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Butylated hydroxyanisole (BHA) is a synthetic phenolic compound consisting of a mixture of two isomeric organic compounds: 2-tert-butyl-4-hydroxyanisole and 3-tert-butyl-4-hydroxyanisole. We examined the effect of BHA against hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-induced apoptosis in primary cultured mouse hepatocytes. Cell viability was significantly decreased by H<sub>2</sub>O<sub>2</sub> in a dose-dependent manner. Additionally, H<sub>2</sub>O<sub>2</sub> treatment increased Bax, decreased Bcl-2, and promoted PARP-1 cleavage in a dose-dependent manner. Pretreatment with BHA before exposure to H<sub>2</sub>O<sub>2</sub> significantly attenuated the H<sub>2</sub>O<sub>2</sub>-induced decrease of cell viability. H<sub>2</sub>O<sub>2</sub> exposure resulted in an increase of intracellular reactive oxygen species (ROS) generation that was significantly inhibited by pretreatment with BHA or N-acetyl-cysteine (NAC, an ROS scavenger). H<sub>2</sub>O<sub>2</sub>-induced decrease of cell viability was also attenuated by pretreatment with BHA and NAC. Furthermore, H<sub>2</sub>O<sub>2</sub>-induced increase of Bax, decrease of Bcl-2, and PARP-1 cleavage was also inhibited by BHA. Taken together, results of this investigation demonstrated that BHA protects primary cultured mouse hepatocytes against H<sub>2</sub>O<sub>2</sub>-induced apoptosis by inhibiting ROS generation.

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