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Designed by Júlia Muñoz Carreras

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Molecular mechanisms involved in the protective

effect of the Mediterranean diet and olive oil

consumption in humans

Valentini Konstantinidou, PhD

PhD in Biomedicine, Universitat Pompeu Fabra, Department of Experimental and Health Sciences,

Barcelona, Spain

Thesis director: Dr. Maria Isabel Covas Planells

Cardiovascular Risk and Nutrition Group, Institut Municipal d´Investigació Mèdica (IMIM-Hospital del Mar)

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Presentation Outline

Introduction in Nutrigenomics

Objective-Hypothesis Methods and Results

Conclusions

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Introduction – Nutrigenomic era

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Principles

Introduction – Nutrigenomics

Müller and Kersten, Nature Reviews 2003

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Nutrigenomic studies have focused on investigating the

molecular mechanisms of action of several foods and nutrients,

particularly lipids, on cardiovascular risk factors and other

complex traits.

Intervention studies, in which subjects receive a controlled

dietary intake, provide the best approach for conducting cause-

effect relationships between gene expression and diet.

Limitations:

- Small number of participants

- Brief intervention duration

- Lack of replicationOrdovas et al. Mol Nutr Food Res 2007

Introduction – Nutritional Genomics

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Introduction – Nutrigenomic studies general

Mechanisms by which dietary patterns, foods or food components elicit their beneficial/harmful effects on human health are partially unknown.

One of the mechanisms could be the modulation of disease-related genes by dietary patterns, foods or food components.

At present, the knowledge concerning the role of diet in modulating atherosclerosis-related genes is limited.

State of the art

To understand the molecular mechanisms of genes-diet interaction.

To prevent diet-related diseases.

To develop evidence-based nutrition.

To contribute to public health.

Goals

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Introduction – Nutrigenomic era

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In peripheral blood mononuclear cells (PBMNCs)

• Higher increase in TNF expression after butter-rich breakfast vs olive oil.

• Higher increase in IL6 mRNA response after butter-rich breakfast vs walnut. (Jimenez-Gomez et al. Atherosclerosis 2009)

• Postprandial activation of NF B after butter- and walnut-rich meals vs olive oil.(Bellido et al. Am J Clin Nutr 2004)

In human adipocytes

• Anti-inflammatory gene expression profile after MUFA-diet vs SFA-diet.(van Dijk et al. Am J Clin Nutr 2009)

• Higher TNF expression after conjugated linoleic acid (CLA) vs olive oil.

• Lower expression in GLUT4 (glucose transporter 4), LEPTIN, LPL (lipoprotein

lipase) after CLA vs olive oil.(Raff et al. J Nutr 2009)

Introduction – Nutrigenomic studies II

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The consumption of a TMD, VOO and its PC, can modify the human in vivo gene expression.

The gene expression changes will be

towards a protective mode for

cardiovascular disease development.

Hypothesis

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Methods - Task 1: VOO intervention study-pilot (n=11)

1-4 days: habitual diet controlling excess of antioxidants, sunflower oil for raw and cooking purposes

5-7 days: diet with very low phenolic content, sunflower oil for raw and cooking purposes

Wash out period

Samples collection

Intervention day

50ml of VOO ingestion

1h 6h0h

•

• 3 weeks (Khymenets et al. OMICS 2009)

Design : Linear Study

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Mononuclear Transcriptome Response after Sustained Virgin

Olive oil Consumption in humansKhymenets, O. et al. (2009)

OMICS 13, 7-19

Characterization of Human Gene Expression Changes after Olive Oil Ingestion: an

Exploratory Approach. Konstantinidou, V. et al. (2009) Folia Biologica (Praha) 55, 85-91

7 insulin sensitivity-related genes were modulated by VOO ingestion

Results - Task 1: VOO intervention study

Task 1.2 Postprandial time course of changes in the expression of those genes

after VOO ingestion (qRT-PCR)

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Methods Characterization of Human Gene

Expression Changes after Olive Oil Ingestion: an Exploratory Approach.

Konstantinidou et al. Folia Biologica (Praha) 2009

Time Course of Changes in the Expression of Insulin-Sensitivity Related Genes after an Acute

Load of Virgin Olive Oil.Konstantinidou, V. et al.

OMICS 2009

Selection of 47

atherosclerosis-related genes

Literature review

Mononuclear Transcriptome Response after Sustained Virgin Olive oil

Consumption in humans. Khymenets et al. OMICS 2009

Task 2: TMD intervention

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Methods - Task 2: TMD intervention study Study Flow diagram

Invited to be screened

(n=99)

Ineligible (n=9)

Did not meet inclusion criteria (n=7)Declined to participate (n=2)

Randomly assigned

(n=90)

Declined to follow up (n=1)

Control Group (n=29)

(n=20 in gene expression)

TMD+WOO (n=30)

(n=16 in gene expression)

TMD + VOO (n=30)

(n=20 in gene expression)

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Results - Task 2: TMD intervention study, Three-group analysesGene expression changes after TMD+VOO intervention

p<0.05 for linear trend in all cases * p<0.05 vs. control

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Tyr

osol

(n

g/m

l)

p=0.007 for quadratic trend

*

†

Control group TMD+WOO TMD+VOO

Hyd

roxy

tyro

sol (

ng/

ml)

†

Control group TMD+WOO TMD+VOO

Results - Task 2: TMD intervention study Volunteer’s dietary compliance

• Volunteers compliance was good as it is reflected in urinary T and HT levels

* p<0.05 vs control; † p<0.05 vs TMD+WOO

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Conclusions

Strengths

To work in real life conditions in all interventions

To use both the whole dietary pattern and the single component approach

To work against our hypothesis by using a Spanish dietary pattern in the control group

Limitations

- A lack of control group for the VOO intervention

- The inability to assess potential interactions

- As expected, we observed modest changes in gene expression

- Unknown effects over longer intervention periods

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Conslucions

POLK

LIAS

CD36

PPARBP

ADRB2

ADAM17

ALOX5AP

IFNγ

IL7R

ARHGAP15

OGT

Inflammation Oxidative stress Insulin resistance

The gene expression changes were observed in a protective

mode for counteracting these situations.

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Discussion

These results provide, for the first time, evidence on:

An in vivo human nutrigenomic effects of the TMD, in healthy

volunteers.

An in vivo human nutrigenomic effect of olive oil phenolic

compounds down-regulating atherosclerosis-related genes.

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Thank you for your attention