Electron spin resonance spectroscopic demonstration of the hydroxyl free radical scavenger properties of dimethylaminoethanol in spin trapping experiments confirming the molecular basis for the biological effects of centrophenoxine

Abstract

The ADP-Fe(II)-H 2O 2 system generates OH free radicals which can be trapped by 5,5-dimethyl-1-pyrroline- N-oxide (DMPO) thus yielding a measurable signal by electron spin resonance spectroscopy. The amount of DMPO-OH spin adduct formed under certain conditions decreased considerably, if dimethylaminoethanol (DMAE), p-chlorophenoxyacetic acid (PCPA) or centrophenoxine (CPH) were present in comparable concentrations to that of DMPO. It has been demonstrated that such an effect cannot be attributed to any interference of the tested compounds with the Fe(II) and its oxidability by H 2O 2. The reaction of DMAE with OH free radicals was demonstrated also by using other spin traps. These spin traps reacted with OH free radicals either not at all (phenyl-tert-butylnitrone, PBN) or only to a slight extent (alfa-pyridyl-1-oxide- N-tert-butylnitrone, 4-POBN). DMAE was also a competitive OH free radical scavenger with proline and hydroxyproline, both of which have recently been shown to react with OH free radicals to form nitroxyl free radicals. On the basis of the experimental results, the OH free radical scavenger property of DMAE can be regarded as firmly established. This result supports the molecular mechanism proposed for the explanation of the anti-aging effects of CPH in terms of the membrane hypothesis of aging.