Comparison of docosahexaenoic acid uptake in murine cardiomyocyte culture and tissue: significance to physiologically relevant studies.

Long-chain n-3 (or omega 3) fatty acids, namely docosahexaenoic acid (DHA, 22:6n-3) and eicosapentaenoic acid (EPA, 20:5n-3) have been attributed cardioprotective properties. In this study, we evaluated the incorporation of DHA into cardiomyocytes and the shift in the omega 3/omega 6 ratio after supplementation of primary cardiomyocyte culture. Results are compared with atrial tissue concentrations attained after prolonged feeding of rats. The major difference between in vitro vs. in vivo supplementation is the paradoxical accumulation of arachidonic acid in cultured cardiomyocyte. However, this increase does not give rise to a higher PGE2 production after cellular stimulation, as compared with controls, possibly because of the associated inhibition of sPLA2 by DHA. Notably, in vitro supplementations with DHA 10 to 25µM approximate in vivo pharmacological treatments.

Differential distribution of DHA-phospholipids in rat brain after feeding: A lipidomic approach.

On a per-weight basis, the brain is the organ richest in lipids, including a remarkable proportion of polyunsaturated fatty acids (PUFAs) of the omega 3 series, namely eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. The cerebral effects of exogenous DHA likely depend on its degree of incorporation into neuronal phospholipids and on its distribution among the various brain structures, after intake. Hence, because PUFAs are not evenly distributed among the brain phospholipid classes and because the existence of class-specific phospholipases that regulate their turnover, we sought to investigate the incorporation of omega 3 PUFAs in selected brain areas regions and specific phospholipid classes. Rats (n=7) were administered, by oral gavage, 100mg/kg/d of a commercially available fish oil (containing ∼84% of long-chain omega 3 fatty acids, of which ∼38% of DHA and ∼46% of EPA). Control rats (n=7) received liquid paraffin. This treatment was continued for 30 days. Thereafter, we dissected three areas, namely the hippocampus, the striatum, and the cortex. Quantization of individual phospholipid classes and their molecular species was performed by ESI-MS/MS. Principal component analysis was used to examine the variation of the molecular lipid profiles (as percentage) induced by omega 3 supplementation. Our results show that provision of omega 3 fatty acids to rats results in their incorporation into brain phospholipids, the extent of which is lower in the striatum as compared with cortex and hippocampus. These data might in part explain the mixed therapeutic results obtained in neurological disorders, many of which are likely region-specific.

Docosahexaenoic acid down-regulates endothelial Nox 4 through a sPLA2 signalling pathway.

We investigated the anti-inflammatory and antioxidant activities of docosahexaenoic acid (DHA) by evaluating its modulation of the two enzymes most involved in vascular inflammation, i.e. endothelial secreted phospholipase A(2) (sPLA(2)) and NADPH oxidase 4 (Nox) 4. Exposure of human aortic endothelial cells (HAECs) to DHA led to its preferential incorporation into outer leaflet phospholipids. Pre-treatment with DHA abolished HAECs stimulation induced by A23187 and Ang II, whereas the effects on IL-1beta treatment were less pronounced. Group V sPLA(2) RNA was similarly modulated by DHA supplementation. In addition, DHA decreased Nox 4 expression and activity; this effect was associated with reduced production of reactive oxygen species. Further, the use of specific inhibitors allowed demonstrating that group V sPLA(2) is involved in the down-regulation of Nox 4 expression and activity by DHA. This interplay is mediated by ERK and PKC.

Polyunsaturated fatty acids and cardiovascular disease.

Replacing saturated with polyunsaturated (PUFAs) rather than monounsaturated fatty acids or carbohydrates results in cardiovascular prevention over a wide range of intakes. The mechanisms by which PUFAs reduce cardiovascular risk are manifold, and the extent and precise nature of their activities is the subject of several investigations, spanning from in vitro mechanistic studies to human intervention trials. This article reviews the most up-to-date evidence of the association between PUFA consumption and reduced cardiovascular mortality.

Weight and plasma lipid control by decaffeinated green tea.

We investigated whether regular decaffeinated green tea intake could modulate body weight in an experimental model of obesity. Male leptin-deficient (ob/ob) mice and their C57BL/6J lean littermates (4 weeks of age; n 20/genotype) were assigned randomly to receive either decaffeinated green tea or vehicle, for 6 weeks. Body weights were recorded weekly and fluid intake was measured at each replacement. Blood was collected from the heart into collection tubes, with Li(+)-heparin as the anticoagulant. Administration of decaffeinated green tea to ob/ob mice significantly slowed their rate of weight gain, as compared with animals that were fed buffer alone. This effect is apparent after only 1 week of supplementation. No significant difference was recorded between C57BL/6J lean mice administrated decaffeinated green tea and those given buffer alone. Decaffeinated green tea consumption by ob/ob mice was also associated with significantly lower cholesterolemia, triglyceridemia, and adiponectin concentration. Fecal lipids did not change significantly throughout the experiment. In conclusion, administration of decaffeinated green tea might contribute to weight control and provides an opportunity for through-the-day consumption, without the excitatory effects of caffeine.

Polyunsaturated fatty acids as antioxidants.

The susceptibility of fatty acids to oxidation is thought to be directly dependent on their degree of unsaturation. However, some in vitro and in vivo studies suggest that the relation between chemical structure and susceptibility to oxidation is not as straightforward as hypothesized from theoretical viewpoints. Indeed, long chain polyunsaturated fatty acids (LC-PUFAs) might be less oxidizable than others under specific experimental conditions. We investigated the free radical-scavenging potential of PUFA and the production of reactive oxygen/nitrogen (ROS/RNS) species by human aortic endothelial cells (HAECs) supplemented with different fatty acids. Fatty acid micelles scavenged superoxide in an unsaturation-dependent manner, up to eicosapentaenoic acid, which was the most effective fatty acid. Supplementation of HAEC with polyunsaturated fatty acids of the omega 3 series resulted in lower formation of ROS, as compared with cells supplemented with saturates, monounsaturates, or polyunsaturates of the omega 6 series. This effect was maximal at concentrations of 10muM. The effects of omega 3 fatty acids on reactive species production appear to be stronger when ROS were evaluated, as a milder, albeit significant effect was observed on RNS generation. Based on in vivo data showing reduced excretion of lipid peroxidation products after omega 3 intake and our data on ROS production and direct superoxide scavenging by LC-PUFAs, notably those of the omega 3 series, we propose that this series of fatty acid might act as indirect anti- rather than pro-oxidant in vascular endothelial cells, hence diminishing inflammation and, in turn, the risk of atherosclerosis and cardiovascular disease.