Similar articles

Abstract: 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, 5×10¹⁰ CFU (CFU: colony forming units) WB800, and 5×10¹⁰ CFU WB800-KR32, respectively, on Days 1‍‒‍14 and all infused with ETEC K88 1×10¹⁰ 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.

工程枯草芽孢杆菌通过Nrf2-Keap1途径缓解ETEC K88感染仔猪导致的肠道氧化损伤

[1]AyabeT, SatchellDP, WilsonCL, et al., 2000. Secretion of microbicidal α‍-‍defensins by intestinal Paneth cells in response to bacteria. Nat Immunol, 1(2):‍113-118.

[2]BäckhedF, FraserCM, RingelY, et al., 2012. Defining a healthy human gut microbiome: current concepts, future directions, and clinical applications. Cell Host Microbe, 12(5):611-622.

[3]ChuFF, EsworthyRS, DoroshowJH, 2004. Role of Se-dependent glutathione peroxidases in gastrointestinal inflammation and cancer. Free Radic Biol Med, 36(12):1481-1495.

[4]CruzKCP, EnekeghoLO, StuartDT, 2022. Bioengineered probiotics: synthetic biology can provide live cell therapeutics for the treatment of foodborne diseases. Front Bioeng Biotechnol, 10:890479.

[5]DuanYH, ZengLM, LiFN, et al., 2017. Effect of branched-chain amino acid ratio on the proliferation, differentiation, and expression levels of key regulators involved in protein metabolism of myocytes. Nutrition, 36:8-16.

[6]EsworthyRS, SwiderekKM, HoYS, et al., 1998. Selenium-dependent glutathione peroxidase-GI is a major glutathione peroxidase activity in the mucosal epithelium of rodent intestine. Biochim Biophys Acta, 1381(2):213-226.

[7]FanPX, LiuP, SongPX, et al., 2017. Moderate dietary protein restriction alters the composition of gut microbiota and improves ileal barrier function in adult pig model. Sci Rep, 7:43412.

[8]FlorianS, WinglerK, SchmehlK, et al., 2001. Cellular and subcellular localization of gastrointestinal glutathione peroxidase in normal and malignant human intestinal tissue. Free Radic Res, 35(6):655-663.

[9]GóthL, RassP, PáyA, 2004. Catalase enzyme mutations and their association with diseases. Mol Diagn, 8(3):‍141-149.

[10]GuanGP, DingSJ, YinYL, et al., 2019. Macleaya cordata extract alleviated oxidative stress and altered innate immune response in mice challenged with enterotoxigenic Escherichia coli. Sci China Life Sci, 62(8):1019-1027.

[11]GuilloteauP, ZabielskiR, HammonHM, et al., 2010. Nutritional programming of gastrointestinal tract development. Is the pig a good model for man? Nutr Res Rev, 23(1):4-22.

[12]GuoPT, ZhangK, MaX, et al., 2020. Clostridium species as probiotics: potentials and challenges. J Anim Sci Biotechnol, 11:24.

[13]HuWY, YangYY, LiZ, et al., 2019. Antibacterial, cytotoxicity and mechanism of the antimicrobial peptide KR-32 in weaning piglets. Int J Pept Res Ther, 26(2):943-953.

[14]HuangJJ, BaiYM, XieWT, et al., 2023. Lycium barbarum polysaccharides ameliorate canine acute liver injury by reducing oxidative stress, protecting mitochondrial function, and regulating metabolic pathways. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 24:157-171.

[15]IghodaroOM, AkinloyeOA, 2018. First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): their fundamental role in the entire antioxidant defence grid. Alex J Med, 54(4):287-293.

[16]JinML, ZhangH, ZhaoK, et al., 2018. Responses of intestinal mucosal barrier functions of rats to simulated weightlessness. Front Physiol, 9:729.

[17]LekshmiM, AmminiP, KumarS, et al., 2017. The food production environment and the development of antimicrobial resistance in human pathogens of animal origin. Microorganisms, 5(1):11.

[18]LiFN, DuanYH, LiYH, et al., 2015. Effects of dietary n-6:n-3 PUFA ratio on fatty acid composition, free amino acid profile and gene expression of transporters in finishing pigs. Br J Nutr, 113(5):739-748.

[19]LiH, MaLT, LiZQ, et al., 2021. Evolution of the gut microbiota and its fermentation characteristics of Ningxiang pigs at the young stage. Animals (Basel), 11(3):638.

[20]LiWF, ZhouXX, LuP, 2004. Bottlenecks in the expression and secretion of heterologous proteins in Bacillus subtilis. Res Microbiol, 155(8):605-610.

[21]LiuH, WangJ, HeT, et al., 2018. Butyrate: a double-edged sword for health? Adv Nutr, 9(1):21-29.

[22]LiuHY, CaoXX, WangH, et al., 2019. Antimicrobial peptide KR-32 alleviates Escherichia coli K88-induced fatty acid malabsorption by improving expression of fatty acid transporter protein 4 (FATP4). J Anim Sci, 97(6):2342-2356.

[23]LiuSN, ZhangB, XiangDC, et al., 2021. Effect of Pediococcus pentosaceus 368 on grow performance, fecal microbiota and metabolite in pigs. Microbiol China, 48(6):2035-2048 (in Chinese).

[24]LuanC, ZhangHW, SongDG, et al., 2014a. Expressing antimicrobial peptide cathelicidin-BF in Bacillus subtilis using SUMO technology. Appl Microbiol Biotechnol, 98(8):3651-3658.

[25]LuanC, XieYG, PuYT, et al., 2014b. Recombinant expression of antimicrobial peptides using a novel self-cleaving aggregation tag in Escherichia coli. Can J Microbiol, 60(3):113-120.

[26]LuiseD, LauridsenC, BosiP, et al., 2019. Methodology and application of Escherichia coli F4 and F18 encoding infection models in post-weaning pigs. J Anim Sci Biotechnol, 10:53.

[27]LyakhovichVV, VavilinVA, ZenkovNK, et al., 2006. Active defense under oxidative stress. The antioxidant responsive element. Biochemistry (Mosc), 71(9):962-974.

[28]NandiA, YanLJ, JanaCK, et al., 2019. Role of catalase in oxidative stress- and age-associated degenerative diseases. Oxid Med Cell Longev, 2019:9613090.

[29]National Research Council, 2012. Nutrients Requirements of Swine, 11th Ed. National Academy Press, Washington, USA, p.20-26.

[30]RajputSA, LiangSJ, WangXQ, et al., 2021. Lycopene protects intestinal epithelium from deoxynivalenol-induced oxidative damage via regulating Keap1/Nrf2 signaling. Antioxidants, 10(9):1493.

[31]RenM, CaiS, ZhouT, et al., 2019. Isoleucine attenuates infection induced by E. coli challenge through the modulation of intestinal endogenous antimicrobial peptide expression and the inhibition of the increase in plasma endotoxin and IL-6 in weaned pigs. Food Funct, 10(6):3535-3542.

[32]RiviereA, GagnonM, WeckxS, et al., 2015. Mutual cross-feeding interactions between Bifidobacterium longum subsp. longum NCC2705 and Eubacterium rectale ATCC 33656 explain the bifidogenic and butyrogenic effects of arabinoxylan oligosaccharides. Appl Environ Microbiol, 81(22):7767-7781.

[33]RouraE, KoopmansSJ, LallesJP, et al., 2016. Critical review evaluating the pig as a model for human nutritional physiology. Nutr Res Rev, 29(1):60-90.

[34]ShiY, HuY, WangZQ, et al., 2022. The Protective effect of taurine on oxidized fish-oil-induced liver oxidative stress and intestinal barrier-function impairment in juvenile Ictalurus punctatus. Antioxidants, 10(11):1690.

[35]SmithF, ClarkJE, OvermanBL, et al., 2010. Early weaning stress impairs development of mucosal barrier function in the porcine intestine. Am J Physiol Gastrointest Liver Physiol, 298(3):G352-G363.

[36]TangYL, LiFN, TanB, et al., 2014. Enterotoxigenic Escherichia coli infection induces intestinal epithelial cell autophagy. Vet Microbiol, 171(1-2):160-164.

[37]TsikasD, 2017. Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: analytical and biological challenges. Anal Biochem, 524:13-30.

[38]WangJ, SuLQ, ZhangL, et al., 2022. Spirulina platensis aqueous extracts ameliorate colonic mucosal damage and modulate gut microbiota disorder in mice with ulcerative colitis by inhibiting inflammation and oxidative stress. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(6):481-501.

[39]WangRJ, LiuN, YangYC, et al., 2021. Flavor supplementation during late gestation and lactation periods increases the reproductive performance and alters fecal microbiota of the sows. Anim Nutr, 7(3):679-687.

[40]WenCY, LiFN, DuanYH, et al., 2019. Dietary taurine regulates free amino acid profiles and taurine metabolism in piglets with diquat-induced oxidative stress. J Funct Foods, 62:103569.

[41]WenCY, LiFN, GuoQP, et al., 2020a. Protective effects of taurine against muscle damage induced by diquat in 35 days weaned piglets. J Anim Sci Biotechnol, 11:56.

[42]WenCY, GuoQP, WangWL, et al., 2020b. Taurine alleviates intestinal injury by mediating tight junction barriers in diquat-challenged piglet models. Front Physiol, 11:449.

[43]WenCY, LiSY, WangJJ, et al., 2021. Heat stress alters the intestinal microbiota and metabolomic profiles in mice. Front Microbiol, 12:706772.

[44]WierupM, 2001. The Swedish experience of the 1986 year ban of antimicrobial growth promoters, with special reference to animal health, disease prevention, productivity, and usage of antimicrobials. Microb Drug Resist, 7(2):183-190.

[45]WuT, ShiYT, ZhangYY, et al., 2021. Lactobacillus rhamnosus LB1 alleviates enterotoxigenic Escherichia coli-induced adverse effects in piglets by improving host immune response and anti-oxidation stress and restoring intestinal integrity. Front Cell Infect Microbiol, 11:724401.

[46]WuX, ZhangY, LiuZ, et al., 2012. Effects of oral supplementation with glutamate or combination of glutamate and N-carbamylglutamate on intestinal mucosa morphology and epithelium cell proliferation in weanling piglets. J Anim Sci, 90(S4):337-339.

[47]XiaXJ, ZhangXL, LiuMC, et al., 2021. Toward improved human health: efficacy of dietary selenium on immunity at the cellular level. Food Funct, 12(3):976-989.

[48]XiaYY, BinP, LiuSJ, et al., 2018. Enterotoxigenic Escherichia coli infection promotes apoptosis in piglets. Microb Pathog, 125:290-294.

[49]XiaYY, ChenSY, ZhaoYY, et al., 2019. GABA attenuates ETEC-induced intestinal epithelial cell apoptosis involving GABA_AR signaling and the AMPK-autophagy pathway. Food Funct, 10(11):7509-7522.

[50]XieWC, SongLY, WangX, et al., 2021. A bovine lactoferricin-lactoferrampin-encoding Lactobacillus reuteri CO21 regulates the intestinal mucosal immunity and enhances the protection of piglets against enterotoxigenic Escherichia coli K88 challenge. Gut Microbes, 13(1):1956281.

[51]XiongW, HuangJ, LiXY, et al., 2020. Icariin and its phosphorylated derivatives alleviate intestinal epithelial barrier disruption caused by enterotoxigenic Escherichia coli through modulate p38 MAPK in vivo and in vitro. FASEB J, 34(1):1783-1801.

[52]YangB, YueY, ChenY, et al., 2021. Lactobacillus plantarum CCFM1143 alleviates chronic diarrhea via inflammation regulation and gut microbiota modulation: a double-blind, randomized, placebo-controlled study. Front Immunol, 12:746585.

[53]YangWY, ChouCH, WangC, 2022. The effects of feed supplementing Akkemansia muciniphila on incidence, severity, and gut microbiota of necrotic enteritis in chickens. Poult Sci, 101(4):101751.

[54]YinJ, WuMM, XiaoH, et al., 2014. Development of an antioxidant system after early weaning in piglets. J Anim Sci, 92(2):612-619.

[55]YounisNS, AbduldaiumMS, MohamedME, 2020. Protective effect of geraniol on oxidative, inflammatory and apoptotic alterations in isoproterenol-induced cardiotoxicity: role of the Keap1/Nrf2/HO-1 and PI3K/Akt/mTOR pathways. Antioxidants, 9(10):977.

[56]YuE, ChenDW, YuB, et al., 2021. Amelioration of enterotoxigenic Escherichia coli-induced disruption of intestinal epithelium by manno-oligosaccharide in weaned pigs. J Funct Foods, 82:104492.

[57]ZhangQS, WidmerG, TziporiS, 2013. A pig model of the human gastrointestinal tract. Gut Microbes, 4(3):193-200.

[58]ZhouJ, XiongX, YinJ, et al., 2019. Dietary lysozyme alters sow’s gut microbiota, serum immunity and milk metabolite profile. Front Microbiol, 10:177.

[59]ZongX, FuJ, XuBC, et al., 2020. Interplay between gut microbiota and antimicrobial peptides. Anim Nutr, 6(4):389-396.