Volume 14, Numéro 3-4, Mai-Août 2007
|148 - 154
|Metabolism of PUFAs
|Publié en ligne
|15 mai 2007
Conversion of n-3 polyunsaturated fatty acids (PUFAs) and incorporation of docosahexaenoic acid (DHA) in cultured neural cells
Unité de Nutrition et Régulation Lipidique des Fonctions Cérébrales (Nu.Ré.Li.Ce), Institut National de la Recherche Agronomique (INRA), CRJ, 78352 Jouy-en-Josas cedex
Docosahexaenoic acid (DHA, 22:6n-3) in membrane phospholipids originates from dietary intake of preformed DHA and from conversion of its essential precursor α-linolenic acid (ALA, 18:3n-3). Cultured cells, especially nervous cells, are increasingly used to explore the uptake, metabolism and gene transcription effects of n-3 fatty acids, raising the question of the specific metabolic fate of different fatty acids and of the physiological relevance of their concentrations in the culture medium. This paper reports experimental data that 1) compare the dose-dependent incorporation of preformed DHA into the ethanolamine phosphoglycerolipids (EPG) of neural and cerebral endothelial cells in culture with that of the developing rat brain, 2) evaluate the pathway of DHA synthesis from ALA, eicosapentaenoic acid (EPA, 20:5n-3) or n-3 docosapentaenoic acid (DPA, 22:5n-3) in a model of neuronal cells, the SH-5YSY human neuroblastoma cells, and 3) characterize in these cells the mRNA expression profile of genes involved in the fatty acid metabolism. The incorporation of preformed DHA in EPG followed, both in vivo and in vitro, a dose-response curve from which two parameters were drawn: the DHAmax, i.e. the plateau-value of the linearized dose-response curve (expressed in weight % of total fatty acids), and the DHA50, the concentration of DHA in the diet or in the culture medium corresponding to an incorporation of DHA in EPG equal to one-half the DHAmax. The ratio of DHAmax to DHA50 reflects the propensity (so-called the ‘avidity’ for DHA) of cells or tissues to incorporate the exogenous DHA. The DHAmax and the DHAmax/DHA50 ratio values of SH-SY5Y cells and of rat brain endothelial cells in culture were compared to those of the frontal cortex and hippocampus of rats chronically deficient in n-3 fatty acids and supplemented with preformed DHA. The same DHAmax/DHA50 ratio values were found in SH-SY5Y (5.2) cells and in rat brain areas (5.1-5.7) when the DHA doses were expressed in lmol DHA/liter of culture medium and in lmol DHA/10 g diet, respectively. The SH-SY5Y cells were able to produce neoformed EPA, DPA and DHA from supplemental ALA. The incorporation of neoformed EPA and DPA in EPG followed a dose-response saturating curve, while that of DHA was bell-shaped. The different pattern of neoformed DPA and neoformed DHA suggests that the conversion pathway was limited at the terminal step of DHA synthesis. The mRNA profile showed that two enzymes of the peroxisomal b-oxidation system, the L- and D-bifunctional proteins, were expressed at lower levels than those of the endoplasmic reticulum pathway (Δ6-desaturase). These data show that incorporation of preformed DHA in cultured neuroblastoma cells match physiological values, indicating that DHA uptake, acyl-CoA activation, and phospholipid acyltransferases are active. However, the synthesis and incorporation of newly formed DHA in SH-SY5Y cells responds to a critical concentration-window of precursors which could originate from the low basal expression level of peroxisomal enzymes.
Key words: docosahexaenoic acid (DHA) / endothelial cells / ethanolamine phosphoglycerolipids (EPG) / n-3 PUFA conversion / peroxisomal enzymes / rat brain / SH-SY5Y neuroblastoma cells
© John Libbey Eurotext 2007
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