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利用RNA测序分析鉴定在富含益生菌乳酸杆菌属的小鼠脾脏中表达改变的免疫相关基因。

Identification of immune-associated genes with altered expression in the spleen of mice enriched with probiotic Lactobacillus species using RNA-seq profiling.

作者信息

Truong Anh Duc, Tran Ha Thi Thanh, Chu Nhu Thi, Phan Lanh, Phan Hoai Thi, Dang Thu Huong, Dang Hoang Vu, Nguyen La Anh

机构信息

Department of Biochemistry and Immunology, National Institute of Veterinary Research, Dong Da, Hanoi 100000, Vietnam.

Department of Microbial Biotechnology, Food Industries Research Institute, Thanh Xuan Distr., Hanoi 100000, Vietnam.

出版信息

Anim Biosci. 2025 Feb;38(2):336-349. doi: 10.5713/ab.24.0280. Epub 2024 Aug 26.

Abstract

OBJECTIVE

Probiotics are living microorganisms that can provide health benefits when consumed. Here, we investigated the effects of probiotics on gene expression in the spleen of mice using RNA-sequencing analysis between negative control and probiotic groups (including 4 Lactobacillus strains: Lactobacillus fermentum, L. casei, L. plantarum, and L. brevis).

METHODS

Mice exposed with probiotic in 4 weeks by intragastric administration. Then, spleen tissues of the control and probiotics groups were collected on days 14 and 28 for RNA sequencing.

RESULTS

In total, 665, 186, and 81 differentially expressed genes (DEGs) were significantly expressed on day 14 vs control, day 28 vs control groups, and probiotics day 28 vs day 14 groups, respectively. On the other hand, 12 toll-like receptor genes underwent additional validation through quantitative real-time polymerase chain reaction (qRT-PCR), affirming the increased alignment between qRT-PCR and RNA-Seq findings. In addition, the Kyoto encyclopedia of genes and genomes and gene ontology analyses revealed that the DEGs were predominantly enriched in defense responses to pathogens, including inflammatory bowel diseases, malaria, leukaemia virus 1, and herpes virus, as well as immune processes related to immune response and signal transduction. This study represents the first investigation into mice's gene expression in the spleen exposed to probiotics using Lactobacillus spp. isolated from a field strain in Vietnam.

CONCLUSION

Our results provide valuable insights into the impacts and functions of probiotics on mammalian development, offering crucial information for the potential therapeutic use of probiotics in defending against pathogens in Vietnam. The findings from this study highlight the potential of probiotics in modulating gene expression in the spleen, which may have implications for immune function and overall health in mice.

摘要

目的

益生菌是一类活的微生物,食用后可对健康有益。在此,我们通过对阴性对照组和益生菌组(包括4种乳酸杆菌菌株:发酵乳杆菌、干酪乳杆菌、植物乳杆菌和短乳杆菌)进行RNA测序分析,研究了益生菌对小鼠脾脏基因表达的影响。

方法

通过胃内给药对小鼠进行4周的益生菌暴露处理。然后,在第14天和第28天收集对照组和益生菌组的脾脏组织进行RNA测序。

结果

分别在第14天与对照组、第28天与对照组以及益生菌组第28天与第14天相比,总共发现665个、186个和81个差异表达基因(DEG)显著表达。另一方面,通过定量实时聚合酶链反应(qRT-PCR)对12个Toll样受体基因进行了额外验证,证实了qRT-PCR与RNA测序结果之间的一致性增强。此外,京都基因与基因组百科全书(KEGG)和基因本体(GO)分析表明,差异表达基因主要富集在对病原体的防御反应中,包括炎症性肠病、疟疾、白血病病毒1和疱疹病毒,以及与免疫反应和信号转导相关的免疫过程。本研究首次对从越南田间菌株分离的乳酸杆菌属益生菌暴露后小鼠脾脏中的基因表达进行了研究。

结论

我们的结果为益生菌对哺乳动物发育的影响和功能提供了有价值的见解,为益生菌在越南抵御病原体的潜在治疗应用提供了关键信息。本研究结果突出了益生菌在调节小鼠脾脏基因表达方面的潜力,这可能对小鼠的免疫功能和整体健康产生影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3d1/11725755/bf4a9b635b4d/ab-24-0280f1.jpg

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