Effect of aging on plasma membrane fluidity of rat aortic endothelial cells

To assess age-related changes in the physical properties of vascular endothelial cell (EC) plasma membranes, we measured membrane fluidity with 1,6-diphenyl-1,3,5-hexatriene (DPH), 1-(4-trimethylammonium-phenyl)-6-phenyl-1,3,5-hexatriene, and 10-(1-pyrene)dodecanoic acid, and investigated the parameters affecting membrane fluidity of endothelial cells (ECs) cultured from the thoracic aortas of young (5-week-old) and aged (100-week-old) rats. Plasma membrane fluidity of aged rat ECs was significantly lower than that of young rat ECs, as assessed by increased 1,6-diphenyl-1,3,5-hexatriene fluorescence polarization and by decreased pyrene excimer formation, although 1-(4-trimethylammonium-phenyl)-6-phenyl-1,3,5-hexatriene did not demonstrate a change in membrane fluidity with aging. Compared with those in young rat ECs, cholesterol concentrations in aged rat ECs were significantly higher, whereas phospholipid concentrations were unchanged; consequently, the cholesterol/phospholipid molar ratio was significantly higher in aged rat ECs. Lipid peroxide levels measured with thiobarbituric acid reactive substances were significantly higher in EC plasma membranes of aged rats. These results indicate that age-related increases in cholesterol and lipid peroxide in vascular EC plasma membranes reduce membrane fluidity.