OTA induces intestinal epithelial barrier dysfunction and tight junction disruption in IPEC-J2 cells through ROS/Ca2+-mediated MLCK activation

被引:71
作者
Wang, Hong [1 ,2 ]
Zhai, Nianhui [1 ,2 ]
Chen, Ying [1 ,2 ]
Fu, Chongyang [1 ,2 ]
Huang, Kehe [1 ,2 ]
机构
[1] Nanjing Agr Univ, Coll Vet Med, Nanjing 210095, Jiangsu, Peoples R China
[2] Nanjing Agr Univ, Inst Nutr & Metab Disorders Domest Anim & Fowls, Nanjing 210095, Jiangsu, Peoples R China
基金
中国国家自然科学基金;
关键词
Ochratoxin A; Tight junction; Reactive oxygen species; Intracellular calcium level; Myosin light chain kinase; BRAIN ENDOTHELIAL-CELLS; OXIDATIVE STRESS; OCHRATOXIN-A; LIQUID-CHROMATOGRAPHY; IN-VITRO; EXPOSURE; EXPRESSION; MYCOTOXIN; KINASE; PHOSPHORYLATION;
D O I
10.1016/j.envpol.2018.06.062
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
Ochratoxin A (OTA) is a frequent contaminant of feed and food worldwide. The toxicity of OTA on intestinal barrier was investigated in porcine intestinal epithelial cells (IPEC-J2). We observed that OTA induced intestinal barrier dysfunction as indicated by the reduction in transepithelial electrical resistance (TEER) and elevation in paracellular permeability to 4 kDa dextran. The barrier dysfunction was accompanied with tight junction disruption including a down-regulation in ZO-1 expression and redistribution of Occludin and ZO-1. Moreover, OTA exposure increased reactive oxygen species (ROS) generation, elevated the intracellular calcium level (Ca2+](c)) and activated myosin light chain kinase (MLCK). Simultaneously, NAC, a ROS scavenger, blocked OTA-induced ROS generation, [Ca2+](c) elevation, barrier dysfunction and tight junction disruption, suggesting that OTA-induced ROS generation may act as a trigger. Next, we found that OTA-induced MLCK activation was inhibited by BAPTA-AM, the cytosolic Ca2+ chelator, demonstrating that OTA-induced MLCK activation is dependent on [Ca2+](c) elevation. Furthermore, inhibition of MLCK with ML-7 or inhibition of [Ca2+](c) elevation with BAPTA-AM markedly prevented OTA-induced barrier dysfunction and tight junction disruption. Taken together, our results indicated that OTA induces ROS generation, and then elevates the [Ca2+](c) and MLCK activity in turn, which finally induces barrier dysfunction and disrupts tight junction in IPEC-J2 cell monolayers. (C) 2018 Elsevier Ltd. All rights reserved.
引用
收藏
页码:106 / 112
页数:7
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