Razumovitch Julia A, Semenkova Galina N, Fuchs Dietmar, Cherenkevich Sergey N
Department of Biophysics, Physics Faculty, Belarus State University, Scoryny avenue 4, 220050 Minsk, Belarus.
FEBS Lett. 2003 Aug 14;549(1-3):83-6. doi: 10.1016/s0014-5793(03)00796-8.
Neopterin is synthesized by human monocyte-derived macrophages primarily upon stimulation with the cytokine interferon-gamma. We studied the influence of neopterin on the generation of reactive oxygen species (ROS) in human peripheral blood neutrophils. Radical formation was measured using a biochemiluminometer. Neutrophils were isolated from peripheral blood of healthy donors. The generation of ROS by neutrophils suspended in Earl's solution (pH=7.4) at 37 degrees C was investigated by monitoring of chemiluminescence using luminol and lucigenin as light emitters. Neopterin induced chemiluminescence in suspensions of neutrophils in the presence of luminol, but not of lucigenin. Neopterin affected only adhesive cells. Addition of neopterin into the suspension of the cells involving D-mannitol, L-histidine and diazabicyclo[2.2.2]octane (DABCO) decreased luminol-dependent chemiluminescence (LDCL) of the neutrophils. The action of superoxide dismutase (SOD) and 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) reduced neopterin-induced LDCL of neutrophils. Data suggest that neutrophils respond on exposure to neopterin with additional generation of singlet oxygen, hydroxyl radical and nitric oxide by nicotinamide adenine dinucleotide phosphate (NADPH)-independent pathways.
新蝶呤主要由人单核细胞衍生的巨噬细胞在细胞因子干扰素-γ刺激下合成。我们研究了新蝶呤对人外周血中性粒细胞中活性氧（ROS）生成的影响。使用生物化学发光计测量自由基的形成。从健康供体的外周血中分离中性粒细胞。通过使用鲁米诺和光泽精作为发光体监测化学发光，研究了在37℃下悬浮于厄尔溶液（pH = 7.4）中的中性粒细胞产生ROS的情况。新蝶呤在鲁米诺存在的情况下诱导中性粒细胞悬液中的化学发光，但在光泽精存在时则不然。新蝶呤仅影响黏附细胞。向含有D-甘露醇、L-组氨酸和二氮杂双环[2.2.2]辛烷（DABCO）的细胞悬液中添加新蝶呤会降低中性粒细胞的鲁米诺依赖性化学发光（LDCL）。超氧化物歧化酶（SOD）和2-苯基-4,4,5,5-四甲基咪唑啉-1-氧基-3-氧化物（PTIO）的作用降低了新蝶呤诱导的中性粒细胞LDCL。数据表明，中性粒细胞在暴露于新蝶呤时会通过烟酰胺腺嘌呤二核苷酸磷酸（NADPH）非依赖性途径额外产生单线态氧、羟基自由基和一氧化氮。
