Lipopolysaccha ride and TNF-α produce very similar changes in gene expression in human endothelial cells
Lipopolysaccha ride and TNF-α produce very similar changes in gene expression in human endothelial cells
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DOI：
10.1159/000095162
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发表时间：
2006-01-01
影响因子：
1.7
通讯作者：
Sladek, R.
中科院分区：
文献类型：
--
作者：
Magder, S.;Neculcea, I.;Sladek, R.
由Toll样受体4（TLR 4）和肿瘤坏死因子-α TNF-α调节的细胞内信号传导途径都激活NF κ B B。这表明脂多糖（LIPS）和TNF-α应该改变一组共同基因的转录。我们通过用LIPS（50 ng/ml + 1 μ g/ml CD 14）或TNF-α（10 ng/ml）处理第一代人脐内皮细胞（HUVEC）6小时并通过微阵列分析（AffyphidaseGeneChips）分析基因表达的变化来检验这一假设。LIPS和TNF-α增加了191个共同基因的表达，降低了102个基因的表达。编码大量趋化因子、粘附分子、促凝血因子和影响细胞完整性的分子的调节转录物。基于微阵列分析和随后通过北方分析对特定基因的确认，所有被LIPS改变的203个基因都被TNF-α改变。另外17个基因仅由TNF-α诱导，46个基因的表达降低。然而，在变化的动力学方面存在一些差异。我们还发现，内源性CD 14是存在于这些早期传代细胞和外源性CD 14是不必要的大部分LIPS反应。通过用单克隆抗体阻断TNF-α，也排除了LPS诱导的TNF-α表达的自分泌效应。总之，LIPS诱导HUVEC中基因表达的强烈改变，这与TNF-α诱导的非常相似。LIPS对内皮细胞的这种作用可能在天然免疫反应中起重要作用。版权所有（c）2006 S. Karger AG，巴塞尔。
Intracellular signaling pathways regulated by Toll-like receptor 4 (TLR4) and tumor necrosis factor-alpha TNF-alpha both activate NF kappa B. This suggests that lipopolysaccharicle (LIPS) and TNF-alpha should alter transcription of a common set of genes. We tested this hypothesis by treating first passage human umbilical endothelial cells (HUVEC) for 6 h with LIPS (50 ng/ml + 1 mu g/ml CD14) or TNF-alpha (10 ng/ml) and analyzing changes in gene expression by microarray analysis (Affymetrix GeneChips). LIPS and TNF-alpha increased expression of 191 common genes and decreased expression of 102 genes. Regulated transcripts encoded for a large number of chemokines, adhesion molecules, procoagulant factors, and molecules that affect cell integrity. Based on the microarray analysis and subsequent confirmation of specific genes by Northern analysis, all 203 genes altered by LIPS were altered by TNF-alpha. An additional 17 genes were induced only by TNF-alpha and the expression of 46 was reduced. There were, however, some differences in the kinetics of changes. We also showed that endogenous CD14 was present on these early passage cells and exogenous CD14 was not necessary for most of the LIPS response. An autocrine effect from LPS induced expression of TNF-alpha also was ruled out by blocking TNF-alpha with monoclonal antibodies. In conclusion, LIPS induces a robust alteration in gene expression in HUVEC that is very similar to that induced by TNF-alpha. This LIPS effect on endothelium could play an important role in the innate immune response. Copyright (c) 2006 S. Karger AG, Basel.