Mishra A, Dayal N, Beck-Speier I
GSF-Forschungszentrum für Umwelt und Gesundheit, Projekt Inhalation, Neuherberg, Oberschleissheim, Germany.
J Biolumin Chemilumin. 1995 Jan-Feb;10(1):9-19. doi: 10.1002/bio.1170100103.
To assess the effect of sulphite on the oxidative metabolism of human neutrophils, chemiluminescence (CL) measurements were performed using lucigenin and luminol as chemiluminigenic probes. Lucigenin-dependent CL was used for measuring superoxide anion (O2-) production, and luminol-dependent CL was used for determination of myeloperoxidase (MPO)-connected processes. With sulphite concentrations of 0.01 to 1 mmol/L, resting neutrophils showed an up to sixfold increase of lucigenin-dependent CL, but only a 1.9-fold increase of luminol-dependent CL. Subsequent stimulation of sulphite-treated neutrophils with phorbol myristate acetate (PMA) (soluble stimulant) or zymosan (particulate stimulant) resulted in an additional significant increase of lucigenin-dependent CL compared to stimulated control cells, whereas luminol-dependent CL increased slightly by 0.01 mmol/L sulphite and decreased then continuously. Sulphite concentrations above 1 mmol/L decreased both lucigenin- and luminol-dependent CL of resting and PMA- or zymosan-stimulated neutrophils. Lucigenin-dependent CL of sulphite-treated and subsequently stimulated neutrophils was strongly inhibited by extracellularly added superoxide dismutase, whereas luminol-dependent CL was markedly reduced by the MPO inhibitor azide. The intracellular activity of MPO in neutrophils stimulated with PMA in the presence of sulphite (2 mmol/L) was reduced by 55%. Sulphite (0.1 mmol/L) also inhibited strongly the activity of MPO in a cell-free system. These results indicate that micromolar concentrations of sulphite exert a stimulating effect on the O2- production of neutrophils extracellularly, but have an inhibitory effect on MPO-catalysed reactions intracellularly.
为评估亚硫酸盐对人中性粒细胞氧化代谢的影响，使用光泽精和鲁米诺作为化学发光探针进行化学发光（CL）测量。依赖光泽精的CL用于测量超氧阴离子（O2-）的产生，依赖鲁米诺的CL用于测定髓过氧化物酶（MPO）相关过程。当亚硫酸盐浓度为0.01至1 mmol/L时，静息中性粒细胞依赖光泽精的CL最多增加了6倍，但依赖鲁米诺的CL仅增加了1.9倍。随后用佛波酯肉豆蔻酸酯（PMA）（可溶性刺激剂）或酵母聚糖（颗粒性刺激剂）刺激经亚硫酸盐处理的中性粒细胞，与受刺激的对照细胞相比，依赖光泽精的CL出现了额外的显著增加，而依赖鲁米诺的CL在亚硫酸盐浓度为0.01 mmol/L时略有增加，随后持续下降。高于1 mmol/L的亚硫酸盐浓度降低了静息以及经PMA或酵母聚糖刺激的中性粒细胞依赖光泽精和鲁米诺的CL。细胞外添加超氧化物歧化酶可强烈抑制经亚硫酸盐处理并随后受刺激的中性粒细胞依赖光泽精的CL，而MPO抑制剂叠氮化物可显著降低依赖鲁米诺的CL。在亚硫酸盐（2 mmol/L）存在的情况下，用PMA刺激的中性粒细胞中MPO的细胞内活性降低了55%。亚硫酸盐（0.1 mmol/L）在无细胞系统中也强烈抑制MPO的活性。这些结果表明，微摩尔浓度的亚硫酸盐在细胞外对中性粒细胞的O2-产生具有刺激作用，但在细胞内对MPO催化的反应具有抑制作用。
