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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

L. rhamnosus CNCMI–4317 may induce Fiaf in a PPAR-α independent (a) but PPAR-γ dependent (b) manner. The antagonists (GW7647 and GW9662) were respectively added at 1 and 10μM 1h before challenging with agonists (GW6471 and rosiglitazone) during 6h.Bars represent means of Fiaf relative expression (percentage of rosiglitazone and GW7647 respectively) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene and by GW7647 (a) or rosiglitazone (b). Stars represent p<0.05 (*), p< 0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represent a non significant difference between L. rhamnosus CNCMI- 4317 versus L. rhamnosus CNCMI—4317 supplemented with GW9662.
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pone.0138880.g002: L. rhamnosus CNCMI–4317 may induce Fiaf in a PPAR-α independent (a) but PPAR-γ dependent (b) manner. The antagonists (GW7647 and GW9662) were respectively added at 1 and 10μM 1h before challenging with agonists (GW6471 and rosiglitazone) during 6h.Bars represent means of Fiaf relative expression (percentage of rosiglitazone and GW7647 respectively) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene and by GW7647 (a) or rosiglitazone (b). Stars represent p<0.05 (*), p< 0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represent a non significant difference between L. rhamnosus CNCMI- 4317 versus L. rhamnosus CNCMI—4317 supplemented with GW9662.

Mentions: Since Fiaf gene expression is controlled by both PPAR-γ and PPAR-α, we used specific ligands and inhibitors to investigate how L. rhamnosus CNCMI–4317 strain regulated Fiaf expression. Rosiglitazone and GW7647, agonists of PPAR-γ and PPAR-α respectively, increased the expression of Fiaf (P<0.001; Fig 2), whereas GW9662 and GW6471, the antagonists of PPAR-γ and PPAR-α respectively, strongly inhibited Fiaf gene expression (P<0.001). Additionally, L. rhamnosus CNCMI–4317 significantly induced Fiaf expression, which was completely abolished in the presence of GW6471 (P<0.05), indicating a PPAR-α dependent activation (Fig 2a). On the contrary, GW9662 did not modify Fiaf expression induced by the bacterial strain, suggesting that its effect was PPAR-γ independent (Fig 2b).


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)

L. rhamnosus CNCMI–4317 may induce Fiaf in a PPAR-α independent (a) but PPAR-γ dependent (b) manner. The antagonists (GW7647 and GW9662) were respectively added at 1 and 10μM 1h before challenging with agonists (GW6471 and rosiglitazone) during 6h.Bars represent means of Fiaf relative expression (percentage of rosiglitazone and GW7647 respectively) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene and by GW7647 (a) or rosiglitazone (b). Stars represent p<0.05 (*), p< 0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represent a non significant difference between L. rhamnosus CNCMI- 4317 versus L. rhamnosus CNCMI—4317 supplemented with GW9662.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0138880.g002: L. rhamnosus CNCMI–4317 may induce Fiaf in a PPAR-α independent (a) but PPAR-γ dependent (b) manner. The antagonists (GW7647 and GW9662) were respectively added at 1 and 10μM 1h before challenging with agonists (GW6471 and rosiglitazone) during 6h.Bars represent means of Fiaf relative expression (percentage of rosiglitazone and GW7647 respectively) from three independent experiments performed in triplicates. Data are normalized using β-Actin as control gene and by GW7647 (a) or rosiglitazone (b). Stars represent p<0.05 (*), p< 0.01 (**) and p<0.001 (***) in comparison with negative control (DMEM). ns represent a non significant difference between L. rhamnosus CNCMI- 4317 versus L. rhamnosus CNCMI—4317 supplemented with GW9662.
Mentions: Since Fiaf gene expression is controlled by both PPAR-γ and PPAR-α, we used specific ligands and inhibitors to investigate how L. rhamnosus CNCMI–4317 strain regulated Fiaf expression. Rosiglitazone and GW7647, agonists of PPAR-γ and PPAR-α respectively, increased the expression of Fiaf (P<0.001; Fig 2), whereas GW9662 and GW6471, the antagonists of PPAR-γ and PPAR-α respectively, strongly inhibited Fiaf gene expression (P<0.001). Additionally, L. rhamnosus CNCMI–4317 significantly induced Fiaf expression, which was completely abolished in the presence of GW6471 (P<0.05), indicating a PPAR-α dependent activation (Fig 2a). On the contrary, GW9662 did not modify Fiaf expression induced by the bacterial strain, suggesting that its effect was PPAR-γ independent (Fig 2b).

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