitor, prevented up-regulation of MCP-1 by fibrinogen, linking this response to NF-kappaB activation. From our findings, we conclude that fibrinogen regulates NF-kappaB activation and expression of inflammatory chemokines in enclothelial cells and may be involved in mediating inflammatory processes.