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On-line Access: 2023-06-13

Received: 2022-12-27

Revision Accepted: 2023-03-01

Crosschecked: 2023-07-21

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 ORCID:

Mingliang JIN

https://orcid.org/0000-0001-6358-5307

Yizhen WANG

https://orcid.org/0000-0002-3908-7102

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Journal of Zhejiang University SCIENCE B 2023 Vol.24 No.6 P.496-509

http://doi.org/10.1631/jzus.B2200674


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


Author(s):  Chaoyue WEN, Hong ZHANG, Qiuping GUO, Yehui DUAN, Sisi CHEN, Mengmeng HAN, Fengna LI, Mingliang JIN, Yizhen WANG

Affiliation(s):  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; more

Corresponding email(s):   yzwang321@zju.edu.cn, mljin@zju.edu.cn

Key Words:  Engineered probiotics, Intestine, Oxidative injury, Weaned piglets, Nuclear factor-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) pathway


Chaoyue WEN, Hong ZHANG, Qiuping GUO, Yehui DUAN, Sisi CHEN, Mengmeng HAN, Fengna LI, Mingliang JIN, Yizhen WANG. Engineered Bacillus subtilis alleviates intestinal oxidative injury through Nrf2-Keap1 pathway in enterotoxigenic Escherichia coli (ETEC) K88-infected piglet[J]. Journal of Zhejiang University Science B, 2023, 24(6): 496-509.

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author="Chaoyue WEN, Hong ZHANG, Qiuping GUO, Yehui DUAN, Sisi CHEN, Mengmeng HAN, Fengna LI, Mingliang JIN, Yizhen WANG",
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pages="496-509",
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publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2200674"
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%A Chaoyue WEN
%A Hong ZHANG
%A Qiuping GUO
%A Yehui DUAN
%A Sisi CHEN
%A Mengmeng HAN
%A Fengna LI
%A Mingliang JIN
%A Yizhen WANG
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A1 - Chaoyue WEN
A1 - Hong ZHANG
A1 - Qiuping GUO
A1 - Yehui DUAN
A1 - Sisi CHEN
A1 - Mengmeng HAN
A1 - Fengna LI
A1 - Mingliang JIN
A1 - Yizhen WANG
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B2200674


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

文超越1,张红1,郭秋平2,段叶辉2,陈思思2,3,韩萌萌2,3,李凤娜2,靳明亮1,汪以真1
1饲料科学研究所, 浙江省动物饲料与营养重点实验室,教育部分子动物营养重点实验室, 农业部华东动物营养与饲料重点实验室, 浙江大学动物科学学院,中国杭州市,310058
2动物营养生理与代谢过程湖南省重点实验室, 中国科学院亚热带农业生态过程重点实验室, 畜禽养殖污染控制与资源化利用国家工程实验室, 中国科学院亚热带农业生态研究所,中国长沙市,410125
3中国科学院大学现代农业科学学院, 中国北京市,100039
摘要: 工程益生菌具有产生重组免疫刺激物质的特性,可以作为一种治疗药物。本研究使用基因工程技术构建了表达抗菌肽KR32的重组枯草芽孢杆菌(WB800-KR32),并且探究了其在通过激活Nrf2-Keap1途径对产肠毒素大肠埃希氏菌(ETEC) K88感染断奶仔猪导致的肠道氧化态紊乱的保护作用。我们将28头断奶仔猪随机分成4组,每组7个重复,均饲喂基础日粮。对照组灌喂灭菌生理盐水;ETEC组、ETEC+WB800组和ETEC+WB800-KR32组分别在第1~14天灌喂灭菌生理盐水、5×1010 CFU WB800、5×1010 CFU WB800-KR32,在第15-17天灌喂ETEC K88 1×1010 CFU。结果表明,WB800-KR32预处理能够缓解ETEC K88导致的肠道紊乱,提高肠道粘膜抗氧化酶活性(过氧化物酶、超氧化物歧化酶和谷胱甘肽过氧化物酶),降低丙二醛含量。更重要的是,WB800-KR32预处理可上调回肠粘膜Nrf2的蛋白表达量,同时下调Keap1的蛋白表达量。此外,WB800-KR32预处理还显著改变了粪便微生物的丰度(Ace和Chao指数),并增加了Eubacterium_rectale_ ATCC_33656在粪便中的丰度。综上,WB800-KR32可能通过Nrf2-Keap1途径缓解ETEC K88导致的肠道氧化损伤,这为将WB800-KR32作为调节ETEC K88感染导致的肠道氧化失调的潜在治疗手段提供了一个新的视角。

关键词:工程益生菌;肠道;氧化损伤;断奶仔猪;Nrf2-Keap1途径

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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