T-2 toxin induces apoptosis in differentiated murine embryonic stem cells through reactive oxygen species-mediated mitochondrial pathway
T-2 toxin induces apoptosis in differentiated murine embryonic stem cells through reactive oxygen species-mediated mitochondrial pathway
复制标题
DOI：
10.1007/s10495-012-0724-3
复制
发表时间：
2012-08-01
期刊：
影响因子：
7.2
通讯作者：
Peng, Shuangqing
中科院分区：
文献类型：
--
作者：
Fang, Haiqin;Wu, Yingliang;Peng, Shuangqing
T-2毒素是镰刀菌产生的毛霉烯类真菌毒素家族的一员，已被证明在体内对多个靶点具有多种毒性作用。然而，T-2毒素的体外胚胎毒性尚不清楚。本研究使用胚胎干细胞(ES细胞D3)和成纤维细胞3T3细胞两种永久性细胞系来评价T-2毒素的毒性。将分化的小鼠ES细胞与不同浓度的T-2毒素(0.5、1和2 ng/ml)共同培养24 h，观察到1和2 ng/ml的T-2毒素诱导的细胞内ROS、脂质和DNA氧化损伤以及线粒体跨膜电位的丧失。流式细胞仪检测显示T-2毒素诱导细胞周期停滞和细胞凋亡。此外，T-2毒素开放线粒体通透性转换孔，引起线粒体细胞色素c释放，诱导P53、caspase-9、caspase-3表达上调，Bax/Bcl2比值升高。然而，在抗氧化剂Trolox的存在下，T-2毒素诱导的分化的ES细胞的氧化损伤和凋亡显著减少。综上所述，这些结果表明T-2毒素诱导分化的小鼠ES细胞氧化应激和凋亡，而ROS介导的线粒体途径在T-2毒素诱导的细胞凋亡中起重要作用。
T-2 toxin, a member of the trichothecene mycotoxin family produced by the Fusarium fungi, has been shown to exert a variety of toxic effects on multiple targets in vivo. However, the embryonic toxicity of T-2 toxin in vitro remains unclear. In the present study, two permanent cell lines, embryonic stem cells (ES cells D3) and fibroblast 3T3 cells, were used to evaluate T-2 toxin toxicity. Differentiated mouse ES cells were cultivated as embryoid bodies along with T-2 toxin at different concentrations (0.5, 1, and 2 ng/ml) for 24 h. The increases in cellular reactive oxygen species (ROS), lipid and DNA oxidative damage, and loss of mitochondrial transmembrane potential were observed at 1 and 2 ng/ml concentrations. Flow cytometry showed that T-2 toxin induced cell cycle arrest and apoptosis. Furthermore, T-2 toxin opened the mitochondrial permeability transition pore, caused the release of cytochrome c from mitochondria and induced the upregulation of p53, caspase-9, caspase-3 expression and increased the ratio of Bax/Bcl-2. However, T-2 toxin-induced oxidative damage and apoptosis in differentiated ES cells decreased significantly in the presence of the antioxidant Trolox. Taken together, these results demonstrate that T-2 toxin induces oxidative stress and apoptosis in differentiated murine ES cells, and ROS-mediated mitochondrial pathway plays an important role in T-2 toxin induced apoptosis.