Comparison of superoxide detection abilities of newly developed spin traps in the living cells
Comparison of superoxide detection abilities of newly developed spin traps in the living cells
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DOI：
10.1080/10715760902988850
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发表时间：
2009-01-01
影响因子：
3.3
通讯作者：
Kohno, Masahiro
中科院分区：
文献类型：
--
作者：
Saito, Keita;Takahashi, Miho;Kohno, Masahiro
本研究比较了5，5-二甲基-1-吡咯啉-N-氧化物(DMPO)、5-(二乙氧基磷酰基)-5-甲基-1-吡咯啉-N-氧化物(DEPMPO)、5-(二苯基膦)-5-甲基-1-吡咯啉-氧化物(DPPMPO)和5-(2，2-二甲基-1，3-丙氧基-环磷酰基)-5-甲基-1-吡咯啉N-氧化物(CYPMPO)四种自旋陷阱对活细胞中超氧阴离子的检测能力。在佛波酯刺激的人口腔多形核白细胞悬液中加入自旋陷阱，观察到超氧化物加合物的电子自旋共振(ESR)信号。CPPMPO-超氧化物加合物(CYPMPO-OHO)的ESR信号在反应开始后24分钟增强，而DMPO-OHO和DPPMPO-OOH的ESR信号分别在6分钟和10分钟达到峰值。OPMN中DMPO-OHO、DPPMPO-OHO和CYPMPO-OOH的最大浓度分别为1.9、6.0和10.7 mU M。此外，与DEPMPO相比，CYPMPO能更有效地捕获血液中PMN中的超氧化物。从这些结果可以得出结论，在活细胞系统中，由于细胞毒性较低，其超氧化物加合物寿命较长，因此，在测量活细胞系统中的超氧化物时，CYPMPO比DMPO、DPPMPO和DEPMPO表现得更好。
This study compared the superoxide detection abilities of four spin traps, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide (DEPMPO), 5-(diphenylphosphinoyl)-5-methyl-1pyrroline N-oxide (DPPMPO) and 5-(2,2-dimethyl-1,3-propoxy cyclophosphoryl)-5-methyl-1-pyrroline N-oxide (CYPMPO) in living cells. Electron spin resonance (ESR) signals of the superoxide adducts were observed when spin traps were added to a suspension of human oral polymorphonuclear leukocytes (OPMNs) stimulated by phorbol 12-myristate 13-acetate. The ESR signal of the CYPMPO-superoxide adduct (CYPMPO-OOH) increased for 24 min after the initiation of the reaction, whereas the signals from DMPO-OOH and DPPMPO-OOH peaked at 6 and 10 min, respectively. The maximum concentrations of DMPO-OOH, DPPMPO-OOH and CYPMPO-OOH in OPMNs were 1.9, 6.0 and 10.7 mu M, respectively. Furthermore, CYPMPO could more efficiently trap superoxide in blood PMNs compared with DEPMPO. From these results, it was concluded that CYPMPO performs better than DMPO, DPPMPO and DEPMPO for superoxide measurements in living cell systems because it has lower cytotoxicity and its superoxide adduct has a longer lifetime.