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.

Graphical Abstract

中文翻译：

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双酚 A 诱导太平洋红鲷鱼白细胞中活性氧的产生和细胞凋亡相关基因的表达

双酚A是世界上聚碳酸酯塑料工业中最常用的成分之一。这种污染物对鱼类的体外和体内细胞具有破坏作用。本研究首次评估了双酚 A (BPA) 对从太平洋红鲷鱼Lutjanus peru分离的头肾和脾白细胞诱导的细胞毒性、氧化应激和细胞凋亡。头肾和脾脏白细胞在 2 小时和 24 小时分别暴露于 100、1000 和 10,000 µg/mL 的 BPA。结果显示 BPA 在 1000 和 10,000 µg/mL 时具有细胞毒性。在暴露于 100 µg/mL 2 小时的白细胞中观察到细胞活力 > 80%；因此，该浓度被选择用于剩余的研究。通过 DCF-DA 和 NBT 测定分析，2 或 24 小时后，与对照组相比，暴露于 BPA 的白细胞中活性氧 (ROS) 的产生显着增加。接触 BPA 24 小时后，头肾白细胞中的超氧化物歧化酶和过氧化氢酶活性增加。根据头肾白细胞中半胱天冬酶（casp-1和casp-3）、颗粒酶 A ( granz-A ) 和穿孔素 1 ( perf-1 ) 基因表达推断细胞凋亡，在 BPA 暴露 2 小时后，这些基因表达显着上调，以及来自granz-A和perf-1 的基因，在脾脏白细胞中暴露于 BPA 24 小时后，它们的表达上调。通过扫描电子显微镜观察到短的细胞质延长和膜泡，提示细胞凋亡。这些数据表明，100 µg/mL 的 BPA 会诱导太平洋红鲷鱼头肾和脾白细胞的细胞毒性、氧化应激和细胞凋亡。

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