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乳杆菌益生菌调节 LPS 刺激的树突状细胞的抗菌反应基因转录。

Lactobacilli Probiotics Modulate Antibacterial Response Gene Transcription of Dendritic Cells Challenged with LPS.

机构信息

Restorative Dentistry Department, Federal University of Piauí, Campus Universitário Ministro Petrônio Portella, Bairro Ininga, CEP: 64049-550, Teresina, Brazil.

Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.

出版信息

Probiotics Antimicrob Proteins. 2024 Feb;16(1):293-307. doi: 10.1007/s12602-023-10043-z. Epub 2023 Jan 25.

Abstract

Probiotics are beneficial bacteria that may modulate the immune response by altering the maturation and function of antigen-presenting cells, such as dendritic cells. This study aimed to evaluate the antibacterial gene expression of dendritic cells challenged with LPS and probiotics. Immature dendritic cells were obtained from human CD14 monocytes and challenged with E. coli LPS and probiotics Lacticaseibacillus rhamnosus (LR-32) and Lactobacillus acidophilus (LA-5) at a ratio DC:bacteria of 1:10. The analysis of gene expression was performed by RT-qPCR using the Kit RT2 human antibacterial response. In the supernatant, the cytokines secretion was determined by ELISA. Tukey post-ANOVA with p at 5% was used for statistical analysis. LPS showed the higher upregulation of 29 genes compared with the groups where probiotics were added to LPS, including genes related to an inflammatory response like BIRC3, CASP1, CCL5, CXCL1, IL12B, IL18, MYD88, NLRP3, RIPK1, and TIRAP. Similarly, LPS increased the transcription of genes enrolled with apoptosis such as CARD6, CASP1, IRF5, MAP2K1, MAP2K4, MAPK1, MYD88, NLRP3, RIPK2, TNF, TNFRSF1A, and XIAP when compared to probiotics groups (p < 0.05). Although probiotics decrease several genes upregulated by LPS, the transcription of encoded cytokines IL12A, IL12B, IL1B, IL6, CXCL8, and TNF genes was maintained upregulated by probiotics, except for IL18, which was downregulated by LA-5. LA-5 led to a higher transcription of IL1B, IL6, and CXCL-8 which was followed by the secretion of these proteins by ELISA. The results suggest that probiotics attenuate the transcription of inflammatory and immune response genes caused by LPS.

摘要

益生菌是有益细菌,通过改变树突状细胞等抗原呈递细胞的成熟和功能,可能调节免疫反应。本研究旨在评估 LPS 和益生菌刺激的树突状细胞的抗菌基因表达。从人 CD14 单核细胞中获得未成熟的树突状细胞,并以 1:10 的 DC:细菌比例用大肠杆菌 LPS 和益生菌鼠李糖乳杆菌(LR-32)和嗜酸乳杆菌(LA-5)刺激。使用 RT2 人类抗菌反应试剂盒通过 RT-qPCR 进行基因表达分析。在细胞培养上清液中,通过 ELISA 测定细胞因子分泌。采用 Tukey 事后方差分析(p 值为 5%)进行统计分析。与添加益生菌的 LPS 组相比,LPS 显示出 29 个基因的更高上调,包括与炎症反应相关的基因,如 BIRC3、CASP1、CCL5、CXCL1、IL12B、IL18、MYD88、NLRP3、RIPK1 和 TIRAP。同样,与益生菌组相比,LPS 增加了凋亡相关基因的转录,如 CARD6、CASP1、IRF5、MAP2K1、MAP2K4、MAPK1、MYD88、NLRP3、RIPK2、TNF、TNFRSF1A 和 XIAP(p<0.05)。尽管益生菌降低了 LPS 上调的几个基因的转录,但编码细胞因子 IL12A、IL12B、IL1B、IL6、CXCL8 和 TNF 的转录仍被益生菌上调,除了 LA-5 下调的 IL18。LA-5 导致 IL1B、IL6 和 CXCL-8 的转录更高,随后通过 ELISA 分泌这些蛋白质。结果表明,益生菌可减弱 LPS 引起的炎症和免疫反应基因的转录。

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