Kim C D, Kim Y K, Lee S H, Hong K W
Department of Pharmacology, College of Medicine, Pusan National University, Korea.
J Pharmacol Exp Ther. 2000 Sep;294(3):864-9.
This study was designed to determine whether rebamipide can inhibit neutrophil adhesion to human umbilical vein endothelial cells (HUVECs) stimulated with 1 h of hypoxia followed by 4 h of reoxygenation (H/R). Furthermore, to define the action mechanisms, we determined the effect of rebamipide on the surface expression of endothelial cell adhesion molecules E-selectin, P-selectin, and intercellular adhesion molecule-1 (ICAM-1) on H/R-stimulated HUVECs. Under resting conditions, both E-selectin and P-selectin were not expressed on the surface of HUVECs in contrast to ICAM-1, which was constitutively expressed. After stimulation with H/R, HUVECs showed an enhanced neutrophil adhesivity in association with an increased surface expression of E-selectin and P-selectin with a marginal increase in ICAM-1 expression. In parallel, the increased nuclear translocation of nuclear factor-kappaB in H/R-stimulated HUVECs was monitored by electrophoretic mobility shift assay (adjusted volume units, 11.9 +/- 2.5 x 10(4) counts x mm(2) in unstimulated cells versus 24.2 +/- 3.0 x 10(4) counts x mm(2) in H/R-stimulated cells). Rebamipide suppressed the surface expression of E-selectin and P-selectin with a subsequent inhibition of neutrophil adhesion to H/R-stimulated HUVECs. In line with these results, rebamipide (100, 300, and 1000 microM) inhibited H/R-induced nuclear translocation of nuclear factor-kappaB in a concentration-dependent manner. Taken together, this study demonstrates that rebamipide inhibits neutrophil adhesion to HUVECs by a mechanism involving inhibition of transcription-dependent surface expression of E-selectin and P-selectin in H/R-stimulated endothelial cells.
本研究旨在确定瑞巴派特是否能抑制中性粒细胞与经1小时缺氧及4小时复氧（H/R）刺激的人脐静脉内皮细胞（HUVECs）的黏附。此外，为明确作用机制，我们测定了瑞巴派特对H/R刺激的HUVECs上内皮细胞黏附分子E-选择素、P-选择素和细胞间黏附分子-1（ICAM-1）表面表达的影响。在静息条件下，与组成性表达的ICAM-1相反，E-选择素和P-选择素在HUVECs表面均未表达。经H/R刺激后，HUVECs表现出增强的中性粒细胞黏附性，同时E-选择素和P-选择素的表面表达增加，ICAM-1表达略有增加。同时，通过电泳迁移率变动分析监测H/R刺激的HUVECs中核因子-κB核转位增加的情况（未刺激细胞中调整后的体积单位为11.9±2.5×10⁴计数×mm²，H/R刺激细胞中为24.2±3.0×10⁴计数×mm²）。瑞巴派特抑制E-选择素和P-选择素的表面表达，随后抑制中性粒细胞与H/R刺激的HUVECs的黏附。与这些结果一致，瑞巴派特（100、300和1000微摩尔）以浓度依赖的方式抑制H/R诱导的核因子-κB核转位。综上所述，本研究表明瑞巴派特通过抑制H/R刺激的内皮细胞中E-选择素和P-选择素转录依赖性表面表达的机制来抑制中性粒细胞与HUVECs的黏附。
