Bacillus amyloliquefaciens SC06 alleviates the oxidative stress of IPEC-1 via modulating Nrf2/Keap1

• 作 者: Yang Wang Yanping Wu Yibing Wang Aiktn Fu Li Gong Weifen Li Yali Li
• 机 构: Key Laboratory of Molecular Animal Nutrition of the Ministry of Education, Institute of Feed Science
• 出 处: 中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会
• 发表日期: 2016年12期
• 关键词: Bacillus IPEC-1 Oxidative stress Nrt2 ROS NOX

　　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 decipher the mechanisms.To select a Bacillus strain with higher antioxidant capacity, we used H2O2 to induce intestinal porcine epithelial cell 1 (IPEC-1) OS model.The most suitable H2O2 concentration 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, H2O2 treatment and Bacillus pre-protection+H2O2 treatment.Bacillus were co-cultured with IPEC-1 for 3h in Bacillus and Bacillus pre-protection+H2O2 treatments.Then, based on OS model, 300 μmol/L H2O2 were added into medium of H2O2 and Bacillus pre-protection+H2O2 treatments for another 12h.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 proteinl (Keapl) pathway,reactive oxygen species (ROS) production, mitochondrial membrane potential (△ψm), apoptosis and necrosis were analyzed.Results revealed that heme oxygenase-1 (HO-1) gene expression had a positive correlation with H2O2 induction.Moreover, Bacillus amyloliquefaciens SC06 (SC06)-meditated IPEC-1 showed the best antioxidant capacity though modulating Nrf2 phosphorylation.△ψm 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.