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通过化学发光对兔肺泡巨噬细胞功能进行定量评估。

Quantitative assessment of rabbit alveolar macrophage function by chemiluminescence.

作者信息

Brennan P C, Kirchner F R

出版信息

Environ Res. 1985 Aug;37(2):452-60. doi: 10.1016/0013-9351(85)90127-6.

Abstract

Rabbit alveolar macrophages (RAM) were cultured for 24 hr with concentrations ranging from 3 to 12 micrograms/ml of vanadium oxide (V2O5), a known cytotoxic agent, or with high-molecular-weight organic by-products from coal gasification processes. After culture the cells were harvested and tested for functional capacity using three types of indicators: (1) luminol-amplified chemiluminescence (CL), which quantitatively detects photon emission due to respiratory burst activity measured in a newly designed instrument with standardized reagents; (2) the reduction of nitro blue tetrazolium-saturated polyacrylamide beads, a semiquantitative measure of respiratory burst activity; and (3) phagocytic efficiency, defined as percentage of cells incorporating immunoglobulin-coated polyacrylamide beads. Chemiluminescence declined linearly with increasing concentrations of V2O5 over the dose range tested. Dye reduction and phagocytic efficiency similarly decreased with increasing V2O5 concentration, but were less sensitive indicators of functional impairment than CL as measured by the amount required to reduce the response to 50% of untreated cells. The effect of coal gasification condensates on RAM function varied, but in general these tests also indicated that the CL response was the most sensitive indicator.

摘要

将兔肺泡巨噬细胞(RAM)与浓度范围为3至12微克/毫升的五氧化二钒(V2O5,一种已知的细胞毒性剂)或煤气化过程产生的高分子量有机副产物一起培养24小时。培养后收获细胞,并使用三种类型的指标测试其功能能力:(1)鲁米诺增强化学发光(CL),其通过在配备标准化试剂的新设计仪器中测量呼吸爆发活性来定量检测光子发射;(2)硝基蓝四氮唑饱和聚丙烯酰胺珠的还原,这是呼吸爆发活性的半定量测量;(3)吞噬效率,定义为摄取免疫球蛋白包被的聚丙烯酰胺珠的细胞百分比。在所测试的剂量范围内,化学发光随五氧化二钒浓度的增加呈线性下降。染料还原和吞噬效率同样随五氧化二钒浓度的增加而降低,但与CL相比,它们作为功能损害指标的敏感性较低,CL是通过将反应降低至未处理细胞的50%所需的量来衡量的。煤气化冷凝物对RAM功能的影响各不相同,但总体而言,这些测试也表明CL反应是最敏感的指标。

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