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葡萄糖通过影响不耐热肠毒素的产生,显著增强产肠毒素大肠杆菌对肠道上皮细胞的黏附。

Glucose significantly enhances enterotoxigenic Escherichia coli adherence to intestinal epithelial cells through its effects on heat-labile enterotoxin production.

作者信息

Wijemanne Prageeth, Moxley Rodney A

机构信息

School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, United States of America.

出版信息

PLoS One. 2014 Nov 19;9(11):e113230. doi: 10.1371/journal.pone.0113230. eCollection 2014.

Abstract

The present study tested whether exposure of enterotoxigenic Escherichia coli (ETEC) to glucose at different concentrations in the media results in increased bacterial adherence to host cells through increased heat-labile enterotoxin (LT) production, thereby suggesting the effects are physiological. Porcine-origin ETEC strains grown in Casamino acid yeast extract medium containing different concentrations of glucose were washed and inoculated onto IPEC-J2 porcine intestinal epithelial cells to test for effects on adherence and host cell cAMP concentrations. Consistent with previous studies, all LT+ strains had higher ETEC adherence to IPEC-J2 cells than did LT- strains. Adherence of the LT- but not the LT+ strains was increased by pre-incubating the IPEC-J2 cells with LT and decreased by co-incubation with GM1 ganglioside in a dose-dependent manner (P<0.05). To determine whether the glucose concentration of the cell culture media has an effect on adherence, IPEC-J2 cells were inoculated with LT+ or LT- strains in cell culture media containing a final glucose concentration of 0, 0.25, 0.5, 1.0 or 2.0%, and incubated for 4 h. Only media containing 0.25% glucose resulted in increased adherence and cAMP levels, and this was limited to IPEC-J2 cells inoculated with LT+ strains. This study supports the hypothesis that glucose, at a concentration optimal for LT expression, enhances bacterial adherence through the promotion of LT production. Hence, these results establish the physiological relevance of the effects of glucose on LT production and provide a basis for how glucose intake may influence the severity of ETEC infection.

摘要

本研究检测了产肠毒素大肠杆菌(ETEC)在培养基中暴露于不同浓度葡萄糖时,是否会通过增加不耐热肠毒素(LT)的产生而导致细菌对宿主细胞的黏附增加,从而表明这些影响具有生理学意义。将在含有不同浓度葡萄糖的酪蛋白氨基酸酵母提取物培养基中培养的猪源ETEC菌株洗涤后接种到IPEC-J2猪肠道上皮细胞上,以检测对黏附和宿主细胞cAMP浓度的影响。与先前的研究一致,所有LT+菌株对IPEC-J2细胞的ETEC黏附均高于LT-菌株。用LT预孵育IPEC-J2细胞可增加LT-菌株(而非LT+菌株)的黏附,且与GM1神经节苷脂共孵育可使其黏附呈剂量依赖性降低(P<0.05)。为了确定细胞培养基中的葡萄糖浓度是否对黏附有影响,将LT+或LT-菌株接种到最终葡萄糖浓度为0、0.25、0.5、1.0或2.0%的细胞培养基中的IPEC-J2细胞上,并孵育4小时。只有含有0.25%葡萄糖的培养基导致黏附和cAMP水平增加,且这仅限于接种LT+菌株的IPEC-J2细胞。本研究支持以下假设:葡萄糖在最适合LT表达的浓度下,通过促进LT的产生增强细菌黏附。因此,这些结果确立了葡萄糖对LT产生影响的生理学相关性,并为葡萄糖摄入如何影响ETEC感染的严重程度提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1823/4237375/ecb3063712fb/pone.0113230.g001.jpg

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