MACROPHAGE SYNTHESIS OF NITRITE, NITRATE, AND N-NITROSAMINES - PRECURSORS AND ROLE OF THE RESPIRATORY BURST
MACROPHAGE SYNTHESIS OF NITRITE, NITRATE, AND N-NITROSAMINES - PRECURSORS AND ROLE OF THE RESPIRATORY BURST
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DOI：
10.1073/pnas.84.18.6369
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发表时间：
1987-09-01
影响因子：
11.1
通讯作者：
MARLETTA, MA
中科院分区：
文献类型：
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作者：
IYENGAR, R;STUEHR, DJ;MARLETTA, MA
巨噬细胞系RAW 264.7被大肠杆菌脂多糖和干扰素- γ激活后。合成亚硝酸盐（NO2-）和硝酸盐（NO3-）。活化后的培养基变化表明，l -精氨酸是该合成所需的唯一氨基酸。d -精氨酸不能代替l -精氨酸。其他可以取代l-精氨酸的类似物有l-同型精氨酸、l-精氨酸甲酯、l-精氨酸酰胺和l-精氨酸- l-天冬氨酸肽。l -精氨酸、l -谷丙氨酸、l -鸟氨酸、尿素、l -瓜氨酸和氨是非前体，而l -菜豆氨酸抑制这种l -精氨酸衍生的NO2-/NO3-合成。将morpholine加入活化的RAW 264.7巨噬细胞培养基中，发生n -亚硝化反应，生成N-nitrosomorpholine。用L-[胍- 15n2]精氨酸进行的GC/MS实验证实，n -亚硝基somorpholine的NO2-/NO3-和亚硝基完全由精氨酸的一个或两个末端胍氮衍生而来。色谱分析表明，l -精氨酸合成NO2-/NO3-的另一个产物是l -瓜氨酸。采用巨噬细胞系J774.16和J774 C3C研究了呼吸爆发在NO2-/NO3-合成中的作用。两种细胞系合成了相似数量的NO2-/NO3-。然而，J774 C3C细胞不产生超氧化物，因此不表现出呼吸爆发。另外的实验也排除了呼吸爆发参与NO2-/NO3-合成的可能性。
The macrophage cell line RAW 264.7 when activated with Escherichia coli lipopolysaccharide and interferon-.gamma. synthesized nitrite (NO2-) and nitrate (NO3-). Medium change after the activation showed that L-arginine was the only amino acid essential for this synthesis. D-Arginine would not substitute for L-arginine. Other analogues that could replace L-arginine were L-homoarginine, L-arginine methyl ester, L-arginamide, and the peptide L-arginyl-L-aspartate. L-Argininic acid, L-agmatine, L-ornithine, urea, L-citrulline, and ammonia were among the nonprecursors, while L-canavanine inhibited this L-arginine-derived NO2-/NO3- synthesis. When morpholine was added to the culture medium of the activated RAW 264.7 macrophages, N-nitrosation took place, generating N-nitrosomorpholine. GC/MS experiments using L-[guanido-15N2]arginine established that the NO2-/NO3- and the nitrosyl group of N-nitrosomorpholine were derived exclusively from one or both of the terminal guanido nitrogens of arginine. Chromatographic analysis showed that the other product of the L-arginine synthesis of NO2-/NO3- was L-citrulline. The role of the respiratory burst in NO2-/NO3- synthesis was examined using the macrophage cell lines J774.16 and J774 C3C. Both cell lines synthesized similar amounts of NO2-/NO3-. However, J774 C3C cells do not produce superoxide and hence do not exhibit the respiratory burst. Additional experiments also ruled out the involvement of the respiratory burst in NO2-/NO3- synthesis.