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口腔益生菌乳杆菌刺激口腔上皮样细胞转录谱的综合分析

Comprehensive analysis of transcriptional profiles in oral epithelial-like cells stimulated with oral probiotic Lactobacillus spp.

机构信息

Department of Oral Biology & Engineering, Division of Oral Health Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima, 734-8553, Japan.

Department of Medical System Engineering, Division of Oral Health Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima, 734-8553, Japan.

出版信息

Arch Oral Biol. 2020 Oct;118:104832. doi: 10.1016/j.archoralbio.2020.104832. Epub 2020 Jul 25.

Abstract

OBJECTIVE

The mechanisms of action of probiotics can vary among species and among strains of a single species; thus, they can affect host cells in a complex manner. In the present study, Lactobacillus spp. were evaluated for their ability to adhere to gingival epithelial-like cells. Comprehensive analyses of transcriptional profiles of mouse gingival epithelial GE1 cells treated with L. rhamnosus L8020 were performed to assess the putative in vivo probiotic potential of this strain.

METHODS

Five Lactobacillus spp., isolated from the oral cavity, traditional Bulgarian yoghurt, and the feces of a healthy human, were each co-cultured with GE1 cells. Adhesion assays with serial dilution plating and DNA microarray analysis were performed to identify differentially expressed genes (DEGs) in GE1 cells grown in co-culture with L. rhamnosus L8020.

RESULTS

The oral isolates L. rhamnosus L8020, L. casei YU3, and L. paracasei YU4 demonstrated significantly greater adhesion compared with the non-oral isolates. In total, 536 genes in GE1 cells exhibited more than twofold upregulation or downregulation, compared with the 0 h timepoint, during co-culture with L. rhamnosus L8020. Gene ontology enrichment analysis revealed that DEGs were differentially enriched in a time-dependent manner. Early responses involved widespread changes in gene expression.

CONCLUSIONS

This study reveals changes in expression of genes involved in the epithelial physical barrier and immune response in gingival epithelial-like cells co-cultured with L. rhamnosus L8020. Further investigations regarding the molecular mechanisms by which L. rhamnosus L8020 serves as a probiotic may provide evidence to support clinical use.

摘要

目的

不同种属的益生菌作用机制不同,同种属不同菌株的作用机制也不同;因此,它们可以以复杂的方式影响宿主细胞。本研究评估了乳杆菌属对牙龈上皮样细胞的黏附能力。对经鼠牙龈上皮 GE1 细胞用鼠李糖乳杆菌 L8020 处理后的转录谱进行全面分析,以评估该菌株潜在的体内益生菌作用。

方法

将从口腔、传统保加利亚酸奶和健康人粪便中分离出的 5 株乳杆菌属与 GE1 细胞分别共培养。通过连续稀释平板和 DNA 微阵列分析进行黏附实验,以鉴定与 L. rhamnosus L8020 共培养的 GE1 细胞中差异表达的基因(DEGs)。

结果

口腔分离株乳杆菌属 L. rhamnosus L8020、L. casei YU3 和 L. paracasei YU4 的黏附能力明显强于非口腔分离株。与 0 h 相比,在与 L. rhamnosus L8020 共培养时,GE1 细胞中有 536 个基因的表达上调或下调超过两倍。基因本体论富集分析显示,DEGs 以时间依赖的方式差异富集。早期反应涉及基因表达的广泛变化。

结论

本研究揭示了与 L. rhamnosus L8020 共培养的牙龈上皮样细胞中与上皮物理屏障和免疫反应相关的基因表达变化。对 L. rhamnosus L8020 作为益生菌的分子机制的进一步研究可能为临床应用提供证据支持。

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