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猪源罗伊氏乳杆菌的体外评价:益生菌特性及其对产肠毒素大肠杆菌K88攻击的猪肠道上皮细胞的影响

In Vitro Evaluation of Swine-Derived Lactobacillus reuteri: Probiotic Properties and Effects on Intestinal Porcine Epithelial Cells Challenged with Enterotoxigenic Escherichia coli K88.

作者信息

Wang Zhilin, Wang Li, Chen Zhuang, Ma Xianyong, Yang Xuefen, Zhang Jian, Jiang Zongyong

机构信息

State Key Laboratory of Livestock and Poultry Breeding, Ministry of Agriculture Key Laboratory of Animal Nutrition and Feed Science in South China, Guangdong Public Laboratory of Animal Breeding and Nutrition, Guangdong Key Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong 510640, P.R. China.

出版信息

J Microbiol Biotechnol. 2016 Jun 28;26(6):1018-25. doi: 10.4014/jmb.1510.10089.

Abstract

Probiotics are considered as the best effective alternatives to antibiotics. The aim of this study was to characterize the probiotic potential of lactobacilli for use in swine farming by using in vitro evaluation methods. A total of 106 lactic acid bacterial isolates, originating from porcine feces, were first screened for the capacity to survive stresses considered important for putative probiotic strains. Sixteen isolates showed notable acid and bile resistance, antibacterial activity, and adherence to intestinal porcine epithelial cells (IPEC-1). One isolate, LR1, identified as Lactobacillus reuteri, was selected for extensive study of its probiotic and functional properties in IPEC-1 cell models. L. reuteri LR1 exhibited good adhesion to IPEC-1 cells and could inhibit the adhesion of enterotoxigenic Escherichia coli (ETEC) to IPEC-1 cells. L. reuteri LR1 could also modulate transcript and protein expression of cytokines involved in inflammation in IPEC-1 cells; the Lactobacillus strain inhibited the ETEC-induced expression of proinflammatory transcripts (IL-6 and TNF-α) and protein (IL-6), and increased the level of anti-inflammatory cytokine (IL-10). Measurement of the permeation of FD-4 showed that L. reuteri LR1 could maintain barrier integrity in monolayer IPEC-1 cells exposed to ETEC. Immunolocalization experiments showed L. reuteri LR1 could also prevent ETEC-induced tight junction ZO-1 disruption. Together, these results indicate that L. reuteri LR1 exhibits desirable probiotic properties and could be a potential probiotic for use in swine production.

摘要

益生菌被认为是抗生素的最佳有效替代品。本研究的目的是通过体外评估方法来表征用于养猪业的乳酸菌的益生菌潜力。首先对总共106株源自猪粪便的乳酸菌分离株进行筛选,以检测其在对假定益生菌菌株而言重要的应激条件下的存活能力。16株分离株表现出显著的耐酸和耐胆汁能力、抗菌活性以及对猪肠上皮细胞(IPEC-1)的黏附性。其中一株名为LR1的分离株被鉴定为罗伊氏乳杆菌,被选用于在IPEC-1细胞模型中对其益生菌和功能特性进行深入研究。罗伊氏乳杆菌LR1对IPEC-1细胞表现出良好的黏附性,并且能够抑制产肠毒素大肠杆菌(ETEC)对IPEC-1细胞的黏附。罗伊氏乳杆菌LR1还可以调节IPEC-1细胞中参与炎症的细胞因子的转录和蛋白质表达;该乳杆菌菌株抑制了ETEC诱导的促炎转录本(IL-6和TNF-α)和蛋白质(IL-6)的表达,并提高了抗炎细胞因子(IL-10)的水平。FD-4通透性测量结果表明,罗伊氏乳杆菌LR1能够在暴露于ETEC的单层IPEC-1细胞中维持屏障完整性。免疫定位实验表明,罗伊氏乳杆菌LR1还可以防止ETEC诱导的紧密连接ZO-1破坏。总之,这些结果表明罗伊氏乳杆菌LR1具有理想的益生菌特性,可能是一种可用于养猪生产的潜在益生菌。

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