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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

Influence of S-layer protein on the cell surface properties of L. helveticus NS8. a. SDS-PAGE profile with Coomassie blue staining of extracted S-layer protein by LiCl treatment, indicating an approximate mass value of 42 kDa. Lane 1, low molecular weight protein standards; lane 2, 5 μg of purified protein was loaded on the gel. b. Comparison of the autoaggregation ability of NS8 before (●) and after removal of their surface layer proteins (○). Error bars represent standard deviations (SD) of the mean values of results from three replicate experiments. c. Bacterial micrographs of NS8, showing the morphology difference between autoaggregated cells (left) and those cells after removal of S-layer proteins (right). Magnification, × 100
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Fig2: Influence of S-layer protein on the cell surface properties of L. helveticus NS8. a. SDS-PAGE profile with Coomassie blue staining of extracted S-layer protein by LiCl treatment, indicating an approximate mass value of 42 kDa. Lane 1, low molecular weight protein standards; lane 2, 5 μg of purified protein was loaded on the gel. b. Comparison of the autoaggregation ability of NS8 before (●) and after removal of their surface layer proteins (○). Error bars represent standard deviations (SD) of the mean values of results from three replicate experiments. c. Bacterial micrographs of NS8, showing the morphology difference between autoaggregated cells (left) and those cells after removal of S-layer proteins (right). Magnification, × 100

Mentions: For extraction the S-layer protein from NS8 bacteria, a generally employed method for the removal of S-layer proteins from the cell surfaces, LiCl extraction, was applied for NS8. SDS-PAGE revealed the molecular mass of the protein is approximately 42 kDa, basically in line with those characterized S-layer proteins (Fig. 2a). We verified whether the protocol based on LiCl washes efficiently removed most of the S-layer proteins from the surface of L. helveticus cells. After S-layer removal, autoaggregation ability of NS8 reduced dramatically from 70 to 32 % (Fig. 2b). The bacterial morphology also changed from appearance of cluster into separated cells under microscope (Fig. 2c). These results indicated that the presence of S-layer protein contributed to the cell surface properties of NS8.Fig. 2


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)

Influence of S-layer protein on the cell surface properties of L. helveticus NS8. a. SDS-PAGE profile with Coomassie blue staining of extracted S-layer protein by LiCl treatment, indicating an approximate mass value of 42 kDa. Lane 1, low molecular weight protein standards; lane 2, 5 μg of purified protein was loaded on the gel. b. Comparison of the autoaggregation ability of NS8 before (●) and after removal of their surface layer proteins (○). Error bars represent standard deviations (SD) of the mean values of results from three replicate experiments. c. Bacterial micrographs of NS8, showing the morphology difference between autoaggregated cells (left) and those cells after removal of S-layer proteins (right). Magnification, × 100
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig2: Influence of S-layer protein on the cell surface properties of L. helveticus NS8. a. SDS-PAGE profile with Coomassie blue staining of extracted S-layer protein by LiCl treatment, indicating an approximate mass value of 42 kDa. Lane 1, low molecular weight protein standards; lane 2, 5 μg of purified protein was loaded on the gel. b. Comparison of the autoaggregation ability of NS8 before (●) and after removal of their surface layer proteins (○). Error bars represent standard deviations (SD) of the mean values of results from three replicate experiments. c. Bacterial micrographs of NS8, showing the morphology difference between autoaggregated cells (left) and those cells after removal of S-layer proteins (right). Magnification, × 100
Mentions: For extraction the S-layer protein from NS8 bacteria, a generally employed method for the removal of S-layer proteins from the cell surfaces, LiCl extraction, was applied for NS8. SDS-PAGE revealed the molecular mass of the protein is approximately 42 kDa, basically in line with those characterized S-layer proteins (Fig. 2a). We verified whether the protocol based on LiCl washes efficiently removed most of the S-layer proteins from the surface of L. helveticus cells. After S-layer removal, autoaggregation ability of NS8 reduced dramatically from 70 to 32 % (Fig. 2b). The bacterial morphology also changed from appearance of cluster into separated cells under microscope (Fig. 2c). These results indicated that the presence of S-layer protein contributed to the cell surface properties of NS8.Fig. 2

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