Changes in plasma triacylglycerol concentrations after sequential lunch and dinner in healthy subjects.

OBJECTIVES: The present study examines the kinetic of plasma triacylglycerol (TAG) after sequential ingestion of lunch and dinner as well as the contribution of dietary fat ingested at lunch to subsequent post-dinner TAG composition. METHOD: Six healthy subjects were included. After standardized breakfast (7: 30AM), 2 mixed meals with fat loads composed of 44 g olive oil (rich in oleic acid) at lunch (12PM) and 44 g sunflower oil (rich in linoleic acid) at dinner (7PM) were ingested. [1-13C] palmitate was added in lunch only. Plasma TAG and chylomicron-TAG (CMTAG) levels were measured sequentially after meals. [1-13C] palmitate enrichment and concentrations of oleic acid and linoleic acid were measured in all lipid fractions. RESULT: Post-dinner plasma TAG peak was delayed as compared to lunch (3 hours vs 1 hour, p=0.002) whereas the magnitude of the postprandial peaks was not significantly different between lunch and dinner (2.4+/-0.3 vs 2.0+/-0.4 mmol/L, p=0.85). [1-13C] palmitate enrichment was maximal 5 hours after lunch in all lipid fractions and decreased slowly thereafter. After dinner ingestion, the rate of decline of [1-13C] palmitate enrichment plateaued during the first 60 minutes. Oleic acid increased slightly and immediately after dinner and remained the predominant fatty acid in all lipid fractions during the first hour after dinner. A delayed peak of plasma and CM-TAG was observed after dinner as compared to lunch without difference in the magnitude of peaks. CONCLUSION: The contribution of dietary fat ingested at lunch to post-dinner lipemia is confirmed despite the relatively long lasting interval between the 2 meals (7 h) and the absence of any early peak of plasma TAG after dinner.

Long-term supplementation of culture medium with essential fatty acids alters alpha-linolenic acid uptake in Caco-2 clone TC7.

We investigated the influence of four different culture media: 20% fetal bovine serum (FBS), 5% FBS, 5% FBS supplemented with 10 mg x L(-1) linoleic acid (18:2(n-6)) or alpha-linolenic acid (18:3(n-3)) on alpha-linolenic acid apical uptake in clone TC7 of human intestinal Caco-2 cell line. Neither cellular viability nor cell monolayer integrity and permeability were altered by the four culture conditions. Our results show that the different culture media led to changes in alpha-linolenic acid maximal rate of uptake (Vmax) but did not alter the apparent transport constant (Km). Reducing FBS concentration from 20% to 5% increased significantly the rate of alpha-linolenic acid uptake, which was further increased by supplementation of the medium with 18:2(n-6) or 18:3(n-3). Supplementation with essential fatty acids led to a marked enrichment of brush-border membrane phospholipids in polyunsaturated fatty acids of the corresponding series and decreased significantly the levels of monounsaturated fatty acids. Saturated fatty acids, unsaturation index, and cholesterol/fatty acid ratios were unchanged. No clear relation could be established between the changes in membrane lipid composition and the alterations of alpha-linolenic acid uptake. These results indicate a weak influence of membrane lipid composition in the modulation of the uptake. Therefore, the increase of uptake following long-term supplementation of TC7 cells with essential fatty acids could be attributed to an increase of the expression of membrane protein(s) involved in the apical uptake of long-chain fatty acids. This remains to be established.

Na(+)-H+ antiporter activity in relation to membrane fatty acid composition and cell proliferation.

Na(+)-H+ exchange activity was studied in a human breast cancer cell line. At steady state, the intracellular pH (pHi) of the cells was 7.23 +/- 0.01, and intracellular buffering capacity (beta i) was 44 +/- 4 mM/pH unit. pHi was controlled by a Na(+)-H+ antiporter that was reversible, electroneutral, inhibited by 5-(N-methyl-N-isobutyl)amiloride, and dependent on extracellular Na+. The exchanger function depended on internal H+ concentration, according to an allosteric activation mechanism obeying the model of Hill. The exchanger was inactive at pHi > or = 7.22, and its maximal activity was reached at pHi < 6.60. The exchanger was stimulated by osmotic shrinking but was unaffected by growth factors (epidermal growth factor, insulin-like growth factor I) or by serum. When cells were grown in a medium supplemented with linoleic or alpha-linolenic acids, large quantities of the additional fatty acid accumulated in membranes, saturated fatty acids increased, and monounsaturated fatty acids decreased. These changes reduced cell proliferation but had no effect on the steady-state value of pHi, on beta i, or on the kinetic parameters of the Na(+)-H+ exchanger. Therefore, in this system, cell proliferation is not directly related to the activation of the Na(+)-H+ exchanger.