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Original

Comparison of superoxide detection abilities of newly developed spin traps in the living cells

, , , &
Pages 668-676 | Received 18 Dec 2008, Published online: 21 Jul 2009
 

Abstract

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 µM, 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.

Abbreviations
BPMN=

blood polymorphonuclear leukocyte

CYPMPO=

5-(2,2-dimethyl-1,3-propoxy cyclophosphoryl)-5-methyl-1-pyrroline N-oxide

DEPMPO=

5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide

DMPO=

5,5-dimethyl-1-pyrroline-N-oxide

DMSO=

dimethyl sulphoxide

DPPMPO=

5-(diphenylphosphinoyl)-5-methyl-1-pyrrole N-oxide

DTPA=

diethylenetriamine-N,N,N′,N″,N″-pentaacetic acid

ESR=

electron spin resonance

HPX=

hypoxanthine

OPMN=

oral polymorphonuclear leukocyte

ROS=

reactive oxygen species

SOD=

superoxide dismutase

XOD=

xanthine oxidase

Abbreviations
BPMN=

blood polymorphonuclear leukocyte

CYPMPO=

5-(2,2-dimethyl-1,3-propoxy cyclophosphoryl)-5-methyl-1-pyrroline N-oxide

DEPMPO=

5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide

DMPO=

5,5-dimethyl-1-pyrroline-N-oxide

DMSO=

dimethyl sulphoxide

DPPMPO=

5-(diphenylphosphinoyl)-5-methyl-1-pyrrole N-oxide

DTPA=

diethylenetriamine-N,N,N′,N″,N″-pentaacetic acid

ESR=

electron spin resonance

HPX=

hypoxanthine

OPMN=

oral polymorphonuclear leukocyte

ROS=

reactive oxygen species

SOD=

superoxide dismutase

XOD=

xanthine oxidase

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