Nitric oxide modulates interleukin-1β and tumor necrosis factor-α synthesis by alveolar macrophages in pulmonary tuberculosis
Nitric oxide modulates interleukin-1β and tumor necrosis factor-α synthesis by alveolar macrophages in pulmonary tuberculosis
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DOI：
10.1164/ajrccm.161.1.9902113
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发表时间：
2000-01-01
影响因子：
24.7
通讯作者：
Lu, LC
中科院分区：
文献类型：
--
作者：
Kuo, HP;Wang, CH;Lu, LC
肺结核患者肺泡巨噬细胞(AM)分泌的白介素1β(IL-1β)和肿瘤坏死因子-α(TNF-α)在机体抵抗分枝杆菌感染中起重要作用。肺结核患者AM中一氧化氮(NO)的产生增加。我们检测了NO是否与结核患者AM合成(IL)-1β和肿瘤坏死因子-α有关。11例肺结核患者和10例正常人经支气管肺泡灌洗获得纯化的AM，加入或不加入NO抑制剂N-G-单甲基-L-精氨酸(N-G-单甲基-L-精氨酸)培养。采用酶联免疫吸附试验和Northern印迹分析检测IL-1β和TNF-α的释放及其信使RNA(MRNAs)表达。肺结核病患者AM释放的IL-1β和TNF-α明显高于正常人AM。L-NMMA可抑制肺结核患者亚硝酸盐、IL-1β和肿瘤坏死因子-α的产生。结核患者外周血中IL-1β和肿瘤坏死因子-α的基因表达上调，而L-尼莫地平可抑制其表达。用抗核因子-kappaB的p65亚基的单抗进行免疫细胞化学染色，发现核因子-kappaB在结核患者AM中高表达并移位到核内，并被L-NMMA抑制。吡咯烷二硫代氨基甲酸酯抑制核因子-kappaB抑制IL-1β和TNF-α的合成。综上所述，结核患者AM促进NO的产生在放大促炎细胞因子的合成方面起着自身调节作用，可能是通过激活核因子-kappaB。
Interleukin (IL)-1 beta and tumor necrosis factor (TNF)-alpha released from alveolar macrophages (AM) in pulmonary tuberculosis (TB) are important in host defense against mycobacterial infection. Nitric oxide (NO) production is enhanced in AM of TB patients. We examined whether NO was implicated in (IL)-1 beta and TNF-alpha synthesis by AM of TB patients. Purified AM were retrieved by bronchoalveolar lavage from 11 TB patients and 10 normal subjects, and were cultured with or without the NO inhibitor N-G-monomethyl-L-arginine (L-NMMA). The release of IL-1 beta and TNF-alpha, and expression of their messenger RNAs (mRNAs), were determined by enzyme-linked immunosorbent assay and Northern blot analysis. The release of IL-1 beta and TNF-alpha was greater from AM of TB patients than from AM of normal subjects. L-NMMA inhibited nitrite, IL-1 beta, and TNF-alpha production in TB patients. The mRNA expression for IL-1 beta and TNF-alpha was upregulated in TB patients and was depressed by L-NMMA. Immunocytochemistry done with a monoclonal antibody against the p65 subunit of nuclear factor (NF)-kappa B showed that NF-kappa B was highly expressed and translocated to the nuclei of AM from TB patients, and was inhibited by L-NMMA. Inhibition of NF-kappa B by pyrrolidine dithiocarbamate attenuated IL-1 beta and TNF-alpha synthesis. In conclusion, enhanced NO generation by AM of TB patients plays an autoregulatory role in amplifying the synthesis of proinflammatory cytokines, probably through NF-kappa B activation.