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潜在益生菌鼠李糖乳杆菌(MTCC-5897)通过调节宿主紧密连接基因反应抑制大肠杆菌损伤的肠道屏障功能。

Potential Probiotic Lactobacillus rhamnosus (MTCC-5897) Inhibits Escherichia coli Impaired Intestinal Barrier Function by Modulating the Host Tight Junction Gene Response.

机构信息

Animal Biochemistry Division, ICAR-National Dairy Research Institute, Karnal, Haryana, 132001, India.

出版信息

Probiotics Antimicrob Proteins. 2020 Sep;12(3):1149-1160. doi: 10.1007/s12602-019-09608-8.

Abstract

Probiotic as a preventive medicine is emerging as an indispensable tool in addressing the foodborne infections or gastrointestinal disorders. The present study was sought to determine the in vitro prophylactic potential of probiotic Lactobacillus rhamnosus (LR: MTCC-5897) against Escherichia coli (ATCC 14948) induced impairment in intestinal barrier function using Caco-2 cells. Intestinal cells exposed to E. coli demonstrated significantly higher phenol red flux (p < 0.05) and concomitantly decreased TEER (0.69 ± 0.01) in contrast to control or L. rhamnosus (10 cfu/mL)-treated cells. However, E. coli-induced barrier hyperpermeability was restored to significant extents (p < 0.01) when E. coli were excluded, competed or displaced by probiotic LR. Similarly, exposure of Caco-2 cells to E. coli reduced the mRNA expression of key tight junction genes, viz. Zo-1, Claudin-1, Occludin and Cingulin which however were restored significantly (p < 0.05) with L. rhamnosus treatment during exclusion or competition than displacement assays. The protective behaviour of probiotic LR against E. coli can also be observed in immunofluorescent and electron micrograph where intact cellular morphology along with preserved distribution and localisation of key integrity proteins can be found in LR-treated cells in contrast to distorted and disorganised distribution observed with E. coli exposure. In conclusion, L. rhamnosus inhibited and re-established E. coli-impaired intestinal barrier function by improving the expression and distribution of key junction protein and hence could serve an essential food additive to address the various health complications especially those associated with gastrointestinal tract.

摘要

益生菌作为一种预防医学,正在成为解决食源性感染或胃肠道疾病的不可或缺的工具。本研究旨在确定益生菌鼠李糖乳杆菌(LR:MTCC-5897)对大肠杆菌(ATCC 14948)诱导的肠道屏障功能损伤的体外预防潜力,使用 Caco-2 细胞。与对照或鼠李糖乳杆菌(10 cfu/mL)处理的细胞相比,暴露于大肠杆菌的肠细胞显示出明显更高的苯酚红通量(p < 0.05),同时伴随跨上皮电阻(TEER)降低(0.69 ± 0.01)。然而,当排除、竞争或取代益生菌 LR 时,大肠杆菌诱导的屏障通透性增加可显著恢复(p < 0.01)。同样,暴露于大肠杆菌的 Caco-2 细胞减少了关键紧密连接基因的 mRNA 表达,即 Zo-1、Claudin-1、Occludin 和 Cingulin,然而,在用鼠李糖乳杆菌处理时,通过排除或竞争测定比取代测定可显著恢复(p < 0.05)。益生菌 LR 对大肠杆菌的保护作用也可以在免疫荧光和电子显微镜中观察到,在用 LR 处理的细胞中可以发现完整的细胞形态以及关键完整性蛋白的保留分布和定位,而在暴露于大肠杆菌的细胞中则可以观察到扭曲和无序的分布。总之,鼠李糖乳杆菌通过改善关键连接蛋白的表达和分布抑制和重建了大肠杆菌损伤的肠道屏障功能,因此可以作为一种重要的食品添加剂,用于解决各种健康并发症,特别是与胃肠道相关的并发症。

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