Melatonin and its derivatives cyclic 3-hydroxymelatonin, N1-acetyl-N2-formyl-5-methoxykynuramine and 6-methoxymelatonin reduce oxidative DNA damage induced by Fenton reagents
Article first published online: 3 MAR 2003
Journal of Pineal Research
Volume 34, Issue 3, pages 178–184, April 2003
How to Cite
López-Burillo, S., Tan, D.-X., Rodriguez-Gallego, V., Manchester, L. C., Mayo, J. C., Sainz, R. M. and Reiter, R. J. (2003), Melatonin and its derivatives cyclic 3-hydroxymelatonin, N1-acetyl-N2-formyl-5-methoxykynuramine and 6-methoxymelatonin reduce oxidative DNA damage induced by Fenton reagents. Journal of Pineal Research, 34: 178–184. doi: 10.1034/j.1600-079X.2003.00025.x
- Issue published online: 3 MAR 2003
- Article first published online: 3 MAR 2003
- Received September 5, 2002; accepted November 4, 2002.
- N 1-acetyl-N 2-formyl-5-methoxykynuramine;
- cyclic 3-hydroxymelatonin;
- 8-hydroxydeoxy- guanosine;
- hydroxyl radical;
Abstract: Free radicals are generated in vivo and they oxidatively damage DNA because of their high reactivities. In the last several years, hundreds of publications have confirmed that melatonin is a potent endogenous free radical scavenger. Some of the metabolites produced as a result of these scavenging actions have been identified using pure chemical systems. This is the case with both N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK), identified as a product of the scavenging reaction of H2O2 by melatonin, and cyclic 3-hydroxymelatonin (C-3-OHM) which results when melatonin detoxifies two hydroxyl radicals (ḃOH). In the present in vitro study, we investigated the potential of two different derivatives of melatonin to scavenger free radicals. One of these derivatives is C-3-OHM, while the other is 6-methoxymelatonin (6-MthM). We also examined the effect of two solvents, i.e., methanol and acetonitrile, in this model system. As an endpoint, using high-performance liquid chromatography we measured the formation of 8-hydroxy-2′-deoxyguanosine (8-OH-dG) in purified calf thymus DNA treated with the Fenton reagents, chromium(III) [Cr(III)] plus H2O2, in the presence and in the absence of these molecules. The 8-OH-dG is considered a biomarker of oxidative DNA damage. Increasing concentrations of Cr(III) (as CrCl3) and H2O2 was earlier found to induce progressively greater levels of 8-OH-dG in isolated calf thymus DNA because of the generation of ḃOH via the Fenton-type reaction. We found that C-3-OHM reduces ḃOH-mediated damage in a dose-dependent manner, with an IC50 = 5.0 ± 0.2 μm; melatonin has an IC50 = 3.6 ± 0.1 μm. These values differ statistically significantly with P < 0.05. In these studies, AFMK had an IC50 = 17.8 ± 0.7 μm (P < 0.01). The 6-MthM also reduced DNA damage in a dose-dependent manner, with an IC50 = 4.2 ± 0.2 μm; this value does not differ from the ICs for melatonin and C-3-OHM. We propose a hypothetical reaction pathway in which a mole of C-3-OHM scavenges 2 mol of ḃOH yielding AFMK as a final product. As AFMK is also a free radical scavenger, the action of melatonin as a free radical scavenger is a sequence of scavenging reactions in which the products are themselves scavengers, resulting in a cascade of protective reactions.