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8493717 
Journal Article 
Butylated hydroxytoluene, a food additive, modulates membrane potential and increases the susceptibility of rat thymocytes to oxidative stress 
Kamemura, N 
2018 
1 
Computational Toxicology
ISSN: 2468-1113 
Elsevier B.V. 
6 
32-38 
English 
Butylated hydroxytoluene (BHT) is a food additive with an antioxidant activity. BHT is a weak antioxidant in vegetable oils and is often added to meat fats. In addition, BHT is commonly used as a dog food additive. Toxicity of BHT has been reported under in vitro and in vivo experimental conditions. However, the mechanism of BHT-induced toxic effect in cells is unclear. In the present study, we examined the toxic effect of BHT and the cellular parameters affected by BHT in rat thymocytes by performing flow cytometry analysis with fluorescent probes. BHT at 10 significantly increased the population of cells exhibiting propidium fluorescence, i.e. the population of dead cells in time-dependent manner. Treatment of rat thymocytes with 3–10 µM BHT depolarized the cell membrane and decreased cellular non-protein thiol content. Treatment with 10 µM BHT increased intracellular Ca2+ and Zn2+ concentrations and cell lethality. Moreover, treatment with 10 µM BHT increased the population of dead cells irrespective of phosphatidylserine expression on the outer membrane. Cotreatment with 10 µM BHT and 100 µM H2O2 synergistically increased cell lethality. Our results indicate that the mechanism of BHT-induced toxicity was to be increased intracellular Ca2+ and Zn2+ concentrations, resulting in increased susceptibility of rat thymocytes to oxidative stress. © 2018 
Butylated hydroxytoluene; Cytotoxicity; Food additive; Oxidative stress