The present work describes the protective effects of thymol isolated from Thymus quinquecostatus Celak. against tert-butyl hydroperoxide (t-BHP)-induced oxidative damage through various experiments with Chang liver cells. Thymol significantly protected hepatocytes against t-BHP-induced cell cytotoxicity as demonstrated by increased viability. Furthermore, observation of Hoechst staining, annexin V/PI staining, and expression of Bcl-2 and Bax indicated that thymol inhibited t-BHP-induced Chang cell damage. Further, thymol inhibited the loss of mitochondrial membrane potential in t-BHP-treated Chang cells and prevented oxidative stress-triggered reactive oxygen species (ROS) and lipid peroxidation (malondialdehyde, MDA). Thymol restored the antioxidant capability of hepatocytes including glutathione (GSH) levels which were reduced by t-BHP. These results indicated that thymol prevents oxidative stress-induced damage to liver cells through suppression of ROS and MDA levels and increase of GSH level.