Effects of MCI-186 upon neutrophil-derived active oxygens
Effects of MCI-186 upon neutrophil-derived active oxygens
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DOI：
10.1179/135100007x200317
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发表时间：
2007-08-01
期刊：
影响因子：
3.8
通讯作者：
Nakamura, M.
中科院分区：
文献类型：
--
作者：
Sumitomo, K.;Shishido, N.;Nakamura, M.
用分光光度法和质谱法研究了3-甲基-1-苯基-2-吡唑啉-5-酮（MCI-186）与次氯酸和超氧阴离子的反应。将结果应用于嗜中性粒细胞系统，以评价MCI-186对嗜中性粒细胞衍生的活性氧的清除活性。MCI-186与次氯酸（1 × 10（6）M（-1）s（-1））迅速反应形成氯化中间体，然后缓慢转化为新光谱。MCI-186消耗3摩尔次氯酸，不与超氧化物反应。新合成的荧光探针2-[ 6-（4 '-氨基）苯氧基-3H-蒽-3-酮-9-基]苯甲酸（APF）和2-[ 6-（4'-羟基）苯氧基-3H-蒽-3-酮-9-基]苯甲酸（HPF）成功地检测了嗜水气单胞菌衍生的活性氧（Setsukinai K，Urano Y，Kakinuma K，Majima HJ，Nagano T.开发能够可靠检测活性氧并区分特定物种的新型荧光探针。J Biol Chem 2003; 278：3170-3175）。次氯酸与MCI-186和荧光探针反应的速率常数为MCI-186 > APF > HPF。在刺激的中性粒细胞中观察到由于APF和HPF的氧化而产生的荧光。APF氧化的强度高于HPF氧化的强度，这一结果与APF选择性地与次氯酸反应的报道相一致。当反应在荧光探针和MCI-186存在下以剂量依赖性方式进行时，由于APF和HPF的氧化引起的荧光降低。这些结果表明，MCI-186有效清除中性粒细胞来源的次氯酸和其他活性氧。
Reactions of 3-methyl-1-phenyl-2-pyrazoline-5-one ( MCI-186) with hypochlorous acid and superoxide were analysed by spectrophotometry and mass spectrometry. The results were applied to the neutrophil system to evaluate the scavenging activity of neutrophil-derived active oxygen species by MCI-186. MCI-186 reacted rapidly with hypochlorous acid ( 1 x 10(6) M(-1)s(-1)) to form a chlorinated intermediate, followed by a slow conversion to a new spectrum. MCI-186 consumed 3 moles of hypochlorous acid and did not react with superoxide. The newly synthesized fluorescence probes, 2-[ 6-( 4'-amino) phenoxy-3H-xanthen-3-on-9-yl] benzoic acid ( APF) and 2-[ 6-( 4'-hydroxy)phenoxy-3H-anthen-3-on-9-yl] benzoic acid ( HPF) successfully detected neutrophil-derived active oxygens ( Setsukinai K, Urano Y, Kakinuma K, Majima HJ, Nagano T. Development of novel fluorescence probes that can reliably detect reactive oxygen species and distinguish specific species. J Biol Chem 2003; 278: 3170-3175). The rate constants for the reaction of hypochlorous acid with MCI-186 and fluorescence probes was in the order of MCI-186 > APF > HPF. Fluorescence due to the oxidation of APF and HPF was observed with the stimulated neutrophils. The result that the intensity from APF oxidation was higher than that from HPF oxidation is compatible with reports that APF selectively reacts with hypochlorous acid. Fluorescence due to oxidation of both APF and HPF decreased when the reactions were carried out in the presence of a fluorescence probe and MCI-186 in a dose-dependent manner. These results indicate that MCI-186 effectively scavenges neutrophil-derived hypochlorous acid and other active oxygens.