Antioxidative effect of melatonin on DNA and erythrocytes against free-radical-induced oxidation

The aim of this work is to investigate the antioxidative effect of melatonin (N-acetyl-5-methoxytryptamine) on the oxidation of DNA and human erythrocytes induced by 2,2′-azobis(2-amidinopropane hydrochloride) (AAPH). First, the 50% inhibition concentration (IC50) of melatonin is measured by reacting with two radical species, i.e., 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS+) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH). The IC50 of melatonin are 75 μM and 300 μM when melatonin reacts with ABTS+ and DPPH, respectively. Especially, the reactions of melatonin with ABTS+ and DPPH are the direct evidence for melatonin to trap radicals. Then, melatonin is applied to protect DNA and human erythrocytes against oxidative damage and hemolysis induced by 2,2′-azobis(2-amidinopropane hydrochloride) (AAPH). The presence of melatonin prolongs the occurrence of the oxidative damage of DNA and hemolysis of erythrocytes, generating an inhibition period (tinh). The proportional relationship between tinh and the concentration of melatonin ([MLT]) is treated by the chemical kinetic equation, tinh = (n/Ri)[MLT], in which n means the number of peroxyl radical trapped by an antioxidant, and Ri stands for the initiation rate of the radical reaction. It is found that every molecule of melatonin can trap almost two radicals in protecting DNA and erythrocytes. Furthermore, quantum calculation proves that the indole-type radical derived from melatonin is much stable than amide-type radical. Finally, melatonin is able to accelerate hemolysis of erythrocytes induced by hemin, indicating that melatonin leads to the collapse of the erythrocyte membrane in the presence of hemin. This may provide detailed information for the usage of melatonin and helpful reference for the design of indole-related drugs.