title: Bacillus amyloliquefaciens SC06 alleviates the oxidative stress of IPEC-1 via modulating Nrf2/Keap1 signaling pathway and decreasing ROS production.
abstract: Oxidative stress (OS) plays a major role in the gastrointestinal disorders. Although probiotics were reported to repress OS, few researches compared the antioxidant ability of different Bacillus strains and deciphered the mechanisms. To select a Bacillus strain with higher antioxidant capacity, we used H2O2to induce intestinal porcine epithelial cell 1 (IPEC-1) OS model. The most suitable H2O2concentration and incubation time were determined by the half lethal dose and methyl thiazolyl tetrazolium. Correlation analysis was performed to choose a sensitive indicator for OS. As for the comparison of Bacillus, cells were divided into control, Bacillus treatment, H2O2treatment, and Bacillus pre-protection + H2O2treatment. Bacillus were co-cultured with IPEC-1 for 3 h in Bacillus and Bacillus pre-protection + H2O2treatments. Then, based on OS model, 300 μmol/L H2O2was added into medium of H2O2and Bacillus pre-protection + H2O2treatments for another 12 h. Antioxidant and apoptosis gene expressions were detected to screen the target strain. Nuclear factor erythroid-derived 2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein1 (Keap1) pathway, reactive oxygen species (ROS) production, mitochondrial membrane potential (Δψm), apoptosis, and necrosis were analyzed. Results revealed that heme oxygenase-1 (HO-1) gene expression had a positive correlation with H2O2induction. Moreover, Bacillus amyloliquefaciens SC06 (SC06)-meditated IPEC-1 showed the best antioxidant capacity though modulating Nrf2 phosphorylation. Δψm was elevated, while ROS generation was reduced with SC06 pre-protection, resulting in decreased apoptosis and necrosis. Altogether, HO-1 expression could be regarded as an OS indicator. The regulation of Nrf2/Keap1 pathway and ROS production by SC06 are involved in alleviating OS of IPEC-1.
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Aikun
authors.initials: A
authors.lastname: Fu
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gong
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China. wfli@zju.edu.cn.
authors.forename: Weifen
authors.initials: W
authors.lastname: Li
authors.affiliation: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China. liyali06@163.com.
authors.forename: Yali
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D000975:Antioxidants; D017209:Apoptosis / Q000235:genetics; D000069977:Bacillus amyloliquefaciens / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology; D000072019:Kelch-Like ECH-Associated Protein 1 / Q000235:genetics / Q000378:metabolism; D050983:NF-E2 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D013552:Swine
chemical_list: D000975:Antioxidants; D003470:Culture Media; D000072019:Kelch-Like ECH-Associated Protein 1; D050983:NF-E2 Transcription Factor; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide
keywords: Bacillus; IPEC-1; NOX; Nrf2; Oxidative stress; ROS