Berry fruits and diabetes prevention Current trends and perspectives

Berry fruits and diabetes prevention Current trends and perspectives

Modulation of gut microbiota by plant polyphenols:
A Paradigm Shift in Understanding their Effects on
Diseases
Québec City, 22 October 2014
Yves Desjardins
@prof_yves
Institute of Nutrition and Functional Food
Laval University
Québec City, Québec, Canada
Why is bear poop blue ???
Health effects have been attributed to phenolic compounds
Blueberry Polyphenols
Anthocyanins
Quercetin
Procyanidins
Caffeic acid
Chlorogenic acid
P-coumaric adic
76
0%
0%
3% 2%
41%
36%
796
905
mg/100 g FW
18%
400
Rodrigues-Mateos et al. (2013) Amer. J. Clin Nutr. 98: 1179 .
⬇ CVD
⬇ LDL
⬆ Endothelial funct.
Reduction of inflammation
in many tissues and organs
Interaction with cell
cycle and induction
of apoptosis
Modulation of cellular
signaling cascades
(Cancer, CVD,
Diabetes, NDD)
Stimulation of
endogenous
antioxidant network
(SOD, Catalases)
Stylbenes
PAC
Ellagic
Acid
Antocyanins
Resveratrol
OPC
Ellagitannins
Induction of Phase 1
and 2 enzymes
⬇ cognitive decline
⬇ Metabolic syndrome
biomarkers
⬇ Insulin resistance
⬇ Glucose tolerance glucose
Bioavailability of polyphenols is very low
100 mg quercetin
0.3 - 0.7 µmol/l
150 mg catechin
0.1 µmol/l
200 mg hesperin
1.1 µmol/l
200 mg naringenin
200 mg anthocyanin
25 mg secoislariceresinol
6 µmol/l
~ 10 nmol/l
~ 30 nmol/l
Residence time in the body is relatively short
Polyphenols are recognized by the body as
xenobiotics and are rapidly eliminated
Brush border cells
Inhibition of
inflammation
2013
2008
2007
Modulation
of redox
signals
Inhibition of
endothelial
NADPH
oxidase
1999
Modulation of GUT
Microbiota
Preservation
of
Paraoxoanase
activity
Indirect
antioxidants
1995
Inhibition of
Xanthine ox.
1988
Antioxidants
1984
Modulation
of lipid
metabolism
Inhibition
NADPH
oxidase in
neutrophils
Inhibition
de
Nf-kB
Inhibition of
PPAR-g
Epigenetic
Modulators
Inhibition of
PARP
Anti-inflammatory
effects
Modulation
of neutrophil
function
Inhibition of
adhesion
molecules
expression
Anti-atherosclerotic
Compounds
Interference with
Arachidonic acid
metabolism
Modulation
of platelet
function
Direct antioxidant
activity
1950
In vivo
Decrease hypertension
In vitro
1944
1940
1936
Protection against
Capillary fragility
Isolation of vitamin P
Adapted de Weseler et al., 2012. JAFC
Gut microbes as factors involved in obesity
Our genome: the microbiome
Adapted from Backehed et al. Science 307, 1915-1920.
Metagenomic analysis of the colon microbiome
• 10X more bacterial cells than our body
• 100 more genes than our own genome.
• 1.5-2.0 kg microorganisms
• 100 trillion bacteria in our gut…
Microbial degradation of polyphenols
Flavonols
Flavanones
Hydroxyphenyl acetic acid
Hydroxyphenyl propionic acid
Flavanols
Phenyl valerolactone
Catechins
Hyppuric accid, Catecuic acid
Lignans
Isoflavones
Enterodiol
Equol
Procyanidins
p-coumaric acid ??
Cyanidin
Peonidin
Gonthier et al. 2003, Free Radical Biology & Medicine
Link between the microbiome and diabetes
(Esteves et al. 2011 Curr. Op. Clin. Nutr.)
Polyphenols
Prebiotics
Probiotics
 Bifidobacterium
Lactobacillus
Gut microbiome
Reduction in body
weight
Improvement in
insulin resistance
Increase tolerance
to glucose
Reduction of
inflammatory
biomarkers
Evolution of inflammation associated chronic deseases
Lean & Healthy
Obese & Diabetic
MCV
Sensitivity to insulin
Inflammation
Leptin
IL-1β
Adiponectin
MCP-1
MCP-1
+
+
+
MCP-1
ER Stress?
Necrosis
apoptosis?
iNOS
+
Chemokines Cytokines Adipokines
-
NO
Leptin
IL-6
Leptin
Adiponectin
APP’S
TNFα
IFN-γ
Lipopolysaccharide is the most
inflammatory molecule
recognized by the body
Polyphenols ?
Effect of polyphenols on gut microbes
Effect of polyphenols on gut microbes
Food Research International, 2013. DOI: 10.1016/j.foodres.2013.01.034
Ascending
Colon
Transversum
Colon
Decending
Colon
The gut microbiota plays an essential role in
the low-grade inflammation
Bacteroidetes
Firmicutes
Actinobacteria
Proteobacteria
Yves Desjardins
André Marette
Denis Roy
Emile Levy
Stéphanie Dudonné
Geneviève Pilon
Sébastien Matamoros
Gastrophénol Projet
Experimental design
MiceC57Bl6 N=12 per group
8 weeks of chow diet or HFHS
Chow
Gavage: vehicle
Drink: water
HFHS
Gavage: vehicle
Drink: water
HFHS
Gavage:
300 mg Cranberry
Drink:Water
Weight gain
Food intake
Gtt / Itt
Modulation of
microbiota
Bioavailability
Inflammation
Effect of cranberry extract on Weight Gain
Weight Gain
32.5
Body Weight (g)
30.0
Chow
HFHS
Cran
27.5
25.0
22.5
20.0
17.5
0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57
Time (Days)
Effect of cranberry extract on Total Energy Intake
Effect of cranberry extract on viceral adiposity
Effect of cranberry extract on glucose and insulin
tolerance
*
*
#
Effect of cranberry extract on glucose and insulin
tolerance during OGTT
*
#
*
#
Effect of cranberry extract on liver weight, triglycerides, lipid
peroxydation, plasma triglycerides and cholesterol
0.4
0.2
2
60
30
0
#
Plasma
Total Cholesterol
*
#
2
1
0
ho
C
ho
H w
FH
S
C
E
0.0
3
H w
FH
S
C
E
*
0.2
C
90
0
Total Cholesterol
(mM)
Plasma TG (mM)
0.4
4
C
C
Plasma
Triglycerides
0.6
6
ho
ho
H w
FH
S
C
E
0.0
MDA
0.6
8
120
H w
FH
S
C
E
#
150
pmol/mg proteins
TG
(mol/g of tissue)
Weight (g)
1.0
0.8
MDA
Liver Triglycerides
10
*
#
Liver Weight
#
Effect of a cranberry extract on hepatic steatosis
Chow
HFHS
+
Cranberry
HFHS
Effect of cranberry extract on Intestinal inflammatory
reaction
454 Pyrosequencing of the gut metagenome
• Analysis of the V6-V8 region of the
16S rRNA bacterial genes by highthroughput pyrosequencing
• (~ 100,000 reads on 1/8 plate)
• 20 samples sequenced
• (Mice DNA was pooled for each
diet and each week)
• A bar-code per sample allowed to
assign the obtained sequences
• 2566 sequences were obtained per
sample (after clean-up)
Evolution of the gut microbiota under a HFHS diet or
a diet supplemented with cranberry extract.
CHOW
HFHS
HFHS + cranberry
• True symbiont of
humans
• Represent 1-4% of intestinal
bacterial population
•
•
•
Mucus degrading bacteria
Produces SCFA – immunological
signals
Linked to obesity and low-grade
inflammation
Effect of cranberry extract on metabolic endotoxemia induced by
the HFHS diet
Plasma LPS
Plasma LPS (EU/mL)
0.6
*
*
0.4
*
*
**
0.2
Pr
H
FH
S
+
C
H
FH
S
+
S
FH
H
+
C
ra
ra
nb
nb
er
er
ry
ry
+
Pr
ob
io
tic
s
ob
io
tic
s
S
FH
H
C
H
O
W
0.0
N= 6 for the Chow, HFHS et HFHS + Cranberry
*p < 0.05 vs. Chow; **p < 0.05 vs. HFHS
2.0
#
1.5
1.0
0.5
0.0
1.5
1.0
0.5
0.0
0.0
Muc2
2.5
2.0
#
1.5
1.0
0.5
0.0
Ch
HFow
HS
CE
2.5
2.5
Ch
HFow
HS
CE
0.5
2.0
Relative expression
Muc2
Relative expression
1.0
Ch
HFow
HS
CE
1.5
Relative expression
2.0
Relative expression
Ch
HFow
HS
CE
2.5
Ch
HFow
HS
CE
Ch
HFow
HS
CE
COLON
Relative expression
JEJUNUM
Relative expression
Figure S3
Klf4
Reg3g
2.5
2.0
1.5
1.0
0.5
0.0
Klf4
Reg3g
2.5
2.0
1.5
1.0
0.5
0.0
PAC
PAC
Akkermansia ???
Mucus Production ?
T-Lymphoid Cells
PAC
PAC
Why is bear poop blue ???
High nM/low µM
Polyphenols
Resveratrol/Quercetin/Ferulic acid
HEALTH
(5%)
(95%)
Anti-microbial
Quorum sensing
Ion-binding
Aggregation
Phase I & II
Metabolites
Microbial
Metabolites
Gut-Brain axis
mM
Chemical cues about the qualityof the diet
Physiological
Activities
?
?
⬆︎Genes activation
Equol/Valerolactones/Coumaric acid
Urolithins/HBA
Prebiotic effects
SCFA
Gut Microbiota
DC
Tcell
Modified ecology
Defensins
⬆︎Nutrient processing
⬆ Akkermansia
⬆︎Mucus production
⬆︎Tight-junctions
Cytokines
⬆︎Immunity
⬇ ︎low-grade
⬇ ⬇︎
LPS
low-grade
Inflammation
inflammation
Gut-liver axis
⬇︎Cholesterol
⬇︎TG
Acknowledgements
Y. Desjardins
A. Marette
D. Roy
Stéphanie Dudonné
Pascale Dubé
Véronique Richard
Geneviève Pilon
Philippe St-Pierre
Fernando Arhe
Bruno Marcotte
Sébastien Matamoros
Thibault Varin
E. Levy
M.-C. Denis
Carole Garofalo
Quentin Lemoyne