INTRACELLULAR OXIDATIVE STRESS-INDUCED BY NITRIC-OXIDE SYNTHESIS INHIBITION INCREASES ENDOTHELIAL-CELL ADHESION TO NEUTROPHILS
INTRACELLULAR OXIDATIVE STRESS-INDUCED BY NITRIC-OXIDE SYNTHESIS INHIBITION INCREASES ENDOTHELIAL-CELL ADHESION TO NEUTROPHILS
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DOI：
10.1161/01.res.74.6.1133
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发表时间：
1994-06-01
影响因子：
20.1
通讯作者：
KUBES, P
中科院分区：
文献类型：
--
作者：
NIU, XF;SMITH, CW;KUBES, P
本研究的目的是确定是否长期抑制内皮细胞中的一氧化氮合成增加这些细胞的中性粒细胞的表面粘附。将人脐静脉内皮细胞（HUVEC）在48孔微量滴定板中生长至汇合。HUVEC暴露于一氧化氮合成抑制剂NG-硝基-L-精氨酸甲酯（L-NAME）在1小时时不会引起中性粒细胞粘附，但在4小时时以剂量依赖性方式增加粘附。L-精氨酸或一氧化氮供体，但不是cGMP的类似物，防止增加的粘附。针对β（2）-整合素CD 18和内皮细胞粘附分子ICAM-1的单克隆抗体可抑制粘附增加。血小板活化因子（PAF）受体拮抗剂WEB 2086也阻止了L-NAME诱导的中性粒细胞粘附。细胞内氧自由基清除剂（二甲基亚砜，丁基羟基甲苯，和α，α '-联吡啶），铁螯合剂去铁胺，和线粒体抑制剂叠氮化物抑制L-NAME诱导的中性粒细胞粘附，而细胞外氧自由基清除剂（超氧化物歧化酶和过氧化氢酶）没有效果。将HUVEC用2 '，7'-二氯二氢荧光素二乙酸酯加载，并监测氧化成荧光二氯二氢荧光素（DCHF）。荧光增强的L-NAME处理的HUVEC在整个4小时的孵育，一个事件可通过抗氧化剂和叠氮化物。DCHF的细胞内氧化的幅度相当于约0.8 μ mol/L H2 O2。这些数据表明，在HUVECs中，长时间的一氧化氮合成抑制导致中性粒细胞在这些内皮细胞表面上粘附的氧化剂和PAF相关的上升。
The objective of the present study was to determine whether prolonged inhibition of nitric oxide synthesis in endothelial cells increased the surface adhesion of these cells for neutrophils. Human umbilical vein endothelial eels (HUVECs) were grown to confluence in 48-well microtiter plates. Exposure of HUVECs to the nitric oxide synthesis inhibitor NG-nitro-L-arginine methyl ester (L-NAME) did not cause neutrophil adhesion at 1 hour but increased adhesion at 4 hours in a dose-dependent manner. The increased adhesion was prevented with L-arginine or nitric oxide donors but not an analogue of cGMP. The increased adhesion was inhibited by monoclonal antibodies directed against the beta(2)-integrin CD18 and endothelial cell adhesion molecule ICAM-1. Platelet-activating factor (PAF) receptor antagonist WEB 2086 also prevented the L-NAME-induced neutrophil adhesion. Intracellular oxygen radical scavengers (dimethyl sulfoxide, butylated hydroxytoluene, and alpha,alpha'-dipyridyl), the iron chelator desferrioxamine, and the mitochondrial inhibitor azide inhibited the L-NAME-induced neutrophil adhesion, whereas extracellular oxygen radical scavengers (superoxide dismutase and catalase) had no effect. HUVECs were loaded with 2',7'-dichlorodihydrofluorescein diacetate, and oxidation to the fluorescent dichlorodihydrofluorescein (DCHF) was monitored. Fluorescence was enhanced in the L-NAME-treated HUVECs throughout the 4-hour incubation, an event inhibitable by an antioxidant and azide. The magnitude of the intracellular oxidation of DCHF was equivalent to approximate to 0.8 mu mol/L H2O2. These data suggest that prolonged nitric oxide synthesis inhibition in HUVECs causes an oxidant- and PAF-associated rise in adhesion on the surface of these endothelial cells for neutrophils.