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Ochratoxin A-Induced Apoptosis of IPEC-J2 Cells through ROS-Mediated Mitochondrial Permeability Transition Pore Opening Pathway

Wang, Hong, Chen, Ying, Zhai, Nianhui, Chen, Xingxiang, Gan, Fang, Li, Hu, Huang, Kehe
Journal of agricultural and food chemistry 2017 v.65 no.48 pp. 10630-10637
Western blotting, apoptosis, caspase-3, cell lines, confocal microscopy, cyclosporine, cytochrome c, cytotoxicity, epithelial cells, flow cytometry, intestinal mucosa, mitochondria, models, ochratoxin A, permeability, reactive oxygen species, staining, swine
With the purpose to explore the mechanisms associated with the intestinal toxicity of Ochratoxin A (OTA), an intestinal porcine epithelial cell line (IPEC-J2) was applied in this study as in vitro models for intestinal epithelium. The results confirmed that OTA induced IPEC-J2 cell toxicity by MTT assay and apoptosis by Hoechst 33258 staining and flow cytometer analysis. We also observed that OTA induced the mitochondrial reactive oxygen species (ROS) production and mitochondrial permeability transition pore (mPTP) opening by confocal microscopy. Western blot showed that OTA induced cytochrome c (cyt-c) release and caspase-3 activation, which could be suppressed by inhibition of mPTP opening with cyclosporin A. Treatment with Mito-TEMPO, the mitochondria-targeted ROS scavenger, blocked OTA-induced mitochondrial ROS generation and mPTP opening and prevented cyt-c release, caspase-3 activation, and apoptosis in IPEC-J2 cells.