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长双歧杆菌HY8001对大肠杆菌O157:H7志贺毒素的抑制活性。

Inhibitory activity of Bifidobacterium longum HY8001 against Vero cytotoxin of Escherichia coli O157:H7.

作者信息

Kim S H, Yang S J, Koo H C, Bae W K, Kim J Y, Park J H, Baek Y J, Park Y H

机构信息

Department of Microbiology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Suwon, Gyunggi, Korea.

出版信息

J Food Prot. 2001 Nov;64(11):1667-73. doi: 10.4315/0362-028x-64.11.1667.

Abstract

Vero cytotoxin (VT)-producing Escherichia coli (VTEC), such as E. coli O157:H7, are emerging foodborne pathogens worldwide. VTs are associated with hemorrhagic colitis and hemolytic uremic syndrome in humans. Attachment of the B subunit of VTs to its receptor, globotriaosylceramide (Gb3), at gut epithelium is the primary step and, consequently, the A subunit of VTs inhibits protein synthesis in the target cell. Proinflammatory cytokines, such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, up-regulate Gb3 expression, increase sensitivity to VTs, and enhance VT action in developing disease. Currently, there is a growing interest in probiotics, given the increasing occurrence of antibiotic-resistant bacteria. In particular, much work on bifidobacteria among probiotics, regarded as microorganisms targeted for technological and therapeutic applications, has been performed. In Korea, the neutralizing effect of the culture supernatant of Bifidobacterium longum HY8001, Korean isolate, against the VTs from E. coli O157:H7 was found. Therefore, this study focused on the raveling of the inhibitory effect of B. longum HY8001 against VTs, through the interference B subunit of VTs and Gb3 interaction. Mice were inoculated intragastrically with B. longum HY8001 culture supernatant before and after challenge with E. coli O157:H7. Control mice were inoculated intragastrically only with E. coli O157:H7. Cytokine, TNF-alpha, and IL-1beta levels in sera and expression of their mRNA were decreased, and expression of Gb3 in renal tubular epithelial cells was reduced in mice treated with B. longum HY8001 culture supernatant. In competitive enzyme-linked immunosorbent assays (ELISAs), the culture supernatant of B. longum HY8001 primarily binds VTs to interfere the VTs with Gb3 interaction. These results suggest that soluble substance(s) in B. longum HY8001 culture supernatant may have inhibitory activity on the expression of Gb3, VT-Gb3 interaction, or both. Further study should be done to elucidate the property of soluble substances in B. longum HY8001 culture supernatant.

摘要

产志贺毒素大肠杆菌(VTEC),如大肠杆菌O157:H7,是全球范围内新出现的食源性病原体。志贺毒素与人类的出血性结肠炎和溶血性尿毒症综合征有关。志贺毒素的B亚基在肠道上皮细胞与它的受体球三糖基神经酰胺(Gb3)结合是第一步,因此,志贺毒素的A亚基抑制靶细胞中的蛋白质合成。促炎细胞因子,如肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β,上调Gb3表达,增加对志贺毒素的敏感性,并增强志贺毒素在疾病发展中的作用。目前,鉴于抗生素耐药菌的出现日益增多,人们对益生菌的兴趣与日俱增。特别是,已经对益生菌中的双歧杆菌进行了大量研究,双歧杆菌被视为具有技术和治疗应用价值的微生物。在韩国,发现了韩国分离株长双歧杆菌HY8001的培养上清液对大肠杆菌O157:H7产生的志贺毒素具有中和作用。因此,本研究聚焦于通过干扰志贺毒素的B亚基与Gb3的相互作用来阐明长双歧杆菌HY8001对志贺毒素的抑制作用。在用大肠杆菌O157:H7攻击前后,给小鼠灌胃长双歧杆菌HY8001培养上清液。对照小鼠仅灌胃大肠杆菌O157:H7。在用长双歧杆菌HY8001培养上清液处理的小鼠中,血清中的细胞因子、TNF-α和IL-1β水平及其mRNA表达降低,肾小管上皮细胞中Gb3的表达减少。在竞争性酶联免疫吸附测定(ELISA)中,长双歧杆菌HY8001的培养上清液主要结合志贺毒素,以干扰志贺毒素与Gb3的相互作用。这些结果表明,长双歧杆菌HY8001培养上清液中的可溶性物质可能对Gb3的表达、志贺毒素-Gb3相互作用或两者都具有抑制活性。应进一步研究以阐明长双歧杆菌HY8001培养上清液中可溶性物质的性质。

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