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Engineered Bacillus subtilis alleviates intestinal oxidative injury through Nrf2-Keap1 pathway in enterotoxigenic Escherichia coli(ETEC) K88-infected piglet

查看全文 作　　者：Chaoyue [1,2,3]WEN;Hong [1]ZHANG;Qiuping [1]GUO;Yehui [1]DUAN;Sisi [1]CHEN;Mengmeng [1]HAN;Fengna [1]LI;Mingliang [1]JIN;Yizhen [1]WANG 高影响力作者 机构地区：[1]Institute of Feed Science,Key Laboratory of Animal Feed and Nutrition of Zhejiang Province,Key Laboratory of Molecular Animal Nutrition,Ministry of Education,Key Laboratory of Animal Nutrition and Feed Science in Eastern China,Ministry of Agriculture,College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;[2]Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process,CAS Key Laboratory of Agro-Ecological Processes in Subtropical Region,National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production,Institute of Subtropical Agriculture,Chinese Academy of Sciences,Changsha 410125,China;[3]College of Advanced Agricultural Science,University of Chinese Academy of Sciences,Bejing 100039,China高影响力机构 出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》索引2023年第24卷第6期,共14页高影响力期刊 基　　金：supported by the Zhejiang Provincial Key R&D Program of China(No.2021C02008);the China Agriculture Research System of MOF and MARA(No.CARS-35);the National Natural Science Foundation of China(No.32022079);the Fundamental Research Funds for the Central Universities(No.2022QZJH46);the Taishan Industrial Leading Talents Project. 摘　　要：Engineered probiotics can serve as therapeutics based on their ability of produce recombinant immune-stimulating properties.In this study,we built the recombinant Bacillus subtilis WB800 expressing antimicrobial peptide KR32(WB800-KR32)using genetic engineering methods and investigated its protective effects of nuclear factor-E2-related factor 2(Nrf2)-Kelch-like ECH-associated protein 1(Keap1)pathway activation in intestinal oxidative disturbance induced by enterotoxigenic Escherichia coli(ETEC)K88 in weaned piglets.Twenty-eight weaned piglets were randomly distributed into four treatment groups with seven replicates fed with a basal diet.The feed of the control group(CON)was infused with normal sterilized saline;meanwhile,the ETEC,ETEC+WB800,and ETEC+WB800-KR32 groups were orally administered normal sterilized saline,5×10^(10)CFU(CFU:colony forming units)WB800,and 5×10^(10)CFU WB800-KR32,respectively,on Days 1-14 and all infused with ETEC K881×10^(10)CFU on Days 15-17.The results showed that pretreatment with WB800-KR32 attenuated ETEC-induced intestinal disturbance,improved the mucosal activity of antioxidant enzyme(catalase(CAT),superoxide dismutase(SOD),and glutathione peroxidase(GPx))and decreased the content of malondialdehyde(MDA).More importantly,WB800-KR32 downregulated genes involved in antioxidant defense(GPx and SOD1).Interestingly,WB800-KR32 upregulated the protein expression of Nrf2 and downregulated the protein expression of Keap1 in the ileum.WB800-KR32 markedly changed the richness estimators(Ace and Chao)of gut microbiota and increased the abundance of Eubacterium_rectale_ATCC_33656 in the feces.The results suggested that WB800-KR32 may alleviate ETEC-induced intestinal oxidative injury through the Nrf2-Keap1 pathway,providing a new perspective for WB800-KR32 as potential therapeutics to regulate intestinal oxidative disturbance in ETEC K88 infection. 关 键 词：Engineered probiotics Intestine Oxidative injury Weaned piglets Nuclear factor-E2-related factor 2(Nrf2)-Kelch-like ECH-associated protein 1(Keap1)pathway
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