Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells

The widespread misuse of antibiotics in livestock production has raised growing concerns about antimicrobial resistance and residue contamination. These challenges have led to global restrictions on the use of antibiotic growth promoters (AGPs). However, the ban on AGPs has made the management of in...

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Main Authors: Shiyu Chen, Tiannian Hu, Le Xu, Jiqin Li, Chen Liu, Bangquan Zeng, Qiuye Lin, Zhenhui Cao
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1573479/full
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author Shiyu Chen
Tiannian Hu
Le Xu
Le Xu
Jiqin Li
Chen Liu
Bangquan Zeng
Qiuye Lin
Zhenhui Cao
Zhenhui Cao
author_facet Shiyu Chen
Tiannian Hu
Le Xu
Le Xu
Jiqin Li
Chen Liu
Bangquan Zeng
Qiuye Lin
Zhenhui Cao
Zhenhui Cao
author_sort Shiyu Chen
collection DOAJ
description The widespread misuse of antibiotics in livestock production has raised growing concerns about antimicrobial resistance and residue contamination. These challenges have led to global restrictions on the use of antibiotic growth promoters (AGPs). However, the ban on AGPs has made the management of intestinal inflammation significantly more difficult, highlighting the urgent need for safe and effective alternatives. Lactic acid bacteria (LAB), known for their immunomodulatory properties, have emerged as promising candidates, though their anti-inflammatory mechanisms remain poorly understood. This study investigated the anti-inflammatory effects and underlying molecular mechanisms of a porcine-derived Lactobacillus reuteri (L. reuteri) strain, SBC5-3. Using tumor necrosis factor-alpha (TNF-α)-induced inflammatory models in the HT-29 cells, we employed RNA sequencing (RNA-seq) combined with Western blot analyses to systematically explore the modulation of inflammatory signaling pathways. RNA-seq demonstrated that L. reuteri SBC5-3 downregulated 9 out of 14 TNF-α-induced genes associated with the TNF and/or nuclear factor kappa B (NF-κB) signaling pathways, including BIRC3, PTGS2, CCL20, TNFAIP3, LTB, CXCL1, CXCL10, IL-8, and CSF1. Specifically, L. reuteri SBC5-3 downregulated key genes in the NF-κB pathway, such as TAK1, IKKα, and NFKB1, while upregulating NFKBIA. Concurrent transcriptional suppression occurred in mitogen-activated protein kinase (MAPK) signaling pathway components ERK and JNK. Western blot analysis further confirmed attenuation of phosphorylation involving TAK1, IKKα/β, and IκBα following SBC5-3 treatment. The collective data demonstrate that immunomodulation mediated by L. reuteri SBC5-3 involves inhibiting IκBα degradation, which is known to be essential for the nuclear translocation of the p50/p65 heterodimer, thereby suggesting inhibition of TNF-α-induced NF-κB nuclear translocation. These results position L. reuteri SBC5-3 as a viable therapeutic agent for inflammation modulation through targeted intervention in NF-κB and MAPK signaling pathways.
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spelling doaj-art-c993e4aed0fc4cc688baa4edac5f803f2025-08-20T02:43:39ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.15734791573479Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cellsShiyu Chen0Tiannian Hu1Le Xu2Le Xu3Jiqin Li4Chen Liu5Bangquan Zeng6Qiuye Lin7Zhenhui Cao8Zhenhui Cao9Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Kunming, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, ChinaYunnan Center for Animal Disease Control and Prevention, Kunming, ChinaSchool of Ethnic Medicine, Yunnan Minzu University, Kunming, ChinaFaculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Kunming, ChinaThe widespread misuse of antibiotics in livestock production has raised growing concerns about antimicrobial resistance and residue contamination. These challenges have led to global restrictions on the use of antibiotic growth promoters (AGPs). However, the ban on AGPs has made the management of intestinal inflammation significantly more difficult, highlighting the urgent need for safe and effective alternatives. Lactic acid bacteria (LAB), known for their immunomodulatory properties, have emerged as promising candidates, though their anti-inflammatory mechanisms remain poorly understood. This study investigated the anti-inflammatory effects and underlying molecular mechanisms of a porcine-derived Lactobacillus reuteri (L. reuteri) strain, SBC5-3. Using tumor necrosis factor-alpha (TNF-α)-induced inflammatory models in the HT-29 cells, we employed RNA sequencing (RNA-seq) combined with Western blot analyses to systematically explore the modulation of inflammatory signaling pathways. RNA-seq demonstrated that L. reuteri SBC5-3 downregulated 9 out of 14 TNF-α-induced genes associated with the TNF and/or nuclear factor kappa B (NF-κB) signaling pathways, including BIRC3, PTGS2, CCL20, TNFAIP3, LTB, CXCL1, CXCL10, IL-8, and CSF1. Specifically, L. reuteri SBC5-3 downregulated key genes in the NF-κB pathway, such as TAK1, IKKα, and NFKB1, while upregulating NFKBIA. Concurrent transcriptional suppression occurred in mitogen-activated protein kinase (MAPK) signaling pathway components ERK and JNK. Western blot analysis further confirmed attenuation of phosphorylation involving TAK1, IKKα/β, and IκBα following SBC5-3 treatment. The collective data demonstrate that immunomodulation mediated by L. reuteri SBC5-3 involves inhibiting IκBα degradation, which is known to be essential for the nuclear translocation of the p50/p65 heterodimer, thereby suggesting inhibition of TNF-α-induced NF-κB nuclear translocation. These results position L. reuteri SBC5-3 as a viable therapeutic agent for inflammation modulation through targeted intervention in NF-κB and MAPK signaling pathways.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1573479/fullLactobacillus reuterinuclear factor-κB signalingtumor necrosis factor-alphaanti-inflammatoryintestinal epithelial cells
spellingShingle Shiyu Chen
Tiannian Hu
Le Xu
Le Xu
Jiqin Li
Chen Liu
Bangquan Zeng
Qiuye Lin
Zhenhui Cao
Zhenhui Cao
Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells
Frontiers in Microbiology
Lactobacillus reuteri
nuclear factor-κB signaling
tumor necrosis factor-alpha
anti-inflammatory
intestinal epithelial cells
title Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells
title_full Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells
title_fullStr Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells
title_full_unstemmed Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells
title_short Lactobacillus reuteri SBC5-3 suppresses TNF-α-induced inflammatory responses via NF-κB pathway inhibition in intestinal epithelial cells
title_sort lactobacillus reuteri sbc5 3 suppresses tnf α induced inflammatory responses via nf κb pathway inhibition in intestinal epithelial cells
topic Lactobacillus reuteri
nuclear factor-κB signaling
tumor necrosis factor-alpha
anti-inflammatory
intestinal epithelial cells
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1573479/full
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