• Aucun résultat trouvé

Impact of human milk pasteurization on digestive kinetics and intestinal lipid absorption using a combination of in vitro models

N/A
N/A
Protected

Academic year: 2021

Partager "Impact of human milk pasteurization on digestive kinetics and intestinal lipid absorption using a combination of in vitro models"

Copied!
2
0
0

Texte intégral

(1)

HAL Id: hal-02100438

https://hal.archives-ouvertes.fr/hal-02100438

Submitted on 2 Jun 2020

HAL is a multi-disciplinary open access

archive for the deposit and dissemination of

sci-entific research documents, whether they are

pub-lished or not. The documents may come from

teaching and research institutions in France or

abroad, or from public or private research centers.

L’archive ouverte pluridisciplinaire HAL, est

destinée au dépôt et à la diffusion de documents

scientifiques de niveau recherche, publiés ou non,

émanant des établissements d’enseignement et de

recherche français ou étrangers, des laboratoires

publics ou privés.

Impact of human milk pasteurization on digestive

kinetics and intestinal lipid absorption using a

combination of in vitro models

Olivia Ménard, Amélie Deglaire, Didier Dupont, J. Picaud, Armelle Penhoat,

Marie-Caroline Michalski

To cite this version:

Olivia Ménard, Amélie Deglaire, Didier Dupont, J. Picaud, Armelle Penhoat, et al.. Impact of human

milk pasteurization on digestive kinetics and intestinal lipid absorption using a combination of in

vitro models. 6.International Conference on Food Digestion, Apr 2019, Grenade, Spain. 2019.

�hal-02100438�

(2)

*

olivia.menard@inra.fr

Objective

Impact of human milk

pasteurization on premature infants’

digestion and intestinal lipid absorption

Maternal milk is the optimal food for neonates (WHO).

Hospitalized preterm newborns may be fed pasteurized donor milk if their own mother’s milk is unavailable. Holder pasteurization (62.5°C, 30 min) is conducted for sanitary reasons, but this alters immunological and bioactive components (Akinbi et al., 2010; Van Gysel et al., 2012). As a result, by denaturing the bile salt-stimulated lipase, fat absorption and growth may be reduced in preterm infants (Andersson et al., 2007; Thomas et al., 1999)

Olivia Menard

1*

, Marine Vincent

2,3

, Amélie Deglaire

1

,

Didier Dupont

1

, Jean-Charles Picaud

2,3

, Armelle Penhoat

2

, Marie-Caroline Michalski

2

1 STLO, INRA, Agrocampus Ouest. 35042 Rennes. France.

2Univ-Lyon, CarMeN Laboratory, INSERM U1060, INRA U1397, INSA Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne. France 3Department of Neonatology, Hôpital de la Croix-Rousse, Groupement Hospitalier Nord, Hospices Civils de Lyon, F-69004 Lyon. France

Infant static in vitro digestion model

Gastric phase (37°C)

Ratio Meal : Simulated Gastric Fluid of

63:37

Enzymes : Pepsin

(126 U/mL) and Lipase (10 U/mL)

from Rabbit Gastric Extract

60 min of digestion at pH 5.3

Intestinal phase (37°C)

Ratio Meal : Simulated Gastric Fluid of

39:61

Enzymes: porcine pancreatin (lipase

: 59 U/mL)

Bovine bile (1.6 mmol/L)

60 min of digestion at pH 6.6

Infant

No oral phase Adapted for

preterm infant of 2 kg

Ménard et al., 2018 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 0.01 0.1 1 10 100 1000 v o lu me (% ) size (µm) Raw milk G0 G60 I60

Particle size distribution

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 0.01 0.1 1 10 100 1000 v o lu me (% ) size (µm) Pasteurised milk G0 G60 I60

RHM

vs

PHM

lipolysis

• Before digestion: larger fat globules for pasteurized (D[4;3]=14.5µm) as compared to raw (D[4;3]=6.4µm ) is probably due to protein aggregation on globules during pasteurization. • At 60min of gastric digestion: particle aggregation in both conditions but less in PHM (D[4;3]=23µm) than in RHM (D[4;3]=48µm).

• At 60min of intestinal digestion, same size of mixed compounds (D[4;3]=13.4µm) in both conditions.

Holder pasteurization increases milk fat globule size probably due to protein aggregation around fat globules.

This may be responsible for the observed different kinetics of proteolysis, which could impact the release of some

bioactive peptides. Pasteurization did not impact triacylglycerol lipolysis but might unfavourably modulate

enterocytic lipid uptake, thus requiring further investigation.

Impact of human milk pasteurization on digestive kinetics and

intestinal absorption using a combination of in vitro models

RHM

vs

PHM

proteolysis

• No difference between RHM and PHM towards the disappearance of triacylglycerols (TAG) during gastro-intestinal digestion. • Lipolysis occurs mainly in the intestinal phase. 0 10 20 30 40 50 60 70 80 90 100 0 15 30 60 75 90 120 % o f re si d u a l i n te n sit y

digestion time (min) casein raw pasteurized intestinal gastric

*

*

*

TAG 2.5

µg0.25µg 5.0µg G0RMG60RMI15RM I30RMI60RM

G0 PM G60 PM I15 PM I30 PMI60PM FFA Cholesterol DAG MAG 1.0 µg 0.5 µg G0PM PL

Standards mix TAG+AGL

LIPOLYSIS

PROTEOLYSIS

STRUCTURE

0 10 20 30 40 50 60 70 80 90 100 0 15 30 60 75 90 120 % o f re si d u a l i n te n sit y

digestion time (min) lactoferrin raw pasteurized gastric intestinal

*

*

*

• Lactoferrin proteolysis is higher in PHM than in RHM at the end of gastro-intestinal digestion (residual lactoferrin of 26±4 % vs. 53±18%)

• β-casein is less susceptible to gastric proteolysis in PHM than in RHM (78±3 % vs. 37±4% remaining at 60 min of gastric digestion) but no difference in the intestinal phase with less than 5% of β-casein remaining.

• Same behaviour of α-lactalbumin for both types of milk with a resistance in the gastric phase and an extensive hydrolysis in the intestinal phase (less than 6% remaining at 60min of intestinal digestion). 0 10 20 30 40 50 60 70 80 90 100 0 15 30 60 75 90 120 % o f re si d u a l i n te n sit y

digestion time (min) α-lactalbumin raw pasteurized gastric intestinal

*

0 2 4 6 8 10 12 14 16 18 G0 G60 I60 SS (m² /g ) digestion time Specific surface evolution during digestion

raw pasteurized 0 10 20 30 40 50 60 70 80 90 100 0 60 75 90 120 % r e si dua l int e ns it y of T A G

dig time(min ) G+I

raw pasteurized

gastric intestinal

Sample characterization

Microscopic level : laser light scattering

Molecular level :

Thin Layer Chromatography (TLC)

Electrophoresis (SDS-Page)

Caco-2/TC7 cell line, derived from a human colon

adenocarcinoma, is a well-known lipid absorption

model.

Cells were incubated in Transwell® plates during 16

hours with diluted PHM vs RHM-digestion media.

Cell monolayer integrity was assessed by

transepithelial electrical resistance and occludine

immunostaining.

Intracellular lipid droplets could be visualized using

Oil Red O staining.

In vitro intestinal absorption model

LIPID ABSORPTION

• Incubation of digested human milk on Caco2/TC7 cells did not alter cell monolayer integrity.

• Less lipid droplets seemed to be present within cells incubated with PHM compared to RHM.

RHM

PHM

*: significant difference between RHM and PHM p<0.05 (t.test)

Raw

Human Milk

(RHM)

Pool of human milk

(n=2 donors)

Pasteurized

Human Milk

Références

Documents relatifs

Free fatty acids (FFA) released during the simulated duodenal digestion of raw oat groat (yellow lines), flakes (red lines) and flour (blue lines) at concentrations corresponding

The digestion of caseins and soluble proteins differed in both the intestinal kinetics and the molecular weight of the peptides in the jejunum.. The differential intestinal

[r]

Here we compare triglyceride age and, thereby, triglyceride turnover in vWAT and sWAT biopsies from 346 individuals and find that subcutaneous triglyceride age and storage capacity

P. Michel Laplaud, Dominique Bauchart, Denys Durand, M. Lipoproteins and apolipoproteins in intestinal lymph of the preruminant calf, Bos spp., at peak lipid absorption.. We

The aim is to apply analytical tools proper of viability theory and convex analysis, see [6], [8], [19], [20], to determine a computation-oriented criterion for characterizing

We construct a partitioning oracle that given query access to a graph from a specific family of graphs, provides query access to a fixed partition, and queries a

Nous présentons dans ce chapitre un modèle d’aide à la décision pour aider les maraîchers à sélectionner les clients les plus rentables compte-tenu de la production de la ferme,