含红花血清干预后大鼠肺动脉内皮细胞Ps、ICAM-1基因表达变化及意义

目的观察含红花血清在缺氧复氧条件下对大鼠肺动脉内皮细胞（RPAECs）P-选择素（Ps）及细胞间黏附分子-1（ICAM-1）基因表达的影响，探讨肺栓塞溶栓后缺血再灌注损伤的机制。方法取SPF级雄性大鼠（18只）RPAECs原代培养后分为四组，常氧组持续通入空气12h；缺氧组通入95％N2、5％CO2混合气12h;复氧组在缺氧1h后通95％O2、5％CO2混合气复氧12h；干预组在缺氧再复氧条件下内皮细胞培养液中加含红花血清2ml。观察不同条件下1、6、12h RPAECs中P5、ICAM-1基因表达的变化。结果常氧组各时间点Ps及ICAM-1 mRNA水平无明显变化，缺氧组各时间点Ps及ICAM-1 mRNA水平有所上调，但与常氧组比较无统计学意义。复氧组各时间点Ps及ICAM-1 mRNA及蛋白表达均明显上调；红花干预组各时间点Ps及ICAM-1 mRNA上调水平明显低于复氧组，P〈0．05。结论缺氧复氧条件下Ps及ICAM-1 mRNA过度表达可能参与了肺栓塞溶栓后缺血再灌注损伤的发病；红花注射液可抑制缺氧复氧条件下Ps及ICAM-1 mRNA的过度表达。     

猪急性心肌梗死再灌注后微血管内皮细胞损伤及通心络的保护作用

目的评价猪急性心肌梗死（AM I）再灌注后微血管内皮细胞结构和功能损伤,以及中药通心络的保护作用和机制。方法 30只中华小型猪随机分成假手术组、模型组、小剂量、中剂量和大剂量通心络治疗组,每组6只。建立AM I再灌注模型,观察不同区域心肌组织内细胞黏附分子和内皮细胞间连接蛋白的含量和变化。结果与正常区比较,模型组再灌注区和无再流区P-选择素（P-selectin）、细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1蛋白及mRNA水平升高,再灌注区内皮钙黏素（VE-cadherin）、无再流区VE-cadherin和β连环蛋白含量降低。与模型组比较,中剂量组再灌注区和无再流区P-selectin和ICAM-1蛋白降低;大剂量组再灌注区P-selectin、ICAM-1蛋白和mRNA含量降低,无再流区P-selectin蛋白和mRNA及ICAM-1蛋白含量降低。大剂量组再灌注区和无再流区VE-cadherin表达升高。结论中药通心络通过抑制细胞黏附分子的表达,减少中性粒细胞浸润和炎症反应,保护AM I再灌注过程内皮细胞屏障功能完整性。     

奇智方对人脐静脉内皮细胞ICAM-1和ICAM-1 mRNA表达的影响

目的采用人脐静脉内皮细胞（HUVEC-304）进行细胞间黏附分子-1（ICAM-1）及其转录水平（ICAM-1mRNA）表达的离体实验研究,观察奇智方对血管内皮功能的作用。方法应用ELISA方法检测ICAM-1;用原位杂交的方法检测ICAM-1 mRNA。结果中药治疗组和阳性对照组与生理盐水组比较,ICAM-1蛋白分子表达均有统计学意义（P〈0．01）;中药治疗组和阳性对照组均能明显使缺氧／复氧后细胞ICAM-1 mRNA下调,与缺氧组比较有统计学意义（P〈0．01）。结论奇智方抑制缺氧／复氧后血管内皮细胞表达ICAM-1和ICAM-1 mRNA,从而保护血管内皮细胞的屏障完整和功能正常,可能是其免疫抑制机理的分子基础之一。     

亚低温对缺氧/复氧内皮细胞胞间黏附分子-1表达和黏附率的影响

目的 观察亚低温对缺氧/复氧内皮细胞胞间黏附分子-1(ICAM-1)表达和黏附率的影响.方法 以ECV304内皮细胞为研究对象,利用简易缺氧室,以缺氧模拟体内缺血性损伤,建立缺氧/复氧内皮细胞模型.将细胞分为33℃及37℃缺氧/复氧组.各组又分为缺氧6,12,18 h后再复氧6 h 3个亚组.应用Western印迹检测两组各亚组ICAM-1的表达并应用细胞计数法检测各亚组细胞和中性粒细胞(PMN)的黏附率.结果 ①在缺氧6,12,18 h亚组,37℃组内皮细胞表达ICAM-1的积分光密度值明显高于33℃组[(0.185±0.026) vs (0.120±0.064)、(0.329±0.057) vs (0.218±0.027)、(0.443±0.068) vs (0.326±0.019)](P均＜0.05).②在缺氧6,12,18 h亚组,33℃组内皮细胞和中性粒细胞的黏附率明显高于37℃组[(29.41±1.77)% vs (22.30±3.25)%、(43.66±2.57)% vs (34.51±5.06)%、(62.92±4.82)% vs (49.68±4.34)%](P＜0.05).结论 亚低温能有效抑制缺氧/复氧内皮细胞与中性粒细胞的黏附,其机制可能与降低缺氧/复氧内皮细胞ICAM-1的表达有关.     

羟基红花黄素A对血管内皮细胞黏附分子的影响

目的考察羟基红花黄素A对氧化损伤血管内皮细胞表达黏附分子[血管内皮细胞黏附分子-1（vascular cell adhesion molecule-1,VCAM-1）和细胞间黏附分子-1（intercellular cell adhesion molecule-1,ICAM-1）]的影响。方法用MTS法测定不同浓度羟基红花黄素A对氧化损伤血管内皮细胞与U937细胞黏附率的影响,用反转录聚合酶链反应（RT-PCR）、酶联免疫吸附试验（ELISA）和Western法检测血管内皮细胞黏附分子的表达。结果羟基红花黄素A可以减轻血管内皮细胞与U937细胞的黏附,反转录聚合酶链反应、酶联免疫吸附试验和Western分析结果表明羟基红花黄素A减轻VCAM-1和ICAM-1的表达。结论羟基红花黄素A可通过减轻黏附分子ICAM-1,VCAM-1的表达,抑制白细胞与血管内皮细胞之间的黏附,从而预防氧化对内皮细胞的损伤。     

二苯乙烯苷对同型半胱氨酸诱导的人脐静脉内皮细胞MCP-1、ICAM-1和VCAM-1表达的影响

目的 观察何首乌二苯乙烯苷对同型半胱氨酸诱导培养的人脐静脉内皮细胞株单核细胞趋化蛋白1(MCP-1)、细胞间黏附分子1(ICAM-1)和血管细胞黏附分子1(VCAM-1) mRNA表达的影响.方法 在人脐静脉内皮细胞的培养基中加入不同浓度的二苯乙烯苷处理2h再加入3.0 mmol/L同型半胱氨酸作用36 h.Hoechst33342核染色检测细胞核损伤;以RT-qPCR检测不同浓度二苯乙烯苷处理对同型半胱氨酸诱导的人脐静脉内皮细胞MCP-1、ICAM-1及VCAM-1 mRNA表达的影响.结果 10 μmol/L浓度以内,二苯乙烯苷预孵育呈浓度依赖性降低3.0 mmol/L同型半胱氨酸所致人脐静脉内皮细胞核损伤加重,抑制同型半胱氨酸所致MCP-1、ICAM-1及VCAM-1 mRNA的表达升高(P＜0.05或P＜0.01).结论 二苯乙烯苷具有明显抑制同型半胱氨酸所致人脐静脉内皮细胞MCP-1、ICAM-1、VCAM-1 mRNA表达增加的作用.     

β-细辛醚对ox-LDL诱导的内皮细胞黏附分子表达的干预作用

目的 通过研究石菖蒲有效成份β-细辛醚对氧化低密度脂蛋白（ox-LDL）诱导的内皮细胞表面黏附分子表达的干预作用，探讨β-细辛醚抗动脉粥样硬化（AS）的作用及其机制。方法 采用流式细胞术和特异性标记单克隆抗体，测定内皮细胞表面细胞间黏附分子-1（ICAM-1，CD54）、血管细胞黏附分子-1（VCAM-1，CD106）、E-选择素（CD62E）和P-选择素（CD62P）的表达率。结果 ox-LDL诱导模型组VCAM-1、ICAM-1、CD62P、CD62E表达明显增高，与正常组比较有显著差异（P〈0.001）,β-细辛醚及维生素C能降低内皮细胞表面黏附分子的表达，与模型组比较有统计意义（P 〈0.05-0.001）。结论 ox-LDL促进内皮细胞表面黏附分子表达，β-细辛醚对ox-LDL有明显的干预作用，能抑制ox-LDL诱导的内皮细胞表面黏附分子表达，有效地保护人内皮细胞免受ox-LDL所致毒性损伤。     

心肌细胞系H9c2缺氧上清诱导人脐静脉内皮细胞ICAM-2的表达

目的探讨心肌细胞H9c2的缺氧培养上清对人脐静脉内皮细胞（HUVEC）细胞间黏附分子-2（ICAM-2）表达的影响及其分子机制。方法大鼠心肌细胞系H9c2在氧浓度为1%的条件下缺氧培养24 h后收集上清,用于HUVEC培养。收集不同时间点HUVEC细胞,提取总RNA和总蛋白,RT-PCR方法检测ICAM-2 mRNA表达,Western杂交方法检测ICAM-2蛋白表达。适当浓度细胞因子孵育HUVEC 6 h,收集细胞提取总蛋白,Western杂交方法检测ICAM-2蛋白表达。ELISA方法测定H9c2缺氧培养上清和常氧培养上清的PDGF-BB和PDGF-AB浓度。结果在mRNA水平,HUVEC在H9c2缺氧培养上清孵育1 h后ICAM-2的表达明显升高,3 h后达到最高;同时在HUVEC表达ICAM-2蛋白在3 h、6 h明显增高。细胞因子刺激实验表明PDGF-BB和PDGF-AB可以使HUVEC表达ICAM-2显著增加。在缺氧上清中PDGF-BB有显著性增加（P〈0.05）,而缺氧上清中的PDGF-AB却有显著减少（P〈0.01）。结论心肌细胞系H9c2的缺氧培养上清可以诱导内皮细胞HUVEC表达ICAM-2,H9c2在缺氧条件下产生的因子PDGF-BB可能是其中起作用的物质。     

ox-LDL对血管内皮细胞促聚集和促黏附相关分子表达的影响

目的　观察血管内皮细胞在不同浓度的氧化型低密度脂蛋白（ox-LDL）诱导及诱导不同时间后内皮细胞促聚集和促黏附活性的动态改变及可能机制。方法　将人脐静脉内皮细胞（HUVEC）株接种在6孔培养板,分别加入不同浓度的ox-LDL（50、100和200　mg/L）并孵育不同时间（12、24和48　h）。放射免疫法测定细胞上清液中前列环素（PGI2）和血栓素A2　（TXA2）含量;Western　blot检测HUVEC中组织因子（TF）、细胞间黏附分子1（ICAM-1）和血管细胞黏附分子1（VCAM-1）的表达。结果　量效关系显示,ox-LDL预处理HUVEC　24　h后,ox-LDL呈浓度依赖性促进HUVEC聚集,表现为PGI2/TXA2比值显著下降（P<0.05或P<0.01）;时效关系显示类似的结果,但以24　h作用最为显著;同时,TF的表达随浓度的升高呈升高趋势。此外,ox-LDL还促进HUVEC黏附,表现为ICAM-1和VCAM-1呈浓度依赖性升高。结论　ox-LDL可增强HUVEC促聚集和促黏附活性,其机制可能与其分泌的PGI2/TXA2下降及上调TF、ICAM-　1和VCAM-　1的表达有关。     

西罗莫司对缺氧/复氧诱导内皮细胞-中性粒细胞黏附的影响及机制

目的　探讨西罗莫司对缺氧 复氧后血管内皮细胞表面黏附分子的表达和中性粒细胞 内皮细胞黏附的影响及机制。方法　采用 β N 乙酰氨基己糖苷酶比色法检测黏附率 ,流式细胞术检测内皮细胞表面黏附分子E 选择素、细胞间黏附分子 1(ICAM 1)的表达 ,Fenton反应测定活性氧 (reactiveoxygenspecies,ROS)的含量 ,Western杂交法检测内皮细胞c JunN端激酶 (JNK)及核因子 κB[nuclearfactor κB ,NF κB(P6 5 ) ]蛋白的表达。结果　血管内皮细胞经缺氧 复氧处理后ROS释放增多 ,JNK及NF κB(P6 5 )蛋白表达增加 ,E 选择素、ICAM 1的表达上调 ,其表面中性粒细胞的黏附增加 ,西罗莫司显著抑制缺氧 复氧的上述作用。结论　西罗莫司抑制缺氧 复氧后血管内皮细胞与中性粒细胞的黏附 ,并可能通过抑制ROS、JNK、NF κB的信号转导途径实现     

Effect of treatment with the antioxidant alpha-lipoic (thioctic) acid on heart and kidney microvasculature in spontaneously hypertensive rats

Endothelial cells represent an important vascular site of signaling and development of damage during ischemia, inflammation and other pathological conditions. Excessive reactive oxygen species production causes pathological activation of endothelium including exposure of cell to adhesion molecules. Intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and platelet-endothelial cell adhesion molecule-1 (PECAM-1) are members of the immunoglobulin super-family which are present on the surface of endothelial cells. These molecules represent important markers of endothelial inflammation. The present study was designed to investigate, with immunochemical and immunohistochemical techniques, the effect of treatment with (+/?)-alpha lipoic (thioctic) acid and its enantiomers on heart and kidney endothelium in spontaneously hypertensive rats (SHR). Arterial hypertension is accompanied by an increased oxidative stress status in the heart characterized by thiobarbituric acid reactive substances (TBARS) and nucleic acid oxidation increase. The higher oxidative stress also modifies adhesion molecules expression. In the heart VCAM-1, which was higher than ICAM-1 and PECAM-1, was increased in SHR. ICAM-1, VCAM-1 and PECAM-1 expression was significantly greater in the renal endothelium of SHR. (+/?)-Alpha lipoic acid and (+)-alpha lipoic acid treatment significantly decreased TBARS levels, the nucleic acid oxidation and prevented adhesion molecules expression in cardiac and renal vascular endothelium. These data suggest that endothelial molecules may be used for studying the mechanisms of vascular injury on target organs of hypertension. The effects observed after treatment with (+)-alpha lipoic acid could open new perspectives for countering heart and kidney microvascular injury which represent a common feature in hypertensive end-organs damage.     

Interferon-gamma and interleukin-4 differentially regulate ICAM-1 and VCAM-1 expression on human lung fibroblasts.

The expression of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and more specifically vascular adhesion molecule-1 (VCAM-1) on lung fibroblasts may be important for migration of inflammatory cells through the submucosa to the airway lumen in the asthmatic inflammatory response. This study aimed to assess which cytokines are regulating ICAM-1 and VCAM-1 expression on human lung fibroblasts. For this purpose, confluent fibroblast cultures (derived from lung tissue from a nonasthmatic donor) were stimulated for 4 h with interleukin (IL)-1beta, tumour necrosis factor (TNF)alpha, interferon (IFN)gamma, IL-4, IL-5 or transforming growth factor (TGF)beta. IL-1beta (optimal concentration (OC) 1 U x mL(-1)) and TNFalpha (OC 100 U x mL(-1)) both increased ICAM-1 and VCAM-1 expression. IFNgamma (OC 2 U x mL(-1)) increased only ICAM-1 expression and IL-4 (OC 5 ng x mL(-1)) increased only VCAM-1 expression, whereas IL-5 (20 ng x mL(-1)) and TGFbeta (10 ng x mL(-1)) did not influence ICAM-1 or VCAM-1 expression. ICAM-1 expression reached a plateau at 8-12 h after cytokine stimulation and remained constant for at least 24 h. VCAM-1 showed a transient increased expression within 24 h after IL-1beta and TNFalpha stimulation. In contrast, VCAM-1 expression did not decrease after maximal expression at 4 h upon IL-4 stimulation. It is concluded that the Helper-1T-cell, type cytokine interferon gamma and the Helper-2 T-cell type cytokine interleukin-4 differentially regulate intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression on human lung fibroblasts. The proinflammatory cytokines interleukin-1beta and tumour necrosis factor alpha increase both intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression, without differential regulation of the expression of these adhesion molecules.     

缺氧—复氧诱导ECV304细胞与中性粒细胞粘附的分子机制

为探讨缺氧-复氧诱导人脐静脉内皮细胞ECV304与中性粒细胞粘附的信号转导机制，以缺氧-复氧诱导粘附为模型，采用比色法检测粘附率，流式细胞术检测ECV304细胞表面粘附分子E选择素、细胞问粘附分子1的表达，Western blot法检测ECV304细胞亲环素A、磷酸化细胞外信号调节激酶、总细胞外信号调节激酶、磷酸化D70核糖体S6激酶、总p70核糖体S6激酶蛋白的表达。结果发现，经缺氧-复氧处理后，ECV304细胞E选择素、细胞间粘附分子1的表达上调，其表面中性粒细胞粘附增加。总细胞外信号调节激酶、总p70核糖体S6激酶蛋白表达无明显改变，亲环素A蛋白表达明显上调，细胞外信号调节激酶和p70核糖体S6激酶显著活化。亲环素A抑制剂环孢霉素A以及亲环素A反义寡核苷酸均明显减轻缺氧-复氧诱导的细胞外信号调节激酶和p70核糖体S6激酶激活，显著减少ECV304细胞与中性粒细胞粘附。p70核糖体S6激酶阻断剂雷帕霉素显著抑制p70核糖体S6激酶的激活，中性粒细胞与ECV304细胞的粘附亦明显减少。细胞外信号调节激酶信号通路特异性阻断剂PD98059亦显著抑制ECV304细胞与中性粒细胞粘附。结果提示，亲环素A-细胞外信号调节激酶-p70核糖体S6激酶信号通路介导缺氧-复氧诱导的ECV304细胞与中性粒细胞粘附。     

Vascular cell adhesion molecule-1 is involved in mediating hypoxia- induced sickle red blood cell adherence to endothelium: potential role in sickle cell disease

We investigated the effects of hypoxia on red blood cell (RBC)- endothelial cell (EC) adherence and the potential mechanism(s) involved in mediating this effect. We report that hypoxia significantly increased sickle RBC adherence to aortic EC when compared with the normoxia controls. However, hypoxia had no effect on the adherence of normal RBCs. In additional studies, we found that the least dense sickle RBCs containing CD36+ and VLA-4+ reticulocytes were involved in hypoxia-induced adherence. We next evaluated the effects of hypoxia on the expression of EC surface receptors involved in RBC adherence to macrovascular ECs, including vascular cell adhesion molecule-1 (VCAM- 1), intracellular adhesion molecule-1 (ICAM-1), and the vitronectin receptor (VnR). Hypoxia upregulated the expression of both VCAM-1 and ICAM-1, whereas no effect on VnR was noted. Potential involvement of VCAM-1 and ICAM-1 in mediating hypoxia-induced sickle RBC-EC adhesion was next investigated using monoclonal antibodies against these receptors. Whereas anti-VCAM-1 had no effect on basal adherence, it inhibited hypoxia-induced sickle RBC adherence in a concentration- dependent manner, with 50% to 75% inhibition noted at 10 to 60 micrograms/mL antibody (n = 6, P < .05 to P < .01). Anti-ICAM-1 (10 to 60 micrograms/mL, n = 8) had no effect on either basal or hypoxia- induced adherence. As noted in the bovine aortic ECs, hypoxia stimulated the adherence of sickle RBCs to human retinal capillary ECs, and this response appeared to be mediated via mechanisms similar to those observed with macro-endothelium, ie, via the adhesive receptor combination VCAM-1-VLA-4. Our studies show that hypoxia enhances sickle RBC adhesion to both macrovascular and human microvascular ECs via the adhesive receptor VCAM-1. Our findings are of interest because hypoxia is an integral part of the pathophysiology of the vaso-occlusive phenomenon in sickle cell anemia.     

Granulocytosis and increased adhesion molecules after resistive loading of the diaphragm.

Upregulation of endothelial cell adhesion molecules, followed by an influx of granulocytes and macrophages, can contribute to exertion-induced skeletal muscle injury. The purpose of this study was to quantify circulating leukocyte subsets, diaphragm injury and infiltrating leukocyte subsets, and surface expression of vascular cell adhesion molecule (VCAM)-1 and intracellular adhesion molecule (ICAM)-1 in the diaphragm after inspiratory resistive loading (IRL). Eight New Zealand white rabbits underwent 1.5 h of IRL and seven control rabbits underwent a sham procedure. Blood samples, taken at baseline and 2, 6, 12, 24, 48 and 72 h after the onset of IRL or sham, showed that band cell counts had increased at 6 h post-IRL. Point counting of haematoxylin and eosin-stained cross-sections, sampled at 72 h post-IRL, showed greater injury in diaphragms from IRL rabbits compared with controls. Immunohistochemical processing showed increased expression of ICAM-1 and VCAM-1, and higher granulocyte and macrophage counts in IRL diaphragms than control diaphragms. Macrophages were the predominant inflammatory cells. Increased intracellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression, and infiltration of granulocytes and macrophages may contribute to inspiratory resistive loading-induced diaphragm injury.     

Perfusion with sickle erythrocytes up-regulates ICAM-1 and VCAM-1 gene expression in cultured human endothelial cells

Sickle cell anemia is characterized by periodic vasoocclusive crises. Increased adhesion of sickle erythrocytes to vascular endothelium is a possible contributing factor to vasoocclusion. This study determined the effect of sickle erythrocyte perfusion at a venous shear stress level (1 dyne/cm(2)) on endothelial cell (EC) monolayers. Sickle erythrocytes up-regulated intercellular adhesion molecule-1 (ICAM-1) gene expression in cultured human endothelial cells. This was accompanied by increased cell surface expression of ICAM-1 and also elevated release of soluble ICAM-1 molecules. Expression of vascular cell adhesion molecule-1 (VCAM-1) messenger RNA (mRNA) was also strikingly elevated in cultured ECs after exposure to sickle cell perfusion, although increases in membrane-bound and soluble VCAM-1 levels were small. The presence of cytokine interleukin-1beta in the perfusion system enhanced the production of ICAM-1 and VCAM-1 mRNA, cell surface expression, and the concentrations of circulating forms. This is the first demonstration that sickle erythrocytes have direct effects on gene regulation in cultured human ECs under well-defined flow environments. The results suggest that perfusion with sickle erythrocytes increases the expression of cell adhesion molecules on ECs and stimulates the release of soluble cell adhesion molecules, which may serve as indicators of injury and/or activation of endothelial cells. The interactions between sickle red blood flow, inflammatory cytokines, and vascular adhesion events may render sickle cell disease patients vulnerable to vasoocclusive crises.     

Immunohistochemical analysis of adhesion molecule expression on muscle biopsy specimens from patients with juvenile dermatomyositis

OBJECTIVE: To assess expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on muscle biopsy specimens from patients with untreated juvenile dermatomyositis (JDM). METHODS: Histochemical and immunohistochemical tests for ICAM-1 and VCAM-1 were performed on serial frozen sections from 27 JDM muscle biopsy specimens. ICAM-1 and VCAM-1 expression was analyzed on capillaries, perimysial and endomysial large vessels, and muscle fibers. Expression was assessed and graded semiquantitatively. RESULTS: Increased ICAM-1 expression was observed on capillaries and perimysial large vessels on semiquantitative analysis, and was statistically more evident than on endomysial large vessels. In all cases, only a few muscle vessels showed expression of VCAM-1. Expression of ICAM-1 and VCAM-1 was observed on few muscle fibers. CONCLUSION: The observation of ICAM-1 expression on muscle vessels, mainly on capillaries of patients with untreated JDM compared to controls, and VCAM-1 expression to a lesser extent, mostly on muscle vessels surrounded by inflammatory infiltrate, supports the participation of these adhesion molecules in the pathologic mechanism of vascular injury in JDM.     

Endothelial nuclear factor-kappaB translocation and vascular cell adhesion molecule-1 induction by complement: inhibition with anti-human C5 therapy or cGMP analogues

We have previously shown that reoxygenation of hypoxic human umbilical vein endothelial cells (HUVECs) leads to the activation and deposition of complement. In the present study, we investigated whether the terminal complement complex (C5b-9) influences HUVEC nuclear factor-kappaB (NF-kappaB) translocation and vascular cell adhesion molecule-1 (VCAM-1) protein expression after hypoxia/reoxygenation by decreasing endothelial cGMP. Additionally, we investigated the action of anti-human C5 therapy on endothelial cGMP, NF-kappaB translocation, and VCAM-1 protein expression. Reoxygenation (0.5 to 3 hours, 21% O(2)) of hypoxic (12 hours, 1% O(2)) HUVECs in human serum (HS) significantly increased C5b-9 deposition, VCAM-1 expression, and NF-kappaB translocation compared with hypoxic/reoxygenated HUVECs treated with the recombinant human C5 inhibitor h5G1.1-scFv. Acetylcholine (ACh)-induced cGMP synthesis was significantly higher in normoxic HUVECs compared with hypoxic HUVECs reoxygenated in HS but did not differ from hypoxic HUVECs reoxygenated in buffer or HS treated with h5G1.1-scFv. Treatment of hypoxic/reoxygenated HUVECs with h5G1.1-scFv or cGMP analogues significantly attenuated NF-kappaB translocation and VCAM-1 protein expression. Treatment with NO analogues, but not a cAMP analogue, cGMP antagonists, or an NO antagonist, also significantly attenuated VCAM-1 expression. We conclude that (1) C5b-9 deposition, NF-kappaB translocation, and VCAM-1 protein expression are increased in hypoxic HUVECs reoxygenated in HS; (2) reoxygenation of hypoxic HUVECs in HS, but not buffer alone, attenuates ACh-induced cGMP synthesis; and (3) treatment of hypoxic/reoxygenated HUVECs with h5G1.1-scFv attenuates C5b-9 deposition, NF-kappaB translocation, and VCAM-1 expression while preserving ACh-induced cGMP synthesis. C5b-9-induced VCAM-1 expression may thus involve an NO/cGMP-regulated NF-kappaB translocation mechanism.