Effects of eicosapentaenoic acid and docosahexaenoic acid on plasma membrane fluidity of aortic endothelial cells

Abstract

We investigated the relative effects of n-3 eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) on the plasma membrane fluidity of endothelial cells (EC) cultured from the thoracic aorta by determining fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and its cationic derivative trimethylamino-DPH (TMA-DPH). Fluidity assessed by TMA-DPH demonstrated no significant differences in plasma membranes of vehicle (dimethyl sulfoxide; DMSO)-, EPA-, and DHA-treated EC. Plasma membrane fluidity assessed by DPH polarization, however, was significantly higher in the order of DHA > EPA > DMSO. Total cholesterol content decreased significantly by 28.4 and 15.9% in the plasma membranes of DHA- and EPA-treated cells, respectively. Total phospholipid content remained unaltered in the plasma membranes of the three groups of cells; however, the molar ratio of total cholesterol to phospholipid decreased significantly only in the membranes of DHA-treated EC. The unsaturation index in the plasma membranes of EPA- and DHA-treated cells increased by 35.7 and 64.3%, respectively, compared with that in the plasma membranes of control cells. The activities of catalase and glutathione peroxidase in the whole-cell homogenates, and levels of lipid peroxides in either the whole-cell homogenates or in plasma membrane fractions were not altered in EPA- or DHA-treated EC. These results indicate that the influence of DHA is greater than that of EPA in increasing plasma membrane fluidity of vascular EC. We speculate that the greater effect of DHA compared to EPA is due to its greater ability to decrease membrane cholesterol content or the cholesterol/phospholipid molar ratio, or both, and also to its greater ability in elevating the unsaturation index in the plasma membranes of EC.