Nitric oxide suppression of cellular proliferation depends on cationic amino acid transporter activity in cytokine-stimulated pulmonary endothelial cells.
Nitric oxide suppression of cellular proliferation depends on cationic amino acid transporter activity in cytokine-stimulated pulmonary endothelial cells.
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一氧化氮对细胞增殖的抑制取决于细胞因子刺激的肺内皮细胞中的阳离子氨基酸转运蛋白活性。
DOI：
10.1152/ajplung.00029.2010
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Nelin,LeifD
中科院分区：
文献类型：
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作者：
Chicoine,LouisG;Chicione,LouisG;Stenger,MichaelR;Cui,Hongmei;Calvert,Andrea;Evans,RebeccaJ;English,BKeith;Liu,Yusen;Nelin,LeifD
诱导型一氧化氮合酶（Inducible nitric oxide synthase，iNOS）是一种在炎症状态下上调的应激反应蛋白，NO可抑制细胞增殖。我们假设，阻止内皮细胞摄取l-精氨酸（l-arg）可以阻止脂多糖/肿瘤坏死因子-α（LPS/TNF）诱导的、NO介导的细胞增殖抑制。牛肺动脉内皮细胞（bPAEC）用LPS/TNF或媒介物（对照）和10 mM-亮氨酸[1-leu;阳离子氨基酸转运蛋白（CAT）的fl-arg摄取的竞争性抑制剂]或其媒介物处理。在平行实验中，使用腺病毒载体（分别为AdiNOS或AdArgII）在bPAEC中过表达iNOS或iNOS酶II。LPS/TNF处理增加了bPAEC中iNOS、CAT-1和CAT-2 mRNA的表达，导致比对照bPAEC更大的NO和尿素产生，这被yl-leu阻止。LPS/TNF处理导致活细胞比对照组少，而用l-leu处理的LPS/TNF刺激的bPAEC比单独LPS/TNF处理的活细胞更多。LPS/TNF处理导致裂解的caspase-3和裂解的聚（ADP-核糖）聚合酶表达，其被yl-leu减弱。AdiNOS降低了活细胞数，并且用l-leu处理AdiNOS转染的bPAEC保持了细胞数。AdArgII增加了活细胞数，并且用l-leu处理AdArgII转染的bPAEC阻止了细胞数的增加。这些数据表明，肺内皮细胞中的iNOS表达导致细胞增殖降低，这可以通过阻止cellularl-arg摄取来减弱。我们推测，CAT活性可能是一种新的治疗靶点，在炎症性肺部疾病的特点是NO的过度生产。
Inducible nitric oxide (NO) synthase (iNOS) is a stress response protein upregulated in inflammatory conditions, and NO may suppress cellular proliferation. We hypothesized that preventingl-arginine (l-arg) uptake in endothelial cells would prevent lipopolysaccharide/tumor necrosis factor-α (LPS/TNF)-induced, NO-mediated suppression of cellular proliferation. Bovine pulmonary arterial endothelial cells (bPAEC) were treated with LPS/TNF or vehicle (control), and either 10 mMl-leucine [l-leu; a competitive inhibitor ofl-arg uptake by the cationic amino acid transporter (CAT)] or its vehicle. In parallel experiments, iNOS or arginase II were overexpressed in bPAEC using an adenoviral vector (AdiNOS or AdArgII, respectively). LPS/TNF treatment increased the expression of iNOS, arginase II, CAT-1, and CAT-2 mRNA in bPAEC, resulting in greater NO and urea production than in control bPAEC, which was prevented byl-leu. LPS/TNF treatment resulted in fewer viable cells than in controls, and LPS/TNF-stimulated bPAEC treated withl-leu had more viable cells than LPS/TNF treatment alone. LPS/TNF treatment resulted in cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase expression, which was attenuated byl-leu. AdiNOS reduced viable cell number, and treatment of AdiNOS transfected bPAEC withl-leu preserved cell number. AdArgII increased viable cell number, and treatment of AdArgII transfected bPAEC withl-leu prevented the increase in cell number. These data demonstrate that iNOS expression in pulmonary endothelial cells leads to decreased cellular proliferation, which can be attenuated by preventing cellularl-arg uptake. We speculate that CAT activity may represent a novel therapeutic target in inflammatory lung diseases characterized by NO overproduction.