Beck-Speier I, Lenz A G, Godleski J J
GSF-Forschungszentrum für Umwelt und Gesundheit, Neuherberg, Germany.
J Toxicol Environ Health. 1994 Mar;41(3):285-97. doi: 10.1080/15287399409531844.
Exposure to sulfur dioxide or sulfite aerosols induce inflammatory reactions in the respiratory tract characterized by an influx of neutrophils into the airways. To determine direct intracellular effects of sulfite on human neutrophils, these cells were evaluated ultrastructurally by electron microscopy and analyzed for their extracellular and intracellular respiratory burst activity after incubation with sulfite (0.01-10 mM) in vitro. The respiratory burst was quantitated by measuring both the extracellular release of superoxide anions (O2-) by superoxide dismutase-inhibitable lucigenin-dependent chemiluminescence (CL) and the intracellular generation of hydrogen peroxide (H2O2) by flow cytometry using the reagent dichlorofluorescein diacetate. The addition of sulfite in concentrations of 0.01-1 mM resulted in sixfold increases in CL of resting neutrophils. Neutrophils stimulated with zymosan, phorbol myristate acetate (PMA), or N-formyl-methionine-leucine-phenylalanine further increased CL when sulfite was added. Higher sulfite concentrations (2-10 mM) decreased CL of resting, zymosan-stimulated, and PMA-stimulated cells. When sulfate was added, no changes in CL of resting and zymosan-stimulated neutrophils were seen, indicating that the effect is specific for sulfite. The intracellular generation of H2O2 in resting and PMA-stimulated neutrophils incubated with sulfite (0.1-2 mM) was increased twofold. These findings suggest that sulfite in low concentrations stimulates neutrophils by activating the respiratory burst to produce O2- and H2O2. Ultrastructural studies confirm the stimulating effect of sulfite on neutrophils with sulfite-treated cells exhibiting increased ruffled surface membranes, degranulation changes, and vesiculation similar to those seen in PMA-stimulated cells.
接触二氧化硫或亚硫酸盐气溶体会在呼吸道引发炎症反应，其特征是中性粒细胞流入气道。为了确定亚硫酸盐对人中性粒细胞的直接细胞内作用，通过电子显微镜对这些细胞进行超微结构评估，并在体外与亚硫酸盐（0.01 - 10 mM）孵育后分析其细胞外和细胞内呼吸爆发活性。通过超氧化物歧化酶抑制的基于光泽精的化学发光（CL）测量超氧阴离子（O2-）的细胞外释放以及使用试剂二氯荧光素二乙酸通过流式细胞术测量过氧化氢（H2O2）的细胞内生成来定量呼吸爆发。添加浓度为0.01 - 1 mM的亚硫酸盐会使静息中性粒细胞的CL增加六倍。用酵母聚糖、佛波酯肉豆蔻酸酯乙酸酯（PMA）或N-甲酰甲硫氨酸-亮氨酸-苯丙氨酸刺激的中性粒细胞在添加亚硫酸盐时CL进一步增加。较高浓度的亚硫酸盐（2 - 10 mM）会降低静息、酵母聚糖刺激和PMA刺激细胞的CL。添加硫酸盐时，静息和酵母聚糖刺激的中性粒细胞的CL未见变化，表明该作用对亚硫酸盐具有特异性。与亚硫酸盐（0.1 - 2 mM）孵育的静息和PMA刺激的中性粒细胞中H2O2的细胞内生成增加了两倍。这些发现表明，低浓度的亚硫酸盐通过激活呼吸爆发刺激中性粒细胞产生O2-和H2O2。超微结构研究证实了亚硫酸盐对中性粒细胞的刺激作用，经亚硫酸盐处理的细胞表现出与PMA刺激细胞类似的增加的皱襞表面膜、脱颗粒变化和囊泡形成。
