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益生菌和后生元干预对临床耐甲氧西林金黄色葡萄球菌(MRSA)具有抗黏附作用,并可阻止MRSA诱导的HT-29细胞系肠道上皮细胞高通透性。

Probiotic and postbiotic interference exhibit anti-adhesion effects against clinical methicillin-resistant Staphylococcus aureus (MRSA) and impede MRSA-induced intestinal epithelial hyper-permeability in HT-29 cell line.

作者信息

Nataraj Basavaprabhu Haranahalli, Ramesh Chette, Mallappa Rashmi Hogarehalli

机构信息

Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India; Dairy Bacteriology Section, Southern Regional Station, ICAR-National Dairy Research Institute, Adugodi, 560030, Bengaluru, Karnataka, India.

Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India.

出版信息

Microb Pathog. 2025 Feb;199:107215. doi: 10.1016/j.micpath.2024.107215. Epub 2024 Dec 6.

Abstract

This study investigates the dynamics of MRSA de-colonization on HT-29 cell line using effective strategies like probiotics and postbiotics. Exploring novel alternatives to combat infections caused by antibiotic-resistant pathogens is an urgent need. Harnessing the antagonistic properties of live probiotics and their heat-killed preparations (postbiotics) to curb the growth of AMR pathogens represents a promising and essential area of contemporary research. This study was designed to evaluate the anti-adhesion properties of indigenous probiotics (Limosilactobacillus fermentum Lf1 and Lactiplantibacillus plantarum A5), as well as standard reference strains (Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus NCFM), and their heat-killed postbiotic preparations against clinical MRSA isolates (MRSA12/206 and 5/255) on the HT-29 cell line. ATR-FTIR-based functional group characterization of the postbiotic preparations revealed the heat-induced alterations in cell surface molecules and architecture. Both probiotic and postbiotic preparations were non-cytotoxic to HT-29 cells. The probiotic intervention, via protective, competitive, and displacement modes, significantly (p < 0.05) reduced the adhesion of MRSA isolates to HT-29 cells, with the protective and competitive modes showing greater efficacy. In contrast, heat-killed probiotics demonstrated notable anti-MRSA adhesion effects across all three modes (protective, competitive, and displacement). In comparison, heat-killed cells exhibited a superior anti-adhesion capability compared to live cells, likely due to the enhanced accessibility of microbe-associated molecular patterns and adhesion sites following heat treatment. Furthermore, co-treatment of MRSA with probiotic strains substantially (p < 0.05) reduced FITC-dextran transflux across the HT-29 cell monolayer. In conclusion, this study highlights the superior anti-adhesion efficacy of heat-killed postbiotics over live probiotic cells against MRSA isolates. It underscores the further need for pre-clinical and in-vivo investigations to validate the anti-MRSA colonization and gut barrier prophylactic or therapeutic potential of the investigated probiotics and postbiotics. Thus, the present study documents and supports the alternative to antibiotics potential of probiotics and postbiotics.

摘要

本研究利用益生菌和后生元等有效策略,研究了耐甲氧西林金黄色葡萄球菌(MRSA)在HT-29细胞系上的去定植动态。探索对抗抗生素耐药病原体引起感染的新方法迫在眉睫。利用活益生菌及其热灭活制剂(后生元)的拮抗特性来抑制AMR病原体的生长,是当代研究中一个有前景且重要的领域。本研究旨在评估本土益生菌(发酵乳杆菌Lf1和植物乳杆菌A5)以及标准参考菌株(鼠李糖乳杆菌GG和嗜酸乳杆菌NCFM)及其热灭活后生元制剂对HT-29细胞系上临床MRSA分离株(MRSA12/206和5/255)的抗黏附特性。基于衰减全反射傅里叶变换红外光谱(ATR-FTIR)的后生元制剂官能团表征揭示了热诱导的细胞表面分子和结构变化。益生菌和后生元制剂对HT-29细胞均无细胞毒性。益生菌干预通过保护、竞争和取代模式,显著(p<0.05)降低了MRSA分离株对HT-29细胞的黏附,其中保护和竞争模式显示出更高的功效。相比之下,热灭活益生菌在所有三种模式(保护、竞争和取代)中均表现出显著的抗MRSA黏附作用。相比之下,热灭活细胞比活细胞表现出更强的抗黏附能力,这可能是由于热处理后微生物相关分子模式和黏附位点的可及性增强。此外,MRSA与益生菌菌株共同处理显著(p<0.05)降低了FITC-葡聚糖通过HT-29细胞单层的通量。总之,本研究强调了热灭活后生元对MRSA分离株的抗黏附功效优于活益生菌细胞。它强调了进一步进行临床前和体内研究以验证所研究的益生菌和后生元的抗MRSA定植及肠道屏障预防或治疗潜力的必要性。因此,本研究记录并支持了益生菌和后生元作为抗生素替代品的潜力。

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