Arias M, Zabaleta J, Rodríguez J I, Rojas M, París S C, García L F
Laboratorio Central de Investigaciones, Facultad de Medicina, Universidad de Antioquía, Medellín, Colombia.
Allergol Immunopathol (Madr). 1997 Nov-Dec;25(6):280-8.
Production of nitrix oxide (NO-) by human macrophages is controversial. In the present study, the ability of human monocyte-derived macrophages (M phi) to produce NO- in response to M phi modulators was tested. M phi cultured for up to nine days and stimulated for 48 with different concentrations of LPS and/or IFN-gamma failed to produce significant amounts of NO2- compared to unstimulated cultures. Inhibition of the cyclo-oxygenase pathway with indomethacin did not increase NO2- production by LPS stimulated M phi. Since human M phi lack biopterin, needed for NO- synthesis by murine M phi, human M phi stimulated with LPS plus IFN-gamma were additionally cultured in the presence of neopterin or biopterin. These treatments did not induce NO2- production. Furthermore, simultaneous treatment with indomethacin and neopterin or biopterin also failed to induce NO2- production. However, human M phi, stimulated with IFN-gamma and LPs, produced TNF-alpha suggesting that the lack of increment in NO2- production was not due to an absence of response of M phi to the stimuli used. As an indirect approach to explore the NO- production, human M phi were infected with virulent Mycobacterium tuberculosis H37Rv and simultaneously treated with the competitive inhibitor NGmonomethyl-L-arginine (NGMMA). Mycobacterial intracellular replication was measured by 3H-uracil incorporation. NGMMA did not have any effect on mycobacterial replication. These results further suggest that human M phi do not produce NO- at least by the inducible pathway.
人类巨噬细胞产生一氧化氮（NO-）存在争议。在本研究中，测试了人类单核细胞衍生的巨噬细胞（M phi）对M phi调节剂产生NO-的能力。与未刺激的培养物相比，培养长达九天并用不同浓度的脂多糖（LPS）和/或γ干扰素刺激48小时的M phi未能产生大量的NO2-。用吲哚美辛抑制环氧化酶途径并未增加LPS刺激的M phi产生的NO2-。由于人类M phi缺乏鼠类M phi合成NO-所需的生物蝶呤，因此在用LPS加γ干扰素刺激的人类M phi中额外添加新蝶呤或生物蝶呤进行培养。这些处理并未诱导NO2-的产生。此外，同时用吲哚美辛和新蝶呤或生物蝶呤处理也未能诱导NO2-的产生。然而，用γ干扰素和LPS刺激的人类M phi产生了肿瘤坏死因子-α（TNF-α），这表明NO2-产生缺乏增加并非由于M phi对所用刺激无反应。作为探索NO-产生的间接方法，用强毒力结核分枝杆菌H37Rv感染人类M phi，并同时用竞争性抑制剂N-甲基-L-精氨酸（NGMMA）进行处理。通过3H-尿嘧啶掺入法测量分枝杆菌的细胞内复制。NGMMA对分枝杆菌的复制没有任何影响。这些结果进一步表明，人类M phi至少不会通过诱导途径产生NO-。
