title: Autophagy protects intestinal epithelial cells against deoxynivalenol toxicity by alleviating oxidative stress via IKK signaling pathway.
abstract: Autophagy is an intracellular process of homeostatic degradation that promotes cell survival under various stressors. Deoxynivalenol (DON), a fungal toxin, often causes diarrhea and disturbs the homeostasis of the intestinal system. To investigate the function of intestinal autophagy in response to DON and associated mechanisms, we firstly knocked out ATG5 (autophagy-related gene 5) in porcine intestinal epithelial cells (IPEC-J2) using CRISPR-Cas9 technology. When treated with DON, autophagy was induced in IPEC-J2 cells but not in IPEC-J2.Atg5ko cells. The deficiency in autophagy increased DON-induced apoptosis in IPEC-J2.atg5ko cells, in part, through the generation of reactive oxygen species (ROS). The cellular stress response can be restored in IPEC-J2.atg5ko cells by overexpressing proteins involved in protein folding. Interestingly, we found that autophagy deficiency downregulated the expression of endoplasmic reticulum folding proteins BiP and PDI when IPEC-J2.atg5ko cells were treated with DON. In addition, we investigated the molecular mechanism of autophagy involved in the IKK, AMPK, and mTOR signaling pathway and found that Bay-117082 and Compound C, specific inhibitors for IKK and AMPK, respectively, inhibited the induction of autophagy. Taken together, our results suggest that autophagy is pivotal for protection against DON in pig intestinal cells.
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China. Electronic address: tangyulong@isa.ac.cn.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China. Electronic address: lifengna@isa.ac.cn.
authors.forename: Fengna
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, University of New Mexico School of Medicine, Albuquerque, New Mexico 87131.
authors.forename: Chien-An A
authors.initials: CA
authors.lastname: Hu
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Peng
authors.initials: P
authors.lastname: Liao
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Kunrong
authors.initials: K
authors.lastname: Tan
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Bie
authors.initials: B
authors.lastname: Tan
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Xia
authors.initials: X
authors.lastname: Xiong
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China.
authors.forename: Tiejun
authors.initials: T
authors.lastname: Li
authors.affiliation: Key Laboratory of Agroecology and Processing of Subtropical Region, Institute of Subtropical Agriculture, Research Center for Healthy Breeding of Livestock and Poultry, Hunan Engineering and Research Center for Animal and Poultry Science, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central China, The Chinese Academy of Science, Ministry of Agriculture, 410125 Changsha City, Hunan, People's Republic of China; School of Life Sciences, Hunan Normal University, Changsha 41008, China.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Yin
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D005434:Flow Cytometry; D051550:I-kappa B Kinase / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000187:drug effects / Q000473:pathology*; D018384:Oxidative Stress / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D014255:Trichothecenes / Q000633:toxicity*
chemical_list: D014255:Trichothecenes; D051550:I-kappa B Kinase; C007262:deoxynivalenol
keywords: Apoptosis intestinal homeostasis; Autophagy; Deoxynivalenol; Double nicking Crisper-Cas9