n-3 long-chain fatty acids and regulation of glucose transport in two models of rat brain endothelial cells.
Several in vivo studies suggest that docosahexaenoic acid (22:6 n-3), the main n-3 long-chain polyunsaturated fatty acids (LC-PUFA) of brain membranes, could be an important regulator of brain energy metabolism by affecting glucose utilization and the density of the two isoforms of the glucose transporter-1 (GLUT1) (endothelial and astrocytic). This study was conducted to test the hypothesis that 22:6 n-3 in membranes may modulate glucose metabolism in brain endothelial cells. It compared the impact of 22:6 n-3 and the other two main LC-PUFA, arachidonic acid (20:4 n-6) and eicosapentaenoic acid (20:5 n-3), on fatty acid composition of membrane phospholipids, glucose uptake and expression of 55-kDa GLUT1 isoform in two models of rat brain endothelial cells (RBEC), in primary culture and in the immortalized rat brain endothelial cell line RBE4. Without PUFA supplementation, both types of cerebral endothelial cells were depleted in 22:6 n-3, RBE4 being also particularly low in 20:4 n-6. After exposure to supplemental 20:4 n-6, 20:5 n-3 or 22:6 n-3 (15microM, i.e. a physiological dose), RBEC and RBE4 avidly incorporated these PUFA into their membrane phospholipids thereby resembling physiological conditions, i.e. the PUFA content of rat cerebral microvessels. However, RBE4 were unable to incorporate physiological level of 20:4 n-6. Basal glucose transport in RBEC (rate of [(3)H]-3-o-methylglucose uptake) was increased after 20:5 n-3 or 22:6 n-3 supplementation by 50% and 35%, respectively, whereas it was unchanged with 20:4 n-6. This increase of glucose transport was associated with an increased GLUT1 protein, while GLUT1 mRNA was not affected. The different PUFA did not impact on glucose uptake in RBE4. Due to alterations in n-6 PUFA metabolism and weak expression of GLUT1, RBE4 seems to be less adequate than RBEC to study PUFA metabolism and glucose transport in brain endothelial cells. Physiological doses of n-3 LC-PUFA have a direct and positive effect on glucose transport and GLUT1 density in RBEC that could partly explain decreased brain glucose utilization in n-3 PUFA-deprived rats.
INRA, UR909, Nutrition et Régulation Lipidique des fonctions Cérébrales, F-78352 Jouy-en Josas, France. pifferi@mnhn.fr <pifferi@mnhn.fr>
Fabien
F
Pifferi
Mélanie
M
Jouin
Jean-Marc
JM
Alessandri
Françoise
F
Roux
Nicolas
N
Perrière
Bénédicte
B
Langelier
Monique
M
Lavialle
Stephen
S
Cunnane
Philippe
P
Guesnet
D019325:3-O-Methylglucose / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry / Q000187:drug effects*; D002196:Capillaries / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005227:Fatty Acids / Q000032:analysis / Q000378:metabolism; D015525:Fatty Acids, Omega-3 / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism*; D051272:Glucose Transporter Type 1 / Q000096:biosynthesis / Q000235:genetics; D051274:Glucose Transporter Type 3 / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar
D017931:DNA Primers; D018076:DNA, Complementary; D005227:Fatty Acids; D015525:Fatty Acids, Omega-3; D051272:Glucose Transporter Type 1; D051274:Glucose Transporter Type 3; C495679:Slc2a1 protein, rat; C495690:Slc2a3 protein, rat; D019325:3-O-Methylglucose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005947:Glucose