Ji Jian, Zhu Pei, Sun Chao, Sun Jiadi, An Lu, Zhang Yinzhi, Sun Xiulan
State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, China.
J Toxicol Sci. 2017;42(1):43-52. doi: 10.2131/jts.42.43.
3-Chloropropane-1,2-diol (3-MCPD) is a heat-produced contaminant formed during the preparation of soy sauce worldwide. The present investigation was conducted to determine the molecular aspects of 3-MCPD toxicity on human embryonic kidney cells (HEK293). Cell viability and apoptosis were assessed in response to exposure to 3-MCPD using the MTT assay and high-content screening (HCS). DNA damage, intracellular reactive oxygen species (ROS) and apoptosis-related proteins were evaluated. Genes related with apoptosis were detected by qPCR-array for further understanding the 3-MCPD induced cell apoptosis signaling pathway. Our results clearly showed that 3-MCPD treatment inhibits cell proliferation and reactive oxygen species generation. qPCR-array indicated that nine apoptotic genes were up-regulated more than 2-fold and six down-regulated more than 2-fold. Genes associated with the mitochondrial apoptotic pathway, especially BCL2 family genes, changed significantly, indicating that the mitochondrial apoptotic pathway is activated. Death receptor pathway-related genes, TNFRSF11B and TNFRSF1A, changed significantly, indicating that the death receptor pathway is also activated, resulting in the inhibition of cell growth and proliferation as well as induction of apoptosis. To sum up, the experiment results indicated that 3-MCPD induced HEK293 cell toxicity through the death receptor pathway and mitochondrial pathway.
3-氯-1,2-丙二醇（3-MCPD）是一种在全球酱油制备过程中产生的热致污染物。本研究旨在确定3-MCPD对人胚肾细胞（HEK293）毒性的分子机制。使用MTT法和高内涵筛选（HCS）评估3-MCPD暴露后细胞活力和凋亡情况。评估DNA损伤、细胞内活性氧（ROS）和凋亡相关蛋白。通过qPCR阵列检测与凋亡相关的基因，以进一步了解3-MCPD诱导的细胞凋亡信号通路。我们的结果清楚地表明，3-MCPD处理可抑制细胞增殖和活性氧生成。qPCR阵列表明，九个凋亡基因上调超过2倍，六个下调超过2倍。与线粒体凋亡途径相关的基因，尤其是BCL2家族基因，发生了显著变化，表明线粒体凋亡途径被激活。死亡受体途径相关基因TNFRSF11B和TNFRSF1A发生了显著变化，表明死亡受体途径也被激活，导致细胞生长和增殖受到抑制以及凋亡诱导。综上所述，实验结果表明了3-MCPD通过死亡受体途径和线粒体途径诱导HEK293细胞毒性。
