Lipopolysaccharide and Sphingosine-1-Phosphate Cooperate To Induce Inflammatory Molecules and Leukocyte Adhesion in Endothelial Cells
Lipopolysaccharide and Sphingosine-1-Phosphate Cooperate To Induce Inflammatory Molecules and Leukocyte Adhesion in Endothelial Cells
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DOI：
10.4049/jimmunol.1201309
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发表时间：
2012-12-01
影响因子：
4.4
通讯作者：
Garcia-Rodriguez, Carmen
中科院分区：
文献类型：
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作者：
Fernandez-Pisonero, Isabel;Duenas, Ana I.;Garcia-Rodriguez, Carmen
鉴于TLRs和鞘氨醇-1-磷酸（S1P）是炎症的关键参与者，我们探索了TLRs和S1P在原代人内皮细胞粘附/炎症通路中的潜在相互作用。Western blot和流式细胞术检测发现，与单独使用任何一种配体相比，LPS （TLR4配体）和S1P处理的细胞明显增强了ICAM-1和E-selectin等粘附分子的表达。观察了e -选择素上调的细胞类型差异。而TLR2/TLR1配体对ICAM-1和e -选择素没有协同作用。与粘附分子表达的增加一致，与单独使用任何一种配体相比，LPS加S1P处理内皮细胞可显著增强剪切应力条件下PBMCs的粘附，并且显示出与tnf - α处理后相当的细胞粘附强度水平。通过ELISA和Western blot检测，LPS和S1P共同增加了促炎分子IL-6、环氧化酶-2、prostacyclin的表达。信号通路分析揭示了LPS + S1P处理HUVEC和人主动脉内皮细胞时ERK的协同磷酸化以及p38和NF-kappa B激活的细胞类型差异。此外，药理学和小干扰RNA实验揭示了S1P（1/3）和NF-kappa B参与了协同效应，细胞起源决定了S1P受体和参与的信号通路。LPS + S1P处理对鞘氨醇激酶活性的诱导表明S1P-鞘氨醇激酶轴参与。综上所述，LPS和S1P共同增加内皮细胞中的促炎分子，进而增强白细胞粘附，从而加剧了S1P介导的促粘/促炎特性。免疫学杂志，2012,19(9):542 - 544。
Given that TLRs and sphingosine-1-phosphate (S1P) are key players in inflammation, we explored the potential interplay between TLRs and S1P in the adhesion/inflammatory pathways in primary human endothelial cells. As determined by Western blot and flow cytometry, cells treated with LPS (a TLR4 ligand) and S1P showed significantly enhanced expression of adhesion molecules such as ICAM-1 and E-selectin compared with the effect of either ligand alone. Cell-type differences on E-selectin upregulation were observed. In contrast, no cooperation effect on ICAM-1 or E-selectin was observed with a TLR2/TLR1 ligand. Consistent with an increase in adhesion molecule expression, endothelial cell treatment with LPS plus S1P significantly enhanced adhesion of PBMCs under shear stress conditions compared with the effect of either ligand alone and exhibited comparable levels of cell adhesion strength as those after TNF-alpha treatment. Moreover, LPS and S1P cooperated to increase the expression of proinflammatory molecules such as IL-6, cyclooxygenase-2, and prostacyclin, as determined by ELISA and Western blot. The analysis of signaling pathways revealed the synergistic phosphorylation of ERK upon LPS plus S1P treatment of HUVEC and human aortic endothelial cells and cell-type differences on p38 and NF-kappa B activation. Moreover, pharmacological and small interfering RNA experiments disclosed the involvement of S1P(1/3) and NF-kappa B in the cooperation effect and that cell origin determines the S1P receptors and signaling routes involved. Sphingosine kinase activity induction upon LPS plus S1P treatment suggests S1P-Sphingosine kinase axis involvement. In summary, LPS and S1P cooperate to increase proinflammatory molecules in endothelial cells and, in turn, to augment leukocyte adhesion, thus exacerbating S1P-mediated proadhesive/proinflammatory properties. The Journal of Immunology, 2012, 189: 5402-5410.