Protective effect of esculetin against oxidative stress-induced cell damage via scavenging reactive oxygen species

Aim: To investigate the anti-oxidant properties of esculetin (6,7-dihydro-xycoumarin) against H(2)O(2)-induced Chinese hamster lung fibroblast (V79-4) damage. Methods: The radical scavenging activity was assessed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, hydroxyl radical, and intracellular reactive oxygen species (ROS). In addition, lipid peroxidation was assayed by the measure of related substances which react with thiobarbituric acid. The amount of carbonyl formation in protein was determined using a protein carbonyl ELISA kit. As well, cellular DNA damage was detected by Western blot and immuno-fluorescence image. Cell viability was assessed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: Esculetin exhibited DPPH radical scavenging, hydroxyl radical scavenging, and intracellular ROS scavenging activities. The radical scavenging activity of esculetin resulted in the protection of cells from lipid peroxidation, protein carbonyl, and DNA damage induced by H(2)O(2). Therefore, esculetin recovered cell viability exposed to H(2)O(2). Conclusion: Esculetin efficiently attenuated the oxidative stress induced cell damage via its anti-oxidant properties. As a result, esculetin may be useful in the development of functional food and raw materials of medicine.