An endogenous inhibitor of nitric oxide synthase regulates endothelial adhesiveness for monocytes
An endogenous inhibitor of nitric oxide synthase regulates endothelial adhesiveness for monocytes
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DOI：
10.1016/s0735-1097(00)01013-5
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发表时间：
2000-12-01
影响因子：
24
通讯作者：
Cooke, JP
中科院分区：
文献类型：
--
作者：
Böger, RH;Bode-Böger, SM;Cooke, JP
我们试图确定在培养的人内皮细胞中，不对称二甲基精氨酸（ADMA）是否抑制一氧化氮（NO）的生成，以及这是否与介导单核细胞内皮细胞增殖的氧化剂敏感性信号的激活有关。方法采用功能结合实验研究人脐静脉内皮细胞（ECV 304）和人单核细胞样细胞（THP-1）的功能结合。一氧化氮和超氧阴离子（O-2（-））的测定采用化学发光法，ADMA的测定采用高压液相色谱法，单核细胞趋化蛋白-1（MCP-1）的测定采用ELISA法，NF-κ B的测定采用电迁移凝胶位移法。无生物活性的立体异构体对称二甲基精氨酸不抑制NO的释放。不对称二甲基精氨酸（10 μ M）或天然低密度脂蛋白胆固醇（100 mg/dL）以相同程度增加内皮O-2（-）。不对称二甲基精氨酸也刺激MCP-1形成的内皮细胞。这种效应被氧化还原敏感性转录因子NF-κ B的激活所抵消。内皮细胞与ADMA预孵育后，内皮细胞对THP-1细胞的粘附力呈浓度依赖性增加。不对称二甲基精氨酸诱导的单核细胞结合减少L-精氨酸或中和抗MCP-1 antibody.CONCLUSIONS我们的结论是，内源性NO合成酶抑制剂ADMA在人内皮细胞中合成。不对称二甲基精氨酸增加内皮细胞氧化应激并增强单核细胞结合。不对称二甲基精氨酸可能是一种内源性致动脉粥样硬化分子。(C)2000年，美国心脏病学会。
OBJECTIVES We sought to determine whether asymmetric dimethylarginine (ADMA) inhibits nitric oxide (NO) elaboration in cultured human endothelial cells and whether this is associated with the activation of oxidant-sensitive signaling mediating endothelial adhesiveness for monocytes.BACKGROUND Endothelial NO elaboration is impaired in hypercholesterolemia and atherosclerosis, which may be due to elevated concentrations of ADMA, an endogenous inhibitor of NO synthase.METHODS Human umbilical vein endothelial cells (ECV 304) and human monocytoid cells (THP-1) were studied in a functional binding assay. Nitric oxide and superoxide anion (O-2(-)) were measured by chemiluminescence; ADMA by high pressure liquid chromatography, monocyte chemotactic protein-1 (MCP-1) by:ELISA and NF-KB by electromobility gel shift assay.RESULTS Incubation of endothelial cells with ADMA (0.1 muM to 100 muM) inhibited NO formation, which was reversed by coincubation with L-arginine (1 mM). The biologically inactive stereoisomer symmetric dimethylarginine did not inhibit NO release. Asymmetric dimethylarginine (10 muM) or native low-density lipoprotein cholesterol (100 mg/dL) increased endothelial O-2(-) to the same degree. Asymmetric dimethylarginine also stimulated MCP-1 formation by endothelial cells. This effect was paralleled by activation of the redox-sensitive transcription factor NF-kappaB. Preincubation of endothelial cells with ADMA increased the adhesiveness of endothelial cells for THP-1 cells in a concentration-dependent manner. Asymmetric dimethylarginine-induced monocyte binding was diminished by L-arginine or by a neutralizing anti-MCP-1 antibody.CONCLUSIONS We concluded that the endogenous NO synthase inhibitor ADMA is synthesized in human endothelial cells. Asymmetric dimethylarginine increases endothelial oxidative stress and potentiates monocyte binding. Asymmetric dimethylarginine may be an endogenous proatherogenic molecule. (C) 2000 by the American College of Cardiology.