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Probiotic and anti-inflammatory attributes of an isolate Lactobacillus helveticus NS8 from Mongolian fermented koumiss.

Rong J, Zheng H, Liu M, Hu X, Wang T, Zhang X, Jin F, Wang L - BMC Microbiol. (2015)

Bottom Line: NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis.Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10.In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: The Affiliated Hospital, School of Medicine, Hangzhou Normal University, Hangzhou, China. jjrong@hznu.edu.cn.

ABSTRACT

Background: Koumiss is a traditionally fermented mare's milk described with health-promoting potentials for decades. However, only a few studies focused on the probiotic strains isolated from koumiss. In this study, we collected koumiss samples from Inner Mongolian pasturing area of China and selected a promising strain of Lactobacillus helveticus, isolate NS8, based on the survival abilities in gastrointestinal tract (GIT) and adhesion to intestinal endothelial cells in vitro. As the ability to positively modulate host immune response is a feature of increasing importance in measuring the probiotic potential of a bacterial strain, our study mainly focus on the immunomodulatory properties of L. helveticus NS8 by using in vivo and ex vivo analyses.

Results: L. helveticus NS8 was identified by molecular-typing methods, both at genus and species levels. As a typical food niche-specific bacteria, NS8 showed a moderate survival ability in GIT environment in vitro. However, an excellent binding capacity to the human intestinal epithelial cells, along with significant autoaggregation and cell-surface hydrophobicity was observed. Additionally, the presence of S-layer protein was responsible for the cell surface properties of this strain. NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis. In the meantime, co-culture with NS8 induced an increased level of secretion of anti-inflammatory cytokine IL-10 in peripheral blood mono-nuclear cells (PBMCs). Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10. Specially, adding of the purified S-layer protein didn't influence the production of IL-10. The specific ligand-host receptor interactions on the NS8 specific immune responses need to be learned further.

Conclusion: In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

No MeSH data available.


Related in: MedlinePlus

Modulation of the proinflammatory response triggered by LPS in RAW264.7 mouse macrophages. RAW264.7 cells were stimulated with L. helveticus NS8 (bacteria: cell ratio of 100:1) or its S-layer protein (10 μg/ml), with or without co-incubation of 1 μg/mL LPS for 4 h. Expression profiles of IL10 (a), IL-12p70 (b), and TNF-α (c) were examined by quantitative real-time PCR, indicated as the relative levels to the induction level of the control, which was set at a value of 1. Results are the means ± SD of representative of three independent experiments. Significant differences indicated as: **, P < 0.01; *, P < 0.05
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Fig5: Modulation of the proinflammatory response triggered by LPS in RAW264.7 mouse macrophages. RAW264.7 cells were stimulated with L. helveticus NS8 (bacteria: cell ratio of 100:1) or its S-layer protein (10 μg/ml), with or without co-incubation of 1 μg/mL LPS for 4 h. Expression profiles of IL10 (a), IL-12p70 (b), and TNF-α (c) were examined by quantitative real-time PCR, indicated as the relative levels to the induction level of the control, which was set at a value of 1. Results are the means ± SD of representative of three independent experiments. Significant differences indicated as: **, P < 0.01; *, P < 0.05

Mentions: In a final set of experiments, we tested the effects of NS8 on LPS-evoked inflammatory responses by analyzing the gene expression of two proinflammatory factors through qPCR, TNF-α and IL-12, and the anti-inflammatory cytokine IL-10 in mouse macrophage cell line RAW264.7. Here we also investigated whether the S-layer protein is involved in immunomodulatory activity exerted by NS8. After 4 h of co-stimulation of the RAW264.7 cells with LPS, NS8 induced a pronounced anti-inflammatory profile, as evidenced by an dramatically enhanced induction of IL-10 compared to the induction of TNF-α and IL-12 (Fig. 5). NS8 also decreased LPS-induced proinflammatory IL-12 production significantly (Fig. 5b). The reduction of IL-12 induced by purified S-layer protein was similar to that induced by NS8 strain itself, however, the expression of anti-inflammatory cytokine IL-10 was not influenced by S-layer protein (Fig. 5a, b). The results obtained by employing LPS-induced macrophages, therefore, confirmed the anti-inflammatory immune responses of NS8, but not dependent on S-layer protein.Fig. 5


Probiotic and anti-inflammatory attributes of an isolate Lactobacillus helveticus NS8 from Mongolian fermented koumiss.

Rong J, Zheng H, Liu M, Hu X, Wang T, Zhang X, Jin F, Wang L - BMC Microbiol. (2015)

Modulation of the proinflammatory response triggered by LPS in RAW264.7 mouse macrophages. RAW264.7 cells were stimulated with L. helveticus NS8 (bacteria: cell ratio of 100:1) or its S-layer protein (10 μg/ml), with or without co-incubation of 1 μg/mL LPS for 4 h. Expression profiles of IL10 (a), IL-12p70 (b), and TNF-α (c) were examined by quantitative real-time PCR, indicated as the relative levels to the induction level of the control, which was set at a value of 1. Results are the means ± SD of representative of three independent experiments. Significant differences indicated as: **, P < 0.01; *, P < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4591576&req=5

Fig5: Modulation of the proinflammatory response triggered by LPS in RAW264.7 mouse macrophages. RAW264.7 cells were stimulated with L. helveticus NS8 (bacteria: cell ratio of 100:1) or its S-layer protein (10 μg/ml), with or without co-incubation of 1 μg/mL LPS for 4 h. Expression profiles of IL10 (a), IL-12p70 (b), and TNF-α (c) were examined by quantitative real-time PCR, indicated as the relative levels to the induction level of the control, which was set at a value of 1. Results are the means ± SD of representative of three independent experiments. Significant differences indicated as: **, P < 0.01; *, P < 0.05
Mentions: In a final set of experiments, we tested the effects of NS8 on LPS-evoked inflammatory responses by analyzing the gene expression of two proinflammatory factors through qPCR, TNF-α and IL-12, and the anti-inflammatory cytokine IL-10 in mouse macrophage cell line RAW264.7. Here we also investigated whether the S-layer protein is involved in immunomodulatory activity exerted by NS8. After 4 h of co-stimulation of the RAW264.7 cells with LPS, NS8 induced a pronounced anti-inflammatory profile, as evidenced by an dramatically enhanced induction of IL-10 compared to the induction of TNF-α and IL-12 (Fig. 5). NS8 also decreased LPS-induced proinflammatory IL-12 production significantly (Fig. 5b). The reduction of IL-12 induced by purified S-layer protein was similar to that induced by NS8 strain itself, however, the expression of anti-inflammatory cytokine IL-10 was not influenced by S-layer protein (Fig. 5a, b). The results obtained by employing LPS-induced macrophages, therefore, confirmed the anti-inflammatory immune responses of NS8, but not dependent on S-layer protein.Fig. 5

Bottom Line: NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis.Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10.In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: The Affiliated Hospital, School of Medicine, Hangzhou Normal University, Hangzhou, China. jjrong@hznu.edu.cn.

ABSTRACT

Background: Koumiss is a traditionally fermented mare's milk described with health-promoting potentials for decades. However, only a few studies focused on the probiotic strains isolated from koumiss. In this study, we collected koumiss samples from Inner Mongolian pasturing area of China and selected a promising strain of Lactobacillus helveticus, isolate NS8, based on the survival abilities in gastrointestinal tract (GIT) and adhesion to intestinal endothelial cells in vitro. As the ability to positively modulate host immune response is a feature of increasing importance in measuring the probiotic potential of a bacterial strain, our study mainly focus on the immunomodulatory properties of L. helveticus NS8 by using in vivo and ex vivo analyses.

Results: L. helveticus NS8 was identified by molecular-typing methods, both at genus and species levels. As a typical food niche-specific bacteria, NS8 showed a moderate survival ability in GIT environment in vitro. However, an excellent binding capacity to the human intestinal epithelial cells, along with significant autoaggregation and cell-surface hydrophobicity was observed. Additionally, the presence of S-layer protein was responsible for the cell surface properties of this strain. NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis. In the meantime, co-culture with NS8 induced an increased level of secretion of anti-inflammatory cytokine IL-10 in peripheral blood mono-nuclear cells (PBMCs). Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10. Specially, adding of the purified S-layer protein didn't influence the production of IL-10. The specific ligand-host receptor interactions on the NS8 specific immune responses need to be learned further.

Conclusion: In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

No MeSH data available.


Related in: MedlinePlus