Bisphenol A Induces Reactive Oxygen Species Production and Apoptosis-Related Gene Expression in Pacific Red Snapper Lutjanus peru Leukocytes

Bisphenol A is one of the most used components of the polycarbonate plastic industry in the word. This contaminant has disrupting effect in cells in in vitro and in vivo in fish. This study evaluated for the first time the cytotoxicity, oxidative stress and apoptosis induced by bisphenol A (BPA) in head-kidney and spleen leukocytes isolated from Pacific red snapper Lutjanus peru. Head-kidney and spleen leukocytes were exposed to 100, 1000 and 10,000 µg/mL of BPA at 2 and 24 h. Results showed cytotoxicity of BPA at 1000 and 10,000 µg/mL. Cell viability > 80% was observed in leukocytes exposed to 100 µg/mL for 2 h; thus, this concentration was selected for the remainder of the study. Reactive oxygen species (ROS) production, analyzed by DCF-DA and NBT assays, significantly increased in those leukocytes exposed to BPA compared to controls after 2 or 24 h. Superoxide dismutase and catalase activities increased in head-kidney leukocytes after 24 h of BPA exposure. Apoptosis was inferred from caspase (casp-1 and casp-3), granzyme A (granz-A) and perforin 1 (perf-1) gene expression, which was significantly up-regulated, at 2 h BPA exposure in head-kidney leukocytes, and from granz-A and perf-1, which were up-regulated, after 24 h BPA exposure in spleen leukocytes. Short cytoplasmic prolongations and membrane blebs, suggestive of apoptosis, were observed by scanning electron microscopy. These data suggest that BPA at 100 µg/mL induces cytotoxicity, oxidative stress, apoptosis in Pacific red snapper head-kidney and spleen leukocytes.