The investigation of the possible effects of resveratrol on human coronary artery endothelial cell damage induced by in vitro hydrogen peroxide

Introduction: Oxidative stress is defined as an imbalance between the production and destruction of reactive oxygen species. Hydrogen peroxide resulting from vascular and inflammatory cells causes endothelial dysfunction by inducing oxidative stress, which is of importance in coronary artery disease, cardiac failure, hypertension, peripheral artery disease and diabetes. Defense sources of enzymatic antioxidant include superoxide dismutase, glutathione peroxidase and glutathione catalase. Non-enzymatic antioxidants are ascorbic acid, ?-tocopherol, glutathione, carotenoids, flavonoids, stilbenes and others. Resveratrol, a subclass of stilbenes family, is a polyphenolic compound found in grapes and red wine. It has anti-inflammatory, anti-oxidant, anti-apoptotic, cytoprotective, anti-cancer and cardio-protective effects.Aim: No research conducted on in vitro effects of resveratrol on endothelial cells induced by hydrogen peroxide damage have been found in the literature. The purpose of this study is to investigate whether resveratrol can protect in vitro human endothelial cells against hydrogen peroxide damage. Therefore, cell cytotoxicity, production of reactive oxygen species, expressions of ?-glutamylcysteine synthetase, glutathione reductase and glutathione peroxidase enzymes and glutathione levels were investigated.Materials and methods: In this study, various concentrations of resveratrol found in in vitro human coronary endothelial cells and the effects of resveratrol on cell cytotoxicity induced by hydrogen peroxide during pre-incubation periods were investigated. The effect of resveratrol on cell damage was investigated with the lactate dehydrogenase test, a test used for the evaluation of cell cytotoxicity. The effects of various concentrations of resveratrol on the production of reactive oxygen species during various pre-incubation periods and time slices were evaluated by using hydroxyphenyl fluorescein, an oxidant-sensitive fluorescent probe. Anti-oxidant effects of resveratrol, protein expressions of ?-glutamylcysteine synthetase, glutathione peroxidase and glutathione reductase enzymes were evaluated with the Western blot method. The effects of resveratrol on the glutathione level in vitro human coronary endothelial cells were evaluated with high performance liquid chromatography.Results: It was determined that 10 and 50 ? M concentrations of resveratrol statistically significantly decreased the cytotoxicity induced by 750 ? M hydrogen peroxide during 1, 12 and 24-hour pre-icubation periods for one hour (p<0.05). It was also determined that 10, 50 and 100 ? M concentrations of resveratrol statistically significantly decreased the production of reactive oxygen species at 15th and 60th minutes and 6th hour during the 24-hour pre-icubation periods (p<0.05). Resveratrol statistically significantly increases the reduced glutathione levels (p<0.05). Resveratrol increases the protein expressions of ?-glutamylcysteine synthetase, glutathione peroxidase-1 and glutathione reductase enzymes (p<0.05).Conclusion: All the data support each other and suggest that resveratrol has a protective effect on the human coronary artery endothelial cell damage. It is thought that the findings obtained in this study can light the way to the new therapeutic approaches to be used for the reversal of oxidative stress which develops in cardiovascular diseases.Key words: Resveratrol, human coronary artery endothelial cell, oxidative damage