PYRROLIDINE DITHIOCARBAMATE INHIBITS INDUCTION OF NITRIC-OXIDE SYNTHASE ACTIVITY IN RAT ALVEOLAR MACROPHAGES
PYRROLIDINE DITHIOCARBAMATE INHIBITS INDUCTION OF NITRIC-OXIDE SYNTHASE ACTIVITY IN RAT ALVEOLAR MACROPHAGES
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DOI：
10.1006/bbrc.1993.1359
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发表时间：
1993-03-31
影响因子：
3.1
通讯作者：
IGNARRO, LJ
中科院分区：
文献类型：
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作者：
SHERMAN, MP;AEBERHARD, EE;IGNARRO, LJ
由于核因子κB （NF-κB）在许多炎症条件下被激活，我们评估了它在大鼠肺泡巨噬细胞表达l -精氨酸-一氧化氮途径中的作用。以脂多糖（LPS）和干扰素-γ (ifn -γ)刺激培养的巨噬细胞，加入抑制NF-κB活化的吡咯烷二硫代氨基甲酸酯（PDTC）。通过测定l -精氨酸氧化过程中稳定的氮氧化物终产物亚硝酸盐（NO2−）和硝酸盐（NO3−）来测定诱导型一氧化氮合酶（iNOS）的活性。10、25和50 μM PDTC逐渐抑制巨噬细胞的iNOS活性。在LPS + IFNγ前2 h加入50 μM PDTC，巨噬细胞对l -精氨酸的氧化被>抑制99%；如果同时加入50 μM PDTC和刺激物，l -精氨酸氧化率降低70%；LPS + ifn - γ后6 h，添加50 μM PDTC， NO2−和NO3−未显著降低。环己亚胺与LPS + IFNγ一起添加完全抑制iNOS活性，而在LPS + IFNγ后6 h加入环己亚胺对组织培养上清中的NO2 -和NO3 -没有降低作用。这些结果提示LPS + IFNγ处理巨噬细胞iNOS活性需要NF-κB活化。
Since nuclear factor kappa B (NF-κB) is activated during many inflammatory conditions, we assessed its role in expression of the L-arginine-nitric oxide pathway by rat alveolar macrophages. Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB activation, was added to cultured macrophages stimulated with lipopolysaccharide (LPS) and interferon-γ (IFNγ). Inducible nitric oxide synthase (iNOS) activity was determined by measuring the stable nitrogen oxide end products of L-arginine oxidation: nitrite (NO2−) and nitrate (NO3−). Ten, 25 and 50 μM PDTC progressively inhibited iNOS activity by macrophages. When 50 μM PDTC was added 2 h before LPS + IFNγ, L-arginine oxidation by macrophages was inhibited by >99%; L-arginine oxidation was reduced by 70% if 50 μM PDTC and the stimuli were introduced together; NO2−and NO3−were not decreased significantly if 50 μM PDTC was added 6 h after LPS + IFNγ. Cycloheximide added along with LPS + IFNγ totally inhibits iNOS activity, while cycloheximide added 6 h after LPS + IFNγ did not reduce NO2−and NO3−in tissue culture supernatants. These findings suggest iNOS activity in macrophages treated with LPS + IFNγ requires NF-κB activation.