Effects of ambient air particles on nitric oxide production in macrophage cell lines
Effects of ambient air particles on nitric oxide production in macrophage cell lines
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DOI：
10.1023/b:cbto.0000038461.02222.95
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发表时间：
2004-07-01
影响因子：
6.1
通讯作者：
Vincent, R
中科院分区：
文献类型：
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作者：
Chauhan, V;Breznan, D;Vincent, R
我们评估了不同颗粒对三种小鼠巨噬细胞系（J774A.1, WR19M）的体外毒性。1、RAW264.7)。将细胞暴露于含有城市颗粒物（SRM-1648、SRM-1649、EHC-93）、细颗粒物（PM2.5）、二氧化钛（SRM-154b）和可呼吸性石英石（SRM-1879）的水悬浮液（0-100杯/30毫米(2)孔）中2小时，然后用脂多糖（LPS, 100 ng/ml）和重组干扰素γ (IFN, 100 U/ml)刺激细胞。与颗粒和LPS/IFN孵育过夜后，从培养上清亚硝酸盐中估计一氧化氮的产量。通过监测AlamarBlue（TM）的还原率来测定细胞活力。亚硝酸盐和活力的剂量效应关系建模为幂函数（Fold change=[Dose+1](beta)），其中beta表示剂量-反应曲线的斜率。效价定义为根据细胞活力校正的亚硝酸盐产量变化率(β （Potency)= β（亚硝酸盐）- β（活力））。总体而言，城市颗粒物降低了一氧化氮的生成（β (POTENCY)0）。二氧化钛（TiO2）基本上是无活性的（beta（POTENCY）接近于0）。Western blot分析发现，暴露于城市颗粒物的细胞中一氧化氮生成的减少与诱导型一氧化氮（iNOS）表达的减少直接相关。结果表明，颗粒是小鼠巨噬细胞一氧化氮产生的调节剂，并可能直接破坏伴随病原体暴露时iNOS的表达。导致NO生成增强导致细胞损伤的途径，以及导致NO释放减少导致细菌清除率降低的途径，都可能与环境颗粒对健康的影响有关。
We assessed the in vitro toxicity of various particles on three murine macrophage cell lines, (J774A.1, WR19M.1, RAW264.7). The cells were exposed to aqueous suspensions (0-100 mug/30 mm(2) well) of urban particulate matter (SRM-1648, SRM-1649, EHC-93), fine particulate matter (PM2.5), titanium dioxide (SRM-154b), and respirable cristobalite (SRM-1879) for 2 h and were then stimulated with lipopolysaccharide (LPS, 100 ng/ml) and recombinant interferon-gamma (IFN, 100 U/ml). After overnight incubation with the particles and LPS/IFN, nitric oxide production was estimated from culture supernatant nitrite. Cell viability was determined by monitoring the rate of AlamarBlue(TM)reduction. The dose-effect relationships for nitrite and viability were modeled as a power function (Fold change=[Dose+1](beta)), where beta represents the slope of the dose-response curve. Potency was defined as the rate of change in nitrite production corrected for cell viability (beta(POTENCY)=beta(NITRITE)-beta(VIABILITY)). Overall, the urban particles decreased nitric oxide production (beta(POTENCY)0). Titanium dioxide (TiO2) was essentially inactive (beta(POTENCY)approximate to 0). The decrease in nitric oxide production seen in cells exposed to the urban particles was directly correlated to a decrease in the expression of inducible nitric oxide (iNOS) as determined by Western blot analysis. The results indicate that particles are modulators of nitric oxide production in murine macrophages and may directly disrupt expression of iNOS during concomitant pathogen exposure. Pathways leading to enhanced NO production causing cell injury, and to decreased NO release resulting in lower bacterial clearance, may both be relevant to the health effects of ambient particles.