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马奶酒乳杆菌通过调节紧密连接改善小鼠腹泻。

Lactobacillus paracasei from koumiss ameliorates diarrhea in mice via tight junctions modulation.

机构信息

College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China.

College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.

出版信息

Nutrition. 2022 Jun;98:111584. doi: 10.1016/j.nut.2021.111584. Epub 2022 Jan 19.

Abstract

OBJECTIVES

Probiotics are gaining interest as alternative options for antibiotic or antiinflammatory drugs. Probiotics can affect the health of the host through metabolites and competitive inhibition adhesion of pathogenic microorganisms. Koumiss is an important part of the diet of Asian nomads, and is rich in a broad array of probiotics that can benefit the body. Mongolians have developed koumiss therapy to assist in the treatment of various diseases. In the present study, we investigate the beneficial effect of Lactobacillus paracasei, a strain isolated from koumiss, on a mouse model of diarrhea induced by Escherichia coli O (E. coli O).

METHODS

Probiotics were isolated from Mongolian koumiss. The resistance of probiotics against acid, bile salts, gastric juice, and intestinal juice was evaluated. The mouse model of diarrhea was established by the intragastric administration of E. coli O after NaHCO treatment. L. paracasei was intragastrically administered before or after E. coli O exposure in mice. The plasma levels of diamine oxidase and zounlin were quantified using an enzyme-linked immunosorbent assay, and the integrity of the intestinal barrier and goblet cells of mice with diarrhea were observed using hematoxylin and eosin and Alcian blue periodic acid-Schiff staining. The expression of tight junction (TJ) proteins was detected by immunohistochemistry and Western blot.

RESULTS

A total of five lactic acid bacteria and two yeast strains were isolated from koumiss, and L. paracasei was screened for animal experiments. Experimental results showed that L. paracasei could reduce the increase in diamine oxidase and zonulin caused by E. coli (P < 0.05); increase goblet cells and the expression of TJ proteins ZO-1, occludin, and claudin-1 (P < 0.05); increase the expression of mucin 2, oligomeric mucus/gel-forming (P < 0.05) protein; and reduce the level of inhibitor kappa B-alpha and myosin light-chain kinase.

CONCLUSIONS

L. paracasei reduced the intestinal permeability, induced the expression of mucin 2, oligomeric mucus/gel-forming protein, and increased the number of goblet cells in mice by the upregulation of the expression of TJ proteins via the nuclear factor kappa B cells-myosin light-chain kinase signaling pathway.

摘要

目的

益生菌作为抗生素或抗炎药物的替代选择越来越受到关注。益生菌可以通过代谢物和竞争性抑制致病微生物的黏附来影响宿主的健康。马奶酒是亚洲游牧民族饮食的重要组成部分,富含多种益生菌,对人体有益。蒙古人已经开发出马奶酒疗法来辅助治疗各种疾病。本研究旨在探讨从马奶酒中分离出的副干酪乳杆菌对大肠杆菌 O(E. coli O)诱导的腹泻小鼠模型的有益作用。

方法

从蒙古马奶酒中分离益生菌。评估益生菌对酸、胆盐、胃液和肠液的抵抗力。用 NaHCO3 处理后,通过灌胃给予大肠杆菌 O 建立腹泻小鼠模型。在暴露于大肠杆菌 O 之前或之后,将副干酪乳杆菌通过灌胃给予小鼠。用酶联免疫吸附试验定量测定血浆中二胺氧化酶和 ZOUNLIN 的水平,用苏木精和伊红及阿尔辛蓝过碘酸希夫染色观察腹泻小鼠的肠屏障完整性和杯状细胞。用免疫组织化学和 Western blot 检测紧密连接(TJ)蛋白的表达。

结果

从马奶酒中分离出 5 株乳酸菌和 2 株酵母菌,筛选出副干酪乳杆菌进行动物实验。实验结果表明,副干酪乳杆菌可降低大肠杆菌引起的二胺氧化酶和 ZOUNLIN 增加(P<0.05);增加杯状细胞和 TJ 蛋白 ZO-1、occludin 和 claudin-1 的表达(P<0.05);增加黏蛋白 2、寡聚黏液/凝胶形成(P<0.05)蛋白的表达;并降低抑制因子 kappa B-alpha 和肌球蛋白轻链激酶的水平。

结论

副干酪乳杆菌通过核因子 kappa B 细胞-肌球蛋白轻链激酶信号通路上调 TJ 蛋白的表达,降低肠道通透性,诱导黏蛋白 2、寡聚黏液/凝胶形成蛋白的表达,增加 TJ 蛋白的表达,增加杯状细胞数量。

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