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Lactobacillus rhamnosus CNCMI-4317 Modulates Fiaf/Angptl4 in Intestinal Epithelial Cells and Circulating Level in Mice.

Jacouton E, Mach N, Cadiou J, Lapaque N, Clément K, Doré J, van Hylckama Vlieg JE, Smokvina T, Blottière HM - PLoS ONE (2015)

Bottom Line: Lactobacilli are often considered to display beneficial effect for their hosts, acting on different regulatory pathways.We then validated in vivo bacterial effects using C57BL/6 mono-colonized mice fed with normal chow.Moreover, this effect was accompanied by transcriptome modulation of several pathways including immune response and metabolism in vitro.

View Article: PubMed Central - PubMed

Affiliation: Danone Nutricia Research, Palaiseau, France; INRA, UMR 1319 Micalis, Jouy en Josas, France; AgroParistech, UMR Micalis, Jouy en Josas, France.

ABSTRACT

Background and objectives: Identification of new targets for metabolic diseases treatment or prevention is required. In this context, FIAF/ANGPTL4 appears as a crucial regulator of energy homeostasis. Lactobacilli are often considered to display beneficial effect for their hosts, acting on different regulatory pathways. The aim of the present work was to study the effect of several lactobacilli strains on Fiaf gene expression in human intestinal epithelial cells (IECs) and on mice tissues to decipher the underlying mechanisms.

Subjects and methods: Nineteen lactobacilli strains have been tested on HT-29 human intestinal epithelial cells for their ability to regulate Fiaf gene expression by RT-qPCR. In order to determine regulated pathways, we analysed the whole genome transcriptome of IECs. We then validated in vivo bacterial effects using C57BL/6 mono-colonized mice fed with normal chow.

Results: We identified one strain (Lactobacillus rhamnosus CNCMI-4317) that modulated Fiaf expression in IECs. This regulation relied potentially on bacterial surface-exposed molecules and seemed to be PPAR-γ independent but PPAR-α dependent. Transcriptome functional analysis revealed that multiple pathways including cellular function and maintenance, lymphoid tissue structure and development, as well as lipid metabolism were regulated by this strain. The regulation of immune system and lipid and carbohydrate metabolism was also confirmed by overrepresentation of Gene Ontology terms analysis. In vivo, circulating FIAF protein was increased by the strain but this phenomenon was not correlated with modulation Fiaf expression in tissues (except a trend in distal small intestine).

Conclusion: We showed that Lactobacillus rhamnosus CNCMI-4317 induced Fiaf expression in human IECs, and increased circulating FIAF protein level in mice. Moreover, this effect was accompanied by transcriptome modulation of several pathways including immune response and metabolism in vitro.

No MeSH data available.


Related in: MedlinePlus

Bacterial effectors characterization. (a) Conditioned media, (b) Heat inactivated bacteria, (c) Transwell. Cells were incubated 6h with 10% of final volume of bacterial fractions (CM, HI) and 20% of bacteria for transwell structure. Transwell prevented contact between cells and bacteria.Bars represent means of Fiaf relative expression (percentage of rosiglitazone) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene. Stars represent p<0.05 (*), p<0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represents a no significant difference in comparison with negative control.
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pone.0138880.g003: Bacterial effectors characterization. (a) Conditioned media, (b) Heat inactivated bacteria, (c) Transwell. Cells were incubated 6h with 10% of final volume of bacterial fractions (CM, HI) and 20% of bacteria for transwell structure. Transwell prevented contact between cells and bacteria.Bars represent means of Fiaf relative expression (percentage of rosiglitazone) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene. Stars represent p<0.05 (*), p<0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represents a no significant difference in comparison with negative control.

Mentions: To determine the bacterial effector(s) involved in the activation of Fiaf by CNCMI–4317 strain, several bacterial fractions were tested. Conditioned medium (CM) (Fig 3a) and heat inactivated (HI) bacteria (Fig 3b) were not effective on Fiaf up-regulation suggesting that the effector was not a secreted product and was heat sensitive (Fiaf relative expression was: 50±12.53 and 21.33±14.22 respectively for CM and HI vs 92±17.69 and 87.67±4.16 for bacterial strain; P<0.001). The requirement of bacterial-cells direct contact was further assessed. To do so, HT–29 cells were separated from the bacteria using transwellTM permeable support (Corning) in which the bacteria were added (Fig 3c). Using this item, the ability of the lactobacilli to induce Fiaf expression was reduced (33.98±14.03 vs 59.78±19.54) suggesting, at least in part, the requirement of a direct contact of bacterial surface factors with HT–29 cells.


Lactobacillus rhamnosus CNCMI-4317 Modulates Fiaf/Angptl4 in Intestinal Epithelial Cells and Circulating Level in Mice.

Jacouton E, Mach N, Cadiou J, Lapaque N, Clément K, Doré J, van Hylckama Vlieg JE, Smokvina T, Blottière HM - PLoS ONE (2015)

Bacterial effectors characterization. (a) Conditioned media, (b) Heat inactivated bacteria, (c) Transwell. Cells were incubated 6h with 10% of final volume of bacterial fractions (CM, HI) and 20% of bacteria for transwell structure. Transwell prevented contact between cells and bacteria.Bars represent means of Fiaf relative expression (percentage of rosiglitazone) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene. Stars represent p<0.05 (*), p<0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represents a no significant difference in comparison with negative control.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4595210&req=5

pone.0138880.g003: Bacterial effectors characterization. (a) Conditioned media, (b) Heat inactivated bacteria, (c) Transwell. Cells were incubated 6h with 10% of final volume of bacterial fractions (CM, HI) and 20% of bacteria for transwell structure. Transwell prevented contact between cells and bacteria.Bars represent means of Fiaf relative expression (percentage of rosiglitazone) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene. Stars represent p<0.05 (*), p<0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represents a no significant difference in comparison with negative control.
Mentions: To determine the bacterial effector(s) involved in the activation of Fiaf by CNCMI–4317 strain, several bacterial fractions were tested. Conditioned medium (CM) (Fig 3a) and heat inactivated (HI) bacteria (Fig 3b) were not effective on Fiaf up-regulation suggesting that the effector was not a secreted product and was heat sensitive (Fiaf relative expression was: 50±12.53 and 21.33±14.22 respectively for CM and HI vs 92±17.69 and 87.67±4.16 for bacterial strain; P<0.001). The requirement of bacterial-cells direct contact was further assessed. To do so, HT–29 cells were separated from the bacteria using transwellTM permeable support (Corning) in which the bacteria were added (Fig 3c). Using this item, the ability of the lactobacilli to induce Fiaf expression was reduced (33.98±14.03 vs 59.78±19.54) suggesting, at least in part, the requirement of a direct contact of bacterial surface factors with HT–29 cells.

Bottom Line: Lactobacilli are often considered to display beneficial effect for their hosts, acting on different regulatory pathways.We then validated in vivo bacterial effects using C57BL/6 mono-colonized mice fed with normal chow.Moreover, this effect was accompanied by transcriptome modulation of several pathways including immune response and metabolism in vitro.

View Article: PubMed Central - PubMed

Affiliation: Danone Nutricia Research, Palaiseau, France; INRA, UMR 1319 Micalis, Jouy en Josas, France; AgroParistech, UMR Micalis, Jouy en Josas, France.

ABSTRACT

Background and objectives: Identification of new targets for metabolic diseases treatment or prevention is required. In this context, FIAF/ANGPTL4 appears as a crucial regulator of energy homeostasis. Lactobacilli are often considered to display beneficial effect for their hosts, acting on different regulatory pathways. The aim of the present work was to study the effect of several lactobacilli strains on Fiaf gene expression in human intestinal epithelial cells (IECs) and on mice tissues to decipher the underlying mechanisms.

Subjects and methods: Nineteen lactobacilli strains have been tested on HT-29 human intestinal epithelial cells for their ability to regulate Fiaf gene expression by RT-qPCR. In order to determine regulated pathways, we analysed the whole genome transcriptome of IECs. We then validated in vivo bacterial effects using C57BL/6 mono-colonized mice fed with normal chow.

Results: We identified one strain (Lactobacillus rhamnosus CNCMI-4317) that modulated Fiaf expression in IECs. This regulation relied potentially on bacterial surface-exposed molecules and seemed to be PPAR-γ independent but PPAR-α dependent. Transcriptome functional analysis revealed that multiple pathways including cellular function and maintenance, lymphoid tissue structure and development, as well as lipid metabolism were regulated by this strain. The regulation of immune system and lipid and carbohydrate metabolism was also confirmed by overrepresentation of Gene Ontology terms analysis. In vivo, circulating FIAF protein was increased by the strain but this phenomenon was not correlated with modulation Fiaf expression in tissues (except a trend in distal small intestine).

Conclusion: We showed that Lactobacillus rhamnosus CNCMI-4317 induced Fiaf expression in human IECs, and increased circulating FIAF protein level in mice. Moreover, this effect was accompanied by transcriptome modulation of several pathways including immune response and metabolism in vitro.

No MeSH data available.


Related in: MedlinePlus