Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells
Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells
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DOI：
10.1016/s0021-9150(97)00117-2
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发表时间：
1997-11-01
期刊：
影响因子：
5.3
通讯作者：
Shimada, K
中科院分区：
文献类型：
--
作者：
Irokawa, M;Nishinaga, M;Shimada, K
一氧化氮（NO）已被证明可以抑制血小板的粘附和聚集，但没有关于其与凝血系统相互作用的报道。我们研究了l-精氨酸类似物n -硝基- l-精氨酸（LNA）、n - g -硝基- l-精氨酸甲酯（L-NAME）和ng -单甲基- l精氨酸（L-NMMA）这三种竞争性NO生成抑制剂对内皮表面硫酸肝素的影响。在猪主动脉内皮细胞中添加LNA可降低i -125标记的抗凝血酶III与细胞表面硫酸肝素的结合，并呈剂量和时间依赖性。1 mM LNA对抗凝血酶III的抑制作用显著，12 h后10 mM LNA对抗凝血酶III的抑制作用最大，为对照的-50%，L-NAME （1 mM）和L-NMMA （1 mM）也显著抑制抗凝血酶III的结合。铁螯合剂去铁胺明显阻止抗凝血酶III与lnna处理细胞结合的减少。我们进一步研究了L-NAME对内皮细胞细胞内氧化应激的影响，使用氢过氧化敏感荧光染料，羧基二氯-二氢荧光素双乙酸双乙酰氧基甲酯探针，发现L-NAME抑制NO合成导致细胞内氧化应激显著增加。这些结果表明，猪主动脉内皮细胞对NO合成的长期抑制通过增加细胞内氧化应激降低了内皮细胞上抗凝血剂硫酸肝素的表达，这可能是NO影响血管凝血系统的另一种机制。(C) 1997爱思唯尔科学爱尔兰有限公司
Nitric oxide (NO) has been shown to inhibit platelet adhesion and aggregation, but there are no reports on its interaction with the coagulation system. We investigated the effect of the L-arginine analogues, N-nitro-L-arginine (LNA), N-G-nitro-L-arginine methyl ester (L-NAME), and NG-monomethyl-L arginine (L-NMMA), competitive inhibitors of NO production, on endothelial-surface heparan sulfate. Addition of LNA to porcine aortic endothelial cells reduced I-125-labeled antithrombin III binding to the cell surface heparan sulfate in a dose-and time-dependent fashion. Significant inhibition was observed with 1 mM LNA, and the maximal suppression (-50% of control) occurred at 10 mM LNA after 12 h, L-NAME (1 mM) and L-NMMA (1 mM) also significantly inhibited the antithrombin III binding. The iron chelator desferrioxamine significantly prevented the reduction of antithrombin III binding to LNA-treated cells. We further investigated the effect of L-NAME on intracellular oxidative stress of endothelial cells using a hydroperoxide-sensitive fluorochrome, carboxy-dichloro-dihydrofluorescein diacetate bisacetoxymethyl ester probe, and revealed that inhibition of NO synthesis by L-NAME led to a marked increase in intracellular oxidative stress. These results demonstrated that the prolonged inhibition of NO synthesis in porcine aortic endothelial cells decreases the expression of anticoagulant heparan sulfate on endothelial cells through the increase in intracellular oxidative stress, perhaps comprising another mechanism by which NO affects the coagulation system in the vasculature. (C) 1997 Elsevier Science Ireland Ltd.