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Impact of 2-bromopropane on mouse embryonic stem cells and related regulatory mechanisms

机译：2-溴丙烷对小鼠胚胎干细胞的影响及相关调控机制

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2-Bromopropane (2-BP), a cleaning agent, is used as an alternative to ozone-depleting solvents. Previously, 2-BP was shown to have cytotoxic effects on mouse blastocysts and is associated with defects in their subsequent development, both?in vitro?and?in vivo. In addition, it was found that 2-BP also has cytotoxic effects on oocyte maturation and subsequent pre- and post implantation development?in vitroand?in vivo, and significantly reduces the rate of oocyte maturation, fertilization, and embryonic development?in vitro. This study shows that 2-BP (5 to 10 μM) induces apoptotic processes in mouse embryonic stem cells (ESC-B5), but exerts no effects at treatment dosages below 5 μM. In ESC-B5 cells, 2-BP directly increased the content of reactive oxygen species (ROS), significantly increased the cytoplasmic free calcium and nitric oxide (NO) levels, triggered a loss of mitochondrial membrane potential (MMP), activated caspases-9 and -3, and induced cell death. Pre-treatment with NO scavengers suppressed the apoptotic biochemical changes induced by 10 μM 2-BP and promoted the gene expression levels of p53 and p21, which are involved in apoptotic signaling. These results demonstrate for the first time that 2-BP triggers apoptosis in mouse embryonic stem cells via ROS, NO and the activation of mitochondria-dependent cell death signaling.
机译：2-溴丙烷（2-BP）是一种清洁剂，可替代消耗臭氧的溶剂。以前，2-BP被证明对小鼠胚泡有细胞毒性作用，并且与它们随后的体外和体内发育缺陷有关。另外，还发现2-BP对卵母细胞成熟以及随后的体内和体外植入前后的发育也具有细胞毒性作用，并显着降低了体外卵母细胞的成熟，受精和胚胎发育的速率。这项研究表明，2-BP（5至10μM）诱导小鼠胚胎干细胞（ESC-B5）的凋亡过程，但在低于5μM的治疗剂量下无作用。在ESC-B5细胞中，2-BP直接增加了活性氧（ROS）的含量，显着增加了细胞质中游离钙和一氧化氮（NO）的水平，触发了线粒体膜电位（MMP）的丧失，激活了caspases-9 -3和诱导细胞死亡。 NO清除剂的预处理抑制了10μM2-BP诱导的凋亡生化变化，并促进了参与凋亡信号转导的p53和p21基因表达水平。这些结果首次证明2-BP通过ROS，NO和线粒体依赖性细胞死亡信号的激活触发小鼠胚胎干细胞的凋亡。

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