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鲁米诺化学发光与活化中性粒细胞产生活性氧

Luminol chemiluminescence and active oxygen generation by activated neutrophils.

作者信息

Takahashi R, Edashige K, Sato E F, Inoue M, Matsuno T, Utsumi K

机构信息

Department of Medical Biology, Kochi Medical School, Japan.

出版信息

Arch Biochem Biophys. 1991 Mar;285(2):325-30. doi: 10.1016/0003-9861(91)90367-r.

Abstract

Upon stimulation by various ligands and membrane perturbers, neutrophils produce various active oxygen species. Since luminol chemiluminescence (LCL) in neutrophils can be blocked by azide, an inhibitor of myeloperoxidase, LCL has been believed to reflect mainly the myeloperoxidase-catalyzed reaction. When cells were stimulated by formyl-methionyl-leucyl-phenylalanine, LCL was strongly inhibited by superoxide dismutase (SOD) and uric acid, a scavenger for hydroxy radical (.OH) and singlet oxygen, whereas it was stimulated by azide. LCL was also inhibited by .OH scavengers, such as mannitol, ethanol, and dimethylsulfoxide. However, when stimulated by phorbol myristate acetate or opsonized zymosan, LCL was strongly inhibited by azide but not by uric acid, and the inhibitory action of SOD was low. Thus, the qualitative and quantitative aspects of reactive oxygen generation by activated neutrophils differ significantly from one ligand to another. These results suggest that the metabolic fate of active oxygens in neutrophils and, hence, their effect on microorganisms and the surrounding tissues might differ depending on the stimulus.

摘要

在受到各种配体和膜扰动剂刺激后,中性粒细胞会产生各种活性氧物质。由于中性粒细胞中的鲁米诺化学发光(LCL)可被髓过氧化物酶抑制剂叠氮化物阻断,因此一直认为LCL主要反映髓过氧化物酶催化的反应。当细胞受到甲酰甲硫氨酰亮氨酰苯丙氨酸刺激时,LCL受到超氧化物歧化酶(SOD)和尿酸(一种羟基自由基(·OH)和单线态氧清除剂)的强烈抑制,而叠氮化物则会刺激LCL。LCL也受到·OH清除剂(如甘露醇、乙醇和二甲基亚砜)的抑制。然而,当受到佛波酯或调理酵母聚糖刺激时,LCL受到叠氮化物的强烈抑制,但不受尿酸抑制,且SOD的抑制作用较弱。因此,活化中性粒细胞产生的活性氧在定性和定量方面因配体不同而有显著差异。这些结果表明,中性粒细胞中活性氧的代谢归宿,进而它们对微生物和周围组织的影响可能因刺激因素而异。

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