PLACENTAL TRANSFER OF FATTY ACIDS AND FETAL IMPLICATIONS

Dr. Elvira Larquéelvirada@um.es

Department of PhysiologyUnversity of Murcia (Spain)

Note: for non-commercial purposes only

FETAL PROGRAMMING

BRAIN60% FAT (dry matter)

~ 40% LONG CHAIN POLYUNSATURATED FATTY ACIDS (LC-PUFA):

- 15% DOCOSAHEΧAENOIC ACID (DHA,22:6 n-3 ): Neuronal development

- 10% ARACHIDONIC ACID (AA, 20:4 n-6 ) : Infantile Growth

Polyunsaturated FA Synthesis

22:6ωωωω-3docosahexaenoic

18:2ωωωω-6 18:3ωωωω-3αααα-linolenic

18:3ωωωω-6γγγγ-linolenic

20:4ωωωω-6arachidonic

20:5ωωωω-3eicosapentaenoic

∆∆∆∆6-desaturase

elongase & ∆∆∆∆5−−−−desaturase & elongase

PROSTA-GLANDINS

MEMBRANES

linoleic

EFA and LC-PUFA PROVIDED BY PLACENTAL TRANSFER

EFA:

LC-PUFA:

FFA

FFA

PasiveDiffusion

Facilitated

Transport

Maternal circulation

Trofoblast cytosol FFA FFA

membrane

FFA

Lipoprotein

R

?

PLA2 Lipases

AGL

Lipase

EL

PL

EL, Endothelial lipase

Fatty Acid Placental Uptake

FABPpmFATP, FAT

Lipase

LPL

TG TG

TG

LPL, Lipoprotein lipase

• Placental transfer of fatty acids is a complex processthat includes theirbinding to membraneand cytosolic proteincarriers

Fatty Acid PlacentalTransfer

p-FABPpm (placental plasma fatty acid binding protein) not sequenced??.

m a te rn a l co rd

S A T 4 7 .1 (2 .5 ) 4 9 .9 (1 .7 )

M U F A 1 4 .9 (2 .7 ) 1 3 .4 (2 .2 )

1 8 :2 n -6 2 1 .0 (3 .4 ) 7 .4 (1 .4 )

1 8 :3 n -3 0 .2 0 (0 .1 0 ) 0 .0 0 (0 .0 3 )

2 0 :4 n -6 7 .7 (1 .9 ) 1 6 .1 (2 .5 )

2 2 :6 n -3 2 .9 (1 .0 ) 4 .8 (1 .7 )

Fatty- Acid Percentages of Maternal and Cord Plasma Phosph olipids [median (IQR)]

Koletzko et al. Eur J Pediatr 1998

BIOMAGNIFICATION OF LC-PUFA

(LA)

(ALA)

(AA)

(DHA)

PERILIP PROJECT (FP6 EU)

Material y methods

Single oral dose of stable isotope tracer in a sugar cube:- 13C-PA (Palmitic acid: saturated FA): 0.5 mg/Kg- 13C-OA (Oleic acid: monounsaturated FA): 0.5 mg/Kg- 13C-LA (Linoleic acid: essential FA): 0.5 mg/kg

- 13C-DHA (Docosahexaenoic acid, LC-PUFA): 0.1 mg/kg

� Study 4 hours before cesarean section (n=4)

� Study 12 hours before cesarean section (n=11)

In vivo evaluation of the placental transfer of fatty acid slabeled with stable isotopes in humans

Aim

Stable Isotope Study on FA Placental Transfer

4h FA Tracer Study

-4 -3 -2 -1 0 10,0

0,2

0,4

0,6

0,8

1,0

DHA

LA

OA

PA

13C

-AP

E (

%)

time (h)

-4 -3 -2 -1 0 10,0

0,2

0,4

0,6

0,8

1,0

13C

-AP

E (

%)

time (h)

-4 -3 -2 -1 0 10,00

0,01

0,02

0,03

0,04

13C

-AP

E (

%)

time (h)

Appearance of 13C-FA enrichment in maternal plasma Maternal Plasma TG Maternal Plasma FFA

Maternal Plasma PLNo enrichment in Maternal CholesterolEsters

N=4 (Larqué et al. J. Lipid Res., 2003)

Placenta/plasma fatty acid concentration in plasma (µmol/ml) * APE-plasma(%) Ratio fatty acid concentration in placenta (µmol/g) * APE-placenta

X 100

Distribution of tracer between placenta and plasma (M ± SEM)

13C-PA 13C-OA 13C-LA 13C-DHA0

5

10

15

20

25

30(%

) R

atio

pla

cent

a/pl

asm

a

bc

c

b

a

=

4h FA Tracer Study

(Larqué et al. J. Lipid Res., 2003)

Design of 12h FA tracer StudySubjects: 11 pregnant women, elective caesarian section

weight: 78 ± 8 kg height: 161± 6 cmage: 33 ± 4 years gestation: 39.8 ± 1 wk

Oral application of stable isotope tracer in a sugar cube:• palmitic acid (13C-PA): 0.5 mg/kg • oleic acid (13C-OA): 0.5 mg/kg• linoleic acid (13C-LA): 0.5 mg/kg• docosahexaenoic acid (13C-DHA): 0.1 mg/kg

- Maternal blood: 0h, 9h, 10,11h, 12h (c-section), 13h - Placenta collection and venous cord blood

13C-enrichment of FA by gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS)

Tracer administration (12h before caesarean section )

basal 9 10 11 12

0

2

4

6

8

10

12

14

(h)

basal 9 10 11 12

0

0,2

0,4

0,6

0,8

1

(h)

basal 9 10 11 12

0

2

4

6

8

10

12

14

16

(h)

basal 9 10 11 12

0

2

4

6

8

10

12

14

(h)

TGPL

FFACE

C-L

A(µ

mol

/l)13 C

-DH

A(µ

mol

/l)13

C-P

A(µ

mol

/l)13

C-O

A(µ

mol

/l)13

SATURATED FA MONOUNSATURATED FA

ESSENTIAL FA

a a

b bb

bcbc

aa

a

ab

bb

bb

LC-PUFA

P < 0.05

13C-FA CONCENTRATION IN MATERNAL PLASMA12

h F

A tr

acer

stud

y

0

2

4

6

8

10

12

14

16

PL

NEFA

TG

EC

13C-FA enrichment in placental TG and NEFA tended to be h igh

Phospholipids constituted about 80-90% of FA in placent a

13C-FA Concentration in Placenta

12h

a

a

a

ab

bb

bb b bcbbcc

C-F

A (

µm

ol/l)

13

13C-OA 13C-LA 13C-DHA13C-PA

Mean ± SD, P < 0.05

12h

FA

trac

erst

udy

12h FA tracer study

(%)

Rat

io p

lace

nta/

mat

erna

l pla

sma

0

50

100

150

200

250

300

350

400

450

500

4h

α=40,17 (DHA)

α=6,71( LA)α=4,75 (PA)α=4,18 (OA)

*

12h

* P<0,05

*

α = slope

(Gil-Sánchez, Larqué et al. Am. J. Clin. Nutr., 2010)

Distribution placenta/maternal plasma (M ± SEM)

0

10

20

30

40

50

(%)

Rat

io p

lace

nta/

mat

erna

l pla

sma

a

bb

b

Different letters: P<0,05

Placenta

13C-PA 13C-OA 13C-LA 13C-DHA

Comparison 4h vs 12h

Cord/plasma AUC FA concentration in plasma (µmol/ml) * APE-mat. plasma

(%) Ratio FA concentration in cord (µmol/g) * APE-cord plasmaX 100

Distribution Cord/Maternal Plasma

13C-PA 13C-OA 13C-LA 13C-DHA

(%)

Rat

io c

ord/

plas

ma

AU

C

a

bb b

Mean ± SD, P < 0.05

=

0

0,5

1

1,5

2

12h

12h

FA

trac

erst

udy

0

520

21

22

23

01234567

%-w

t/wt

Control 400µg MTHF 0.5mgDHA DHA+MTHF

PROYECTO NUHEAL: Placental PL

a a

b b

DHA AA

• Supplementation of 136 Spanish pregnant women from wk 20 to delivery

No significant changes in placental mRNA expression of FATP-1, FATP-4, FATP-6, FAT, FABPpm, H-FABP, B-FABP, A-FABP among the groups

NUHEAL: Correlations placental carriers-Placental PL

0

50

100

150

200

250

0 2 4 6 8 10

0102030405060708090

100

0 2 4 6 8 10

FA

TP

-1 (

AU

)

R = + 0.364; P = 0.001 R = + 0.387; P = 0.0007

FA

TP

-4 (

AU

)DHA (%) DHA (%)

DHA (%) IN MATERNAL PLASMA PL CORRELATES WITH PLACENTAL PROTEINS FATP-1 (r=0.32, p=0.001) AND FATP-4 (r=0.23, p=0.012)

(Larqué et al. Am J Clin Nutr. 2006)

FA

TP

-4 p

lace

nta

(AU

)

DHA(%) cord blood PL

r = + 0.20; P = 0.032*

(Larqué et al. Am J Clin Nutr. 2006)

0102030405060708090

100

0 5 10 15 20

NUHEAL: Correlations placental FA carriers-CordPlasma PL

Normal plasma LC-PUFA in women with gestational diabetes mellitus, but low erytrocyte PL LC-PUFA in their babies (Wijendran et al. 2000)

Pregestational or gestational diabetes mellitus was found toadversely affect attention span and motor functions of offspring at

school age (Ornoy et al. 2001)

GESTATIONAL DIABETES MELLITUS

Delayed brain maturity in these newborns compared to controls?

PREELIMINARY RESULTS

0102030405060708090

100

Maternal Cord(m

g/dl

)Control (n=22)

GestationalDiabetes (n=26)

*GLUCOSE

LC-PUFA n-3

*(%l)

0

2

4

6

8

10

12

Maternal Placenta Cord

**

DHA P =0.062

PREELIMINARY RESULTS

VENOUS CORD BLOOD

Control (n=22)

Gestational diabetes (n=26)

(%)

*

**

(%)

SATURATED (SFA)

EFA, LC-PUFA n-3

0

5

10

15

20

25

30

EFA LC-PUFA LC-PUFA n-3 DHA

0

5

10

15

20

25

30

35

40

45

SFA MUFA PUFA

PLACENTAL TRANSFER OF LABELED FA IN GESTATIONAL DIABETES ON GOING

NEURONAL DEVELOPMENT IN BABIES FROM GESTATIONAL

DIABETES MOTHERS DURING THE 1º YEAR OF LIFE

•Bayleys test (6 months and 12 months)

• Sleep/wake cycles by actigraphy methods

• Evaluation of circadian rhythms of temperatureusing temperature sensors.

Ondas medias de temperatura de la piel

32,00

32,50

33,00

33,50

34,00

34,50

35,00

35,50

36,00

9:00

10:2

0

11:4

0

13:0

0

14:2

0

15:4

0

17:0

0

18:2

0

19:4

0

21:0

0

22:2

0

23:4

0

1:00

2:20

3:40

5:00

6:20

7:40

Hora del día (hh:mm)

Tem

pera

tura

(ºC

)

15 días

30 días

90 días

Actimeter

EVOLUTION OF CHRONOBIOLOGICAL RHYTHMS

Temperaturesensor

32,0

32,5

33,0

33,5

34,0

34,5

35,0

35,5

15 days 30 days 3 months 12 years 20 years

Te

mp

era

ture

ºC

Age

Mesor

Different letters indicate significant differences by repeated measures ANOVA (for 15 days, 30 days and 3 months comparisons). Different numbers indicate significant differences when comparing babies (3 months) with child and adult groups.

EVOLUTION OF CORE BODY TEMPERATURE

ab a

b1

22

0,0

0,5

1,0

1,5

2,0

2,5

15 days 30 days 3 months 12 years 20 years

Te

mp

era

tu

re º

C

Age

M5-L10 Values

a

b1

1

2

ab

PREELIMINAR RESULTS

MEAN OF TEMPERATURE AMPLITUD OF TEMPERATURE

Sarabia et al. 2009. Acta Physiologica, 195 (667):101

Financial support by Alexander von Humboldt Foundation, HERO S.L. and by the Commission of the European Communities, within the FP 6 (PERILIP PROJECT and NUHEAL PROJECT).

This presentation does not necessarily reflect the views of the Commission and in no way anticipates its future policy in this area. Elvira Larqué is a recipient of the ESPGHAN Award for Young Investigators. Budapest. 2009.

Thank you for your kind attention!!!!

University of Murcia (SPAIN)Ana Pagán

María SabaterJ.A. Madrid

Salvador Zamora

UniversityUniversity ofof Murcia Murcia (SPAIN)(SPAIN)

Ana PagánMaría SabaterJ.A. Madrid

Salvador Zamora

Pediatric ServiceArrixaca Hospital

Murcia (Spain)Matilde ZornozaSilvia FuentesVicente Bosch

Manuel Sánchez-Solís

PediatricPediatric ServiceServiceArrixacaArrixaca HospitalHospital

Murcia (Murcia (SpainSpain))Matilde ZornozaSilvia FuentesVicente Bosch

Manuel Sánchez-Solís

Gynecology ServiceArrixaca Hospital

Murcia (SPAIN)Alfonso Gil-SánchezMaría Teresa PriteoJosé Eliseo BlancoJuan José Parrilla

GynecologyGynecology ServiceServiceArrixacaArrixaca Hospital Hospital

Murcia (SPAIN)Murcia (SPAIN)Alfonso Gil-SánchezMaría Teresa PriteoJosé Eliseo BlancoJuan José Parrilla

LMU Munich(GERMANY)Berthold Koletzko

Hans DemmelmairFabianne Hanebut

All staff

LMU MunichLMU Munich(GERMANY)(GERMANY)

Berthold KoletzkoHans DemmelmairFabianne Hanebut

All staff

THANK YOU !