PTS-46: THE COMET ASSAY. A METHOD TO DETERMINE THE ANTIGENOTOXIC EFFECT OF A GRAPE SEED PROCYANIDIN EXTRACT

J Pharm Pharmacogn Res 3(suppl. 1): S138, 2015

Proceedings of the 4th International Symposium on Pharmacology of Natural Products FAPRONATURA 2015  September 21st-25th, 2015; Cuban Society of Pharmacology. Topes de Collantes, Sancti Spiritus, Cuba.

Poster

PTS-46: THE COMET ASSAY. A METHOD TO DETERMINE THE ANTIGENOTOXIC EFFECT OF A GRAPE SEED PROCYANIDIN EXTRACT

Llópiz N1, Céspedes E2, Puiggros F1, Arola L1, Ardévol A1, Blade C1, Salvadó MJ1.

1Departamento de Bioquímica y Biotecnología. Universidad Rovira I Virgili, España. E-mail: niurkadariela.llopiz@urv.cat
2Facultad de Ciencias Médicas “General Calixto García”. Dirección: Ave. Universidad y J. La Habana, Cuba. E-mail: elaces@infomed.sld.cu

 

Introduction: Reactive oxygen species induce lesions in DNA and this damage is associated to a number of degenerative processes, including cancer. Various experimental methods have been proposed for measuring oxidative DNA damage. The comet assay is a very sensitive method for studying oxidative DNA damage at a single cell level. Because of that, we studied the genotoxicity of hydrogen peroxide and the effect of a grape seed procyanidin extract on DNA lesions induced by hydrogen peroxide in Fao cells. Material and Methods: Fao cells were incubated with hydrogen peroxide (50-600 μM H2O2) so that the H2O2 concentration, which induced considerable DNA damage could be found.  Cells were incubated with 600 µM H2O2 for 3 or 21 h. A procyanidin extract from grape seed (PE) was incubated or preincubated (1 h) during the exposure to H2O2. The ability of procyanidins to protect against the genotoxicity of H2O2 was compared with those of the monomeric flavanols catechin and epicatechin and the flavonol quercetin. DNA damage was monitored using the comet assay. Results: Fao cells incubated with increasing quantities of H2O2 for 3 h led to increasingly greater damage. The percentage of cells at level 3 and 4 increased to 90% at 600 µM of H2O2. PE decreased the damage caused by H2O2. The results also showed that quercetin was the most effective antioxidant of the flavonoids tested. The results indicate that procyanidins are more effective than the corresponding individual monomers, catechin and epicatechin, at preventing DNA lesions in hepatocytes and that this protection is higher after preincubation than after co-incubation. Conclusions: The protective action of the flavonoids on DNA damage may be a contribution to the antitumorigenic potential of red wine.

 

Citation Format: Llópiz N, Céspedes E, Puiggros F, Arola L, Ardévol A, Blade C, Salvadó MJ (2015) The comet assay. A method to determine the antigenotoxic effect of a grape seed procyanidin extract. [Abstract]. In: Proceedings of the FAPRONATURA 2015; 2015 Sep 21-25; Topes de Collantes, Sancti Spiritus: CSF. J Pharm Pharmacogn Res 3(Suppl. 1): S138. Abstract nr PTS-46.