Effect of cholesterol on the lactosylceramide domains in phospholipid bilayers

Lactosylceramide (LacCer) in the plasma membranes of immune cells is an important lipid for signaling in innate immunity through the formation of LacCer-rich domains together with cholesterol (Cho). However, the properties of the LacCer domains formed in multicomponent membranes remain unclear. In this study, we examined the properties of the LacCer domains formed in Cho-containing 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) membranes by deuterium solid-state NMR and fluorescence lifetimes. The potent affinity of LacCer-LacCer (homophilic interaction) is known to induce a thermally stable gel phase in the unitary LacCer bilayer. In LacCer/Cho binary membranes, Cho gradually destabilized the LacCer gel phase to form the liquid-ordered phase by its potent order effect. In the LacCer/POPC binary systems without Cho, the ²H NMR spectra of 10′,10′-d₂-LacCer and 18′,18′,18′-d₃-LacCer probes revealed that LacCer was poorly miscible with POPC in the membranes and formed stable gel phases without being distributed in the liquid crystalline domain. The lamellar structure of the LacCer/POPC membrane was gradually disrupted at around 60°C, whereas the addition of Cho increased the thermal stability of the lamellarity. Furthermore, the area of the LacCer gel phase and its chain order were decreased in the LacCer/POPC/Cho ternary membranes, whereas the liquid-ordered domain, which was observed in the LacCer/Cho binary membrane, was not observed. Cho surrounding the LacCer gel domain liberated LacCer and facilitated forming the submicron to nano-scale small domains in the liquid crystalline domain of the LacCer/POPC/Cho membranes, as revealed by the fluorescence lifetimes of trans-parinaric acid and trans-parinaric acid-LacCer. Our findings on the membrane properties of the LacCer domains, particularly in the presence of Cho, would help elucidate the properties of the LacCer domains in biological membranes.     

Effect of dimethylaminoethylphosphonate on phospholipid metabolism in Tetrahymena

Dimethylaminoethylphosphonate (DMAEP) was incorporated into the phospholipids of the ciliate protozoan Tetrahymena thermophila at the expense of both phosphatidylethanolamine and phosphatidylcholine, but it had no effect on the levels of the 2-aminoethylphosphonolipid. The newly formed DMAEP-lipid accounted for almost 50% of the phospholipids of the organism. The DMAEP was incorporated into the phospholipids using both the ethanolaminephosphotransferase and cholinephosphotransferase pathways. The DMAEP-lipid was not methylated to the trimethyl derivative, confirming the lack of methylation of phosphonolipids by Tetrahymena.     

Effect of mevinolin and glycyrrhizinic acid on cholesterol and bile acid metabolism in cultured rabbit hepatocytes]

A comparison of effects of two hypocholesterolemic drugs--mevinolin and glycyrrhizinic acid, on cholesterol and bile acid metabolism in cultured rabbit hepatocytes has been carried out. The following parameters have been determined: i) cholesterol synthesis from [2-14C]acetate; ii) bile acid production from newly synthesized and [4-14C]-labeled HDL2 cholesterol, and, iii) total cholesterol efflux into the incubation medium Mevinolin (0.5 microgram/ml) inhibited [2-14C] acetate incorporation into cholesterol by more than 90%. Conversely, glycyrrhizinic acid did not influence cholesterol synthesis even when used at high (100 micrograms/ml) concentrations but stimulated the conversion of endogenous (by 37%) and exogenous (by 18%) cholesterol into bile acids and increased, in addition, the proportion of bile acids in the total sterol pool released from hepatocytes into the incubation medium. At the same time, mevinolin used at 0.5 microgram/ml decreased the bile acid production by endogenous (by 27%) and exogenous (by 40%) cholesterol. The data obtained suggest that glycyrrhizinic acid exerts hypocholesterolemic action by stimulation of cholesterol conversion into bile acids without any effect on cholesterol synthesis. As for mevinolin, it has a cholesterol-suppressing effect via a mechanism of cholesterol synthesis inhibition only.     

Effect of ubiquinone on phospholipid metabolism in radiation injury

The synthesis and phospholipid content in the liver, intestine and spleen in normal and irradiated rats administered ubiquinone-9 were studied with the use of 3H-serine. Ubiquinone markedly activated decarboxylation of phosphatidylserine and suppressed transformation of phosphatidylethanolamine to phosphatidylcholine in rat liver and spleen. The effect was also observed in the organs of irradiated animals. In rat intestine, administration of ubiquinone normalized a sharp gamma-irradiation-induced inhibition of transformation of phosphatidylcholine from phosphatidylethanolamine. The catabolism of phospholipids under the action of ubiquinone and radiation was inhibited in the liver and, on the contrary, was activated in radiosensitive organs.     

Effect of age and strain on cerebral phospholipid metabolism

It has been demonstrated that spontaneously hypertensive adult rats (SHR) develop severe hypertension and cerebrovascular lesions on drinking 1% NaCl from weaning. Phospholipid metabolism is actively altered in these severely lesioned animals (SHR-NaCl) as compared to SHRs which drink only water and showed only sporadic cerebrovascular lesions. We have tested the incorporation of water soluble phospholipid precursors into the corresponding phospholipid from different brain areas, by injecting either a mixture of labeled glycerol and choline or glycerol and ethanolamine into the lateral ventricle of the brain of adult (4 months old) and senescent (12 months old) SHR-NaCl. The results were compared to those obtained from 4 and 12 months old Wistar normotensive rats. When adult normotensive rats were compared with adult hypertensive rats (4-SHR-NaCl) incorporation was found to decrease in some areas according to the precursors injected. Similar results were obtained from 12 month old normotensive Wistar rats that, however, showed a decrease in phospholipid biosynthesis in all the area tested. Interestingly, no significant differences of incorporation rate were found between 12 month old normotensive and 12 month old hypertensive rats.     

Effect of free cholesterol on incorporation of triolein in phospholipid bilayers

Triacylglycerols are the major substrates for lipolytic enzymes that act at the surface of emulsion-like particles such as triglyceride-rich lipoproteins, chylomicrons, and intracellular lipid droplets. This study examines the effect of cholesterol on the solubility of a triacylglycerol, triolein, in phospholipid surfaces. Solubilities of [carbonyl-13C]triolein in phospholipid bilayer vesicles containing between 0 and 50 mol % free cholesterol, prepared by cosonication, were measured by 13C NMR. The carbonyl resonances from bilayer-incorporated triglyceride were shifted downfield in the 13C NMR spectra from those corresponding to excess, nonincorporated material. This enabled solubilities to be determined directly from carbonyl peak intensities at most cholesterol concentrations. The bilayer solubility of triolein was inversely proportional to the cholesterol/phospholipid mole ratio. In pure phospholipid vesicles the triolein solubility was 2.2 mol %. The triglyceride incorporation decreased to 1.1 mol % at a cholesterol/phospholipid mole ratio of 0.5, and at a mole ratio of 1.0 for the bilayer lipids, the triolein solubility was reduced to just 0.15 mol %. The effects of free cholesterol were more pronounced and progressive than observed previously on the bilayer solubility of cholesteryl oleate (Spooner, P. J. R., Hamilton, J. A., Gantz, D. L., & Small, D. M. (1986) Biochim. Biophys. Acta 860, 345-353]. As with cholesteryl oleate, we suggest that cholesterol also displaces solubilized triglyceride to deeper regions of the bilayer.     

Effect of ethanol on cholesterol and phospholipid composition of HeLa cells

Chronic exposure of animals to ethanol leads to changes in membrane lipid composition which may be related to the development of tolerance and physical dependence. The object of the present study was to investigate this phenomenon at a cellular level. HeLa cells were grown in the presence of ethanol (86 mM) for periods of up to 9 days. Both the cholesterol and phospholipid concentration of these cells increased during this period but the cholesterol:phospholipid ratio remained unchanged. Among the phospholipid classes phosphatidic acid decreased while phosphatidylethanolamine, phosphatidylcholine and phosphatidylserine increased rapidly, returning toward control values by 9 days. Significant decreases were observed in saturated (14:0, 16:0) and monoenoic (16:1, 18:1) fatty acids while the major polyenoic fatty acid (20:4) increased. It is concluded that cultured mammalian cells represent a useful model for investigation of the direct effects of ethanol on membrane lipid metabolism.     

Effect of cholesterol and surfactant protein B on the viscosity of phospholipid mixtures

Low viscosity of the surface of alveolar fluid is mandatory for undisturbed surfactant function. Based on the known reduction of the viscosity of surfactant-like phospholipid (PL-) mixtures by plasmalogens, the effect of cholesterol and surfactant protein (SP-) B on surface viscosity of these lipid mixtures has been studied. Surface viscosity at the corresponding surface tension was measured with the oscillating drop surfactometer. We found that the viscosity was lowest in cholesterol-, followed by plasmalogen- and SP-B containing samples. Addition of SP-B to a plasmalogen-containing PL-mixture caused a further decrease in viscosity. However, in cholesterol containing mixtures, addition of SP-B led to a significant increase in viscosity, and the effect was reversed by further addition of plasmalogens. We conclude that SP-B, plasmalogens and cholesterol all affect the surface viscosity, thus synergistically regulate monolayer stability. This suggests that they are all needed in vivo for fine tuning of surface properties of pulmonary surfactant.     

Effect of cholesterol on the structure and dynamic properties of unsaturated phospholipid bilayers

Molecular dynamics (MD) computer simulations of five different hydrated unsaturated phosphatidylcholine lipid bilayers built up by 18:0/18:1(n-9)cis PC, 18:0/18:2(n-6)cis PC, 18:0/18:3(n-3)cis PC, 18:0/20:4(n-6)cis PC, and 18:0/22:6(n-3)cis PC molecules with 40 mol% cholesterol, and the same five pure phosphatidylcholine bilayers have been performed at 303 K. The simulation box of a lipid bilayer contained 96 phosphatidylcholines, 64 cholesterols, and 3840 water molecules (48 phosphatidylcholine molecules and 32 cholesterols per layer and 24 water molecules per phospholipid or cholesterol in each case). The lateral self-diffusion coefficients of the lipids in these systems and mass density profiles with respect to the bilayer normal have been analyzed. It has been found that the lateral diffusion coefficients of phosphatidylcholine molecules increase with increasing number of double bonds in one of the lipid chains, both in pure bilayers and in bilayers with cholesterol. It has been found as well that the lateral diffusion coefficient of phosphatidylcholine molecules of a lipid bilayer with 40 mol% cholesterol is smaller than that for the corresponding pure phosphatidylcholine bilayer.     

Effect of dietary tomatine on cholesterol metabolism in the rat

Tomatine is a virtually nonabsorbable saponin which has been used as an antifungal agent and analytically as a cholesterol precipitant. It was used in this study to determine whether or not it can form a complex with cholesterol in vivo in the rat intestine and what effects such complex formation would have on cholesterol metabolism. Rats that were fed tomatine as 1% of the diet had a decreased uptake of dietary cholesterol by the liver, an increased rate of hepatic and intestinal cholesterol synthesis as well as a partial offsetting of the dietary cholesterol-induced decrease in hepatic cholesterogenesis, and an apparent increase in sterol excretion without an effect on bile acid excretion. In vitro, tomatine did not sequester cholic acid as did cholestyramine. The results show that tomatine has an effect on cholesterol absorption and on other aspects of lipid metabolism in the rat similar to that of cholestyramine, with the notable exception that tomatine increased sterol excretion while cholestyramine increased bile acid excretion. It was suggested that tomatine forms a nonabsorbable complex with cholesterol in the rat intestine.     

Effect of dietary cholesterol and alloxan-diabetes on tissue cholesterol and apolipoprotein E mRNA levels in the rabbit

Alloxan-diabetic rabbits develop hypercholesterolemia and hypertriglyceridemia in response to cholesterol feeding. To determine whether alterations in apolipoprotein composition of plasma lipoproteins were due to changes in apolipoprotein gene expression, we measured the steady state apoE mRNA levels in several tissues from both control and diabetic rabbits. Control rabbits were fed either chow or chow plus 1.5% cholesterol (chow-fed or cholesterol-fed groups) and diabetic rabbits were fed either chow or chow plus 0.5% cholesterol for dietary periods of 5, 21, and 42 days. The tissues examined were liver, small intestine, brain, adrenals, and aorta. ApoE mRNA levels were measured by Northern and dot blot analysis with a human apoE cDNA probe. In control rabbits fed either chow or cholesterol diets for up to 42 days, the steady state apoE mRNA levels remained relatively constant in all of the tissues examined. In contrast, in alloxan-diabetic rabbits fed a 0.5% cholesterol diet, apoE mRNA was reduced in liver, brain, and adrenals (46 +/- 19%, 56 +/- 5%, and 39 +/- 18% of chow-fed control, respectively), but showed little change in the aorta (91 +/- 22% of chow-fed control). Despite a similar increase in plasma cholesterol, the cholesterol content of the liver and adrenals of cholesterol-fed diabetic rabbits were 20% and 50%, respectively, of that of the liver and adrenals in cholesterol-fed control rabbits. The result that apoE mRNA levels and tissue cholesterol content are altered in the diabetic cholesterol-fed rabbit suggests that insulin deficiency in the rabbit may influence apoE gene expression and tissue cholesterol homeostasis.     

Effect of hormones on phospholipid metabolism in human cultured fibroblasts

The effect of hormones on phospholipid metabolism, pool size, 32P labeling and changes in fatty acid of human adult fibroblasts was determined. Simultaneously the change in membrane fluidity of single cells was recorded via fluorescence recovery after photobleaching under the influence of hormones. From all substances tested (isoproterenol, phenylephrine, adrenalin, histamine, angiotensin II, dansylcadaverine, propranolol) only isoproterenol and adrenalin slightly decreased total amount of phosphatidylcholine (PC). The amount of the other phospholipids analyzed remained unchanged. The 32P incorporation rate into phospholipids (PC, phosphatidylinositol (PI), phosphatidylethanolamine (PE)) was affected basicly different analyzing either PC, PI or PE. Histamine and propranolol provoked the highest incorporation of 32P (240% increase in PI labeling). Isoproterenol and adrenalin decreased PC labeling (45% and 18%) whereas isoproterenol decreased 32P incorporation into PI (18%), and adrenalin led to an increase (37%). PE labeling showed no or a slight increase in 32P incorporation applying the other agonists or antagonists. The fatty acid pattern of the respective phospholipids changed only to a minor extend. A decrease in hexadecanoic acid content of PI was found after administration of either isoproterenol, adrenalin or histamine. Parallel determination of membrane fluidity of single cells by fluorescence recovery after photobleaching showed an increase in the diffusion coefficient of a fluorescent lipid probe sticking in the membrane, following administration of isoproterenol and adrenalin, other substances tested exerted no effect. A relationship to changes in phospholipid metabolism became obvious. These results are discussed considering known mechanisms of receptor coupling and change in phospholipid metabolism and fluidity.