Combination of catheterized minipigs and high
throughput « omics » methodologies : For new paradigms
in the kinetics of development of insulin-resistance
1. Combination of urinary metabolome and new targeted plama metabolites (other than glucose and insulin)
: highlight key early time points involved in metabolic adaptations to the high fat – high sucrose diet
2. Some specific metabolites (urinary and other plasma metabolites) have been identified and described to
explain these early adaptations
3. These preleminary results will be the basis for experiments aiming at investigating the mechanisms
explaining the metabolic flexibility/inflexibility involved in the early phases of insulin resistance
development
S Polakof, D Rémond, M Rambeau, E Pujos-Guillot, JL Sébédio, D Dardevet, B Comte, I Savary-Auzeloux
UMR 1019 Unité de Nutrition Humaine, UNH, CRNH Auvergne, F-63000 CLERMONT-FERRAND
UMR 1019 Unité Nutrition Humaine UMR 1019 Unité Nutrition Humaine UMR 1019 UMR 1019 Unité Nutrition Humaine Unité Nutrition Humaine D0 D7 D14 D30 D60
Urinary metabolomics analysis (UPLC-QToF)
Plasma biochemical analysis Fasting blood sampling
Urine collection over the postprandial period
(0-8h after the meal)
6 adult Yucatan
minipigs
High fat / high sucrose diet (HF-HS)
Diet : ME/Kg : 15.6 MJ/Kg feed
Daily ingested ME : 13.3 MJ
Control diet (C)
Diet : EM/kg : 13.3 MJ/kg feed
Daily ingested ME : 5.3 MJ
D0
D60
1) Investigation of the metabolic shift (insulin resistance
installation) usually measured in plasma in the fasting state
(glucose and insulin)
2) Idea : Analysis of the subtle and early changes of the metabolic
trajectories using an integrative approach : metabolomics in a
compartiment integrating fasting and fed states : urine
Aim :
Study of the nutritional transition leading to the
installation of insulin resistance : EARLY PHASES
10 15 20 25 30 35 40 0 10 20 30 40 50 60 W eigh t (kg) Days Pigs' weight
+ 71%
0 1 2 3 4 5 6 7 8 0 7 14 mMFasting plasma glucose
a a b 0 0,1 0,2 0,3 0,4 0,5 0 7 days 15 days ng /mL
Fasting plasma Insulin
a a
b
Detection of insulin resistance based on classical biochemical markers at 15 days following HF-HS diet.
After 7 days, these biochemical markers remained unchanged and allow to conclude for a normal glucose
homeostasis and insulin sensitivity.
Time (days) Time (days)
1/ Detection of insulin resistance with the « classical plasma
biomarkers » : fasting glucose and insulin
Multivariate model OPLS-DA
Q2 0.3 R2 0.6 1382 significant ions
(total 5488)
Multivariate model OPLS-DA
Q2 0.7 R2 0.99 Initial homeostatic state (healthy) Transient homeostatic state (adaptation)
New homeostatic state (beginning of insulin resistance) Analysis focussed on D0, D7 and D14 Analysis including D0, D7, D14, D30, D60
2/ Metabolic trajectories following HF-HS diet: Analysis of targeted metabolites in urine
0 100 200 300 0 20 40 60 Intens ity
Urine citric acid
Time (days)
Molecules related to the transient / short term adaptations (D0-D7)
0 500 1000 1500 0 20 40 60 Intens ity Time (days)
Urine cholic acid
0 200 400 0 20 40 60 Intens ity Time (days) Urine Tyrosine 0 2000 4000 0 20 40 60 Intens ity Time (days)
Urine Hippuric acid
VIP=1.2 P=0.003 VIP=1.6 P=0.04 VIP=1.3 p=0.013 0 200 400 600 0 20 40 60 Intens ity Time (days) Urine Tryptophan VIP=1 p=0.006 VIP=1.4 p=0.06 Molecules presenting an
intermediary pattern of change
0 1 2 3 4 5 6 0 20 40 60 mM Time (days)
Arterial fasting urea
a
b ab
ab
Molecules related to the new homeostatic state (IR related
state : D7-D14) + plasma fasting glucose and insulin VIP=1.3 P=0.013 0 100 200 300 400 0 20 40 60 Intens ity Time (days)
Urine Pantothenic acid
0 0,4 0,8 1,2 0 10 20 30 40 50 60 mM Time (Days)
Arterial fasting lactate
a ab
