PMID- 27363784 OWN - NLM STAT- MEDLINE DCOM- 20170207 LR - 20170926 IS - 1879-3169 (Electronic) IS - 0378-4274 (Linking) VI - 258 DP - 2016 Sep 6 TI - T-2 toxin inhibits murine ES cells cardiac differentiation and mitochondrial biogenesis by ROS and p-38 MAPK-mediated pathway. PG - 259-266 LID - S0378-4274(16)32252-4 [pii] LID - 10.1016/j.toxlet.2016.06.2103 [doi] AB - OBJECTIVE: To investigate the effect of T-2 toxin on murine embryonic stem cells (ESCs) cardiac differentiation and mitochondrial biogenesis in vitro. METHODS: Cardiac differentiation of the mouse ESCs was initiated by embryoid bodies (EBs) formation in hanging drops. EBs were exposed to 0.5ng/ml T-2 toxin for 24, 72 and 120h. Cultures were observed daily for the appearance of contracting clusters, and cardiac-specific protein (alpha-actiniin) were measured by Western blot and immunocytochemistry. Mitochondrial ultrastructure was observed by confocal laser scanning microscopy and transmission EM photography. Reactive oxygen species (ROS) was monitored by H2-dichlorofluorescein-diacetate (H2DCF-DA). The phosphorylation of the p38 (p-p38) and p38 mitogen-activated protein kinase (MAPK) and the expression of mitochondrial biogenesis proteins, including peroxisome proliferator activated receptor coactivator-1 alpha (PGC-1alpha), nuclear respiratory factor 1 (NRF-1), mitochondrial transcription factor A (mtTFA), and mitochondrial respiratory chain complex IV (COXIV) were analyzed using Western blot. In some experiments, mESCs were pre-treated with the antioxidant Trolox (200muM) for 30min, then exposed to Trolox (200muM) and T-2 toxin (0.5ng/ml) for 72h. RESULTS: Contracting clusters were observed under the microscope light and cardiac-specific protein (alpha-actinin) expressed positively indicated mESCs directly differentiated in cardiomyocytes. However, the cardiac differentiation was inhibited by T-2 toxin treatment 72 and 120h. ROS accumulated in murine ES cells in a time-dependent manner. The expression of p-p38 significantly increased in 24h group and decrease in 72 and 120h groups. The decrease of mitochondrial number and the mitochondrial biogenesis-related proteins expression, including PGC-1alpha, NRF-1, mtTFA, and COXIV decreased in a time-dependent manner with T-2 toxin treatment. However, the inhibition of mitochondrial biogenesis by T-2 toxin in differentiated mESCs was recovered significantly in the presence of the antioxidant Trolox. CONCLUSION: Taken together, T-2 toxin decreased the expression of PGC-1alpha, NRF-1, and mtTFA, inhibited mitochondrial biogenesis, and then inhibited the cardiac differentiation of murine ES cells, and the effect was partly responsible for the p38 MAPK mediated by ROS. CI - Copyright (c) 2016 Elsevier Ireland Ltd. All rights reserved. FAU - Fang, Haiqin AU - Fang H AD - Evaluation and Research Centre for Toxicology, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China; Key Laboratory of Food Safety Risk Assessment of Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China. FAU - Cong, Liangzi AU - Cong L AD - Huaiyin District Center for Disease Control and Prevention, Jinan, Shandong, China. FAU - Zhi, Yuan AU - Zhi Y AD - Key Laboratory of Food Safety Risk Assessment of Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China. FAU - Xu, Haibin AU - Xu H AD - Key Laboratory of Food Safety Risk Assessment of Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China. FAU - Jia, Xudong AU - Jia X AD - Key Laboratory of Food Safety Risk Assessment of Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China. Electronic address: jiaxudong@cfsa.net.cn. FAU - Peng, Shuangqing AU - Peng S AD - Evaluation and Research Centre for Toxicology, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, China. Electronic address: pengsq@hotmail.com. LA - eng PT - Journal Article DEP - 20160627 PL - Netherlands TA - Toxicol Lett JT - Toxicology letters JID - 7709027 RN - 0 (Antioxidants) RN - 0 (Biomarkers) RN - 0 (Reactive Oxygen Species) RN - 0 (Teratogens) RN - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases) RN - I3FL5NM3MO (T-2 Toxin) SB - IM MH - Animals MH - Antioxidants/pharmacology MH - Biomarkers/metabolism MH - Cell Differentiation/drug effects MH - Cell Line MH - Coculture Techniques MH - MAP Kinase Signaling System/drug effects MH - Mice MH - Mitochondrial Dynamics/*drug effects MH - Mouse Embryonic Stem Cells/cytology/*drug effects/metabolism/ultrastructure MH - Myoblasts, Cardiac/cytology/*drug effects/metabolism/ultrastructure MH - Myocytes, Cardiac/cytology/*drug effects/metabolism/ultrastructure MH - Organelle Biogenesis MH - Oxidative Stress/*drug effects MH - Phosphorylation/drug effects MH - Protein Processing, Post-Translational/drug effects MH - Reactive Oxygen Species/agonists/antagonists & inhibitors/metabolism MH - T-2 Toxin/antagonists & inhibitors/*toxicity MH - Teratogens/chemistry/*toxicity MH - p38 Mitogen-Activated Protein Kinases/metabolism OTO - NOTNLM OT - Cardiac differentiation OT - Mitochondrial biogenesis OT - PGC-1alpha OT - ROS OT - T-2 toxin OT - p38MAPK EDAT- 2016/07/02 06:00 MHDA- 2017/02/09 06:00 CRDT- 2016/07/02 06:00 PHST- 2016/04/19 00:00 [received] PHST- 2016/06/19 00:00 [revised] PHST- 2016/06/26 00:00 [accepted] PHST- 2016/07/02 06:00 [entrez] PHST- 2016/07/02 06:00 [pubmed] PHST- 2017/02/09 06:00 [medline] AID - S0378-4274(16)32252-4 [pii] AID - 10.1016/j.toxlet.2016.06.2103 [doi] PST - ppublish SO - Toxicol Lett. 2016 Sep 6;258:259-266. doi: 10.1016/j.toxlet.2016.06.2103. Epub 2016 Jun 27.