Recombinant arginine deiminase as a differential modulator of inducible (iNOS) and endothelial (eNOS) nitric oxide synthetase activity in cultured endothelial cells
Recombinant arginine deiminase as a differential modulator of inducible (iNOS) and endothelial (eNOS) nitric oxide synthetase activity in cultured endothelial cells
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DOI：
10.1016/s0006-2952(03)00555-0
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发表时间：
2003-11-15
影响因子：
5.8
通讯作者：
Shen, WC
中科院分区：
文献类型：
--
作者：
Shen, LJ;Lin, WC;Shen, WC
精氨酸（一氧化氮合成酶的底物）的细胞外水平的调节是缓解某些产生过量一氧化氮（NO）的病理状况的有前景的方法。然而，由于仅优先抑制诱导型一氧化氮合成酶 (iNOS)，而不是内皮型一氧化氮合成酶 (eNOS)，因此需要优先抑制 NO 过量产生，因此会出现并发症。我们研究了重组精氨酸脱亚胺酶 (rADI) 介导的精氨酸剥夺对内皮细胞系 TR-BBB 中 iNOS 和 eNOS 活性的影响。我们的结果表明，细胞因子诱导的 NO 产生依赖于细胞外精氨酸作为底物。然而，如果培养基中存在足够的瓜氨酸，则 eNOS 产生的 A23187 激活的 NO 不依赖于细胞外精氨酸。 rADI治疗可以显着抑制细胞因子诱导的通过iNOS产生的NO，但不能抑制A23187通过eNOS激活的NO产生。我们的结果还表明，即使在上调 iNOS 表达的条件下，也可以通过消耗培养基中的精氨酸来减少 iNOS 产生的 NO。因此，rADI 似乎是一种新型的 iNOS 活性选择性调节剂，可用作研究 NO 过量产生发挥关键作用的病理性疾病的工具。 (C) 2003 Elsevier Inc. 保留所有权利。
Modulation of the extracellular level of arginine, substrate for nitric oxide synthetases, is a promising modality to alleviate certain pathological conditions where excess nitric oxide (NO) is produced. However, complications arise, as only preferential inhibition of the inducible nitric oxide synthetase (iNOS), but not endothelial nitric oxide synthetase (eNOS), is desired for the treatment of NO overproduction. We investigated the effect of arginine deprivation mediated by a recombinant arginine deiminase (rADI) on the activity of iNOS and eNOS in an endothelial cell line, TR-BBB. Our results demonstrated that cytokine-induced NO production depends on the extracellular arginine as substrate. However, if sufficient citrulline is present in the medium, A23187-activated NO production by eNOS does not rely on extracellular arginine. Treatment with rADI can markedly inhibit cytokine-induced NO production via iNOS, but not A23187-activated NO production via eNOS. Our results also showed that the decrease of NO production by iNOS could be achieved by depleting arginine from the medium even under the conditions that would up-regulate iNOS expression. Thus, rADI appears to be a novel selective modulator of iNOS activity that may be a used as a tool in the study of pathological disorders where NO over-production plays a key role. (C) 2003 Elsevier Inc. All rights reserved.