Role of leukocyte-endothelial interaction for pathogenesis of vessel diseases perturbed by cytokines

为了研究白细胞-内皮细胞相互作用在细胞因子干扰的血管疾病发病机制中的作用，已经进行了体外实验。即使在基因调控下存在细胞因子的情况下，血流动力学也可调节各种内皮细胞功能。采用改良的锥板式粘度计，在细胞因子或脂多糖（LPS）的作用下，研究了剪切力对培养的人脐静脉内皮细胞（HUVECs）凝血和纤溶系统的影响。已经开发了几种针对人鼻粘膜微血管的单克隆抗体。Eselectin和Mel-CAM是炎症部位白细胞粘附于内皮细胞的重要分子，针对这两种粘附糖蛋白的单克隆抗体已被制备。E-选择素在静息状态下未受刺激的HUVECs中未被检测到，而E-选择素在静息状态下的HUVECs表面膜表达增加。 ...更多信息 TNF刺激后3 ~ 6小时，相比之下，在静息的未刺激的HUVEC中清楚地观察到Mel-CAM表达，而在TNF刺激后，向培养物上清的释放增加，在切应力下减少。我们还研究了ICAM-1和VCAM-1在HUVECs中的表达，采用Western印迹法。TNF刺激后3 ~ 6 h，ICAM-1和VCAM-1的表达明显增加。我们已经从人外周血中分离出单个核细胞（MNCs），使用Ficoll梯度法。未观察到MNC与静息未刺激的HUVEC的粘附，而在TNF刺激后6 ~ 9小时清楚地观察到白细胞与HUVEC的粘附。有趣的是，抑制NF-κB的活化可降低这些细胞粘附分子的表达和白细胞对TNF刺激的HUVECs的粘附。这些结果表明，在TNF刺激下，通过NF-κB活化调节白细胞-内皮细胞相互作用可能是通过细胞粘附分子调节炎症和动脉粥样硬化的重要调节剂。少

To investigate the role of leukocyte-endothelial interaction for pathogenesis of vessel diseases perturbed by cytokines, in vitro experiments have been undertaken. Hemodynamic forces modulate various endothelial cell functions even in the presence of cytokines under gene regulation. We have investigated the effect of shear stress on coagulation and fibrinolysis system in cultured human umbilical vein endothelial cells (HUVECs) perturbed by cytokines or lipopolysaccharide (LPS), using modified cone-plate type viscometer, in which well controlled and defined shear forces were generated. Several monoclonal antibodies have been developed against human nasal-mucous microvessels. Monoclonal antibodies have been produced against two adhesive glycoproteins for Eselectin and Mel-CAM that act as important molecules for leucocyte adhesion to endothelium under inflammatory site. E-selectin was not detected in resting unstimulated HUVECs, whereas, surface membrane expression of E-selectin was incre … More ased 3〜6 hours after the stimulation of TNF. In contrast, Mel-CAM expression was clearly observed in resting unstimulated HUVECs, whereas the release to culture supernatant was increased after TNF stimulation and decreased under shear stress. We also investigated the expression of ICAM-1 and VCAM-1 in HUVECs, using western blotting. ICAM-1 and VCAM-1 was not detected in resting unstimulated HUVECs, whereas, the expression of ICAM-1 and VCAM-1 have been increased 3〜6 hours after the stimulation of TNF. We have isolated mononuclear cells (MNCs) from human peripheral blood, using Ficoll-gradient methods. No adhesion of MNCs was observed to resting unstimulated HUVECs, whereas, the leukocyte adhesion to HUVECs was clearly observed 6〜9 hours after TNF-stimulation. Interestingly the expression of these cytoadhesion molecules and leukocyte adhesion to TNF-stimulated HUVECs have been decreased by the inhibition of NF-κB activation. These results indicate that the regulation of leukocyte-endothelial interaction under TNF-stimulation through NF-κB activation might be very important modulator for inflammation and atheroclerosis through cytoadhesion molecules. Less