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标题：Inhibition of neutrophil superoxide generation by shikonin is associated with suppression of cellular Ca2+ fluxes
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作者：Kimiko Kazumura ; Lucia Satiko Yoshida ; Akiko Hara 等
期刊名称：Journal of Clinical Biochemistry and Nutrition
印刷版ISSN：0912-0009
电子版ISSN：1880-5086
出版年度：2016
卷号：59
期号：1
页码：1-9
DOI：10.3164/jcbn.16-4
出版社：The Society for Free Radical Research Japan
摘要：Shikonin, an anti-inflammatory compound of “Shikon”, inhibits the neutrophil superoxide (O₂^•−) generation by NADPH oxidase 2 (Nox2); however, the mechanisms of how shikonin affects Nox2 activity remained unclear. We aimed to elucidate the relationship between the inhibition of Nox2 activity and influences on intracellular Ca²⁺ concentration ([Ca²⁺]_i) by shikonin. For this purpose, we used a simultaneous monitoring system for detecting changes in [Ca²⁺]_i (by fluorescence) and O₂^•− generation (by chemiluminescence) and evaluated the effects of shikonin on neutrophil-like HL-60 cells stimulated with N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLP). Since fMLP activates Nox2 by elevation in [Ca²⁺]_i via fluxes such as inositol 1,4,5-trisphosphate-induced Ca²⁺ release (IICR) and store-operated Ca²⁺ entry (SOCE), we also evaluated the effects of shikonin on IICR and SOCE. Shikonin dose-dependently inhibited the fMLP-induced elevation in [Ca²⁺]_i and O₂^•− generation (IC₅₀ values of 1.45 and 1.12 µM, respectively) in a synchronized manner. Analyses of specific Ca²⁺ fluxes showed that shikonin inhibits IICR and IICR-linked O₂^•− generation (IC₅₀ values: 0.28 and 0.31 µM for [Ca²⁺]_i and O₂^•−, respectively), as well as SOCE and SOCE-linked O₂^•− generation (IC₅₀ values: 0.39 and 0.25 µM for [Ca²⁺]_i and O₂^•−, respectively). These results suggested that shikonin inhibits the O₂^•− generation by Nox2 in fMLP-stimulated neutrophils by targeting Ca²⁺ fluxes such as IICR and SOCE.
关键词：simultaneous detection;intracellular calcium;superoxide;NADPH oxidase;shikonin