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Effect of heparin on lecithin: cholesterol acyltransferase and lipids in cholestasis.

The effect of heparin on the proportion of cholesterol esters, lecithin/lysolecithin ratio, and lecithin: cholesterol acyltransferase (LCAT) activity was studied in twelve patients with cholestasis. A good correlation was observed between a decreased LCAT activity and the proportion of cholesterol esters, and no consistent variation was noted after heparin. The lecithin/lysolecithin ratio was increased in 7 patients and became normal after heparin in 6 cases. The clearance of lipoprotein-X after heparin seems to be not related to LCAT activity. These results demonstrate that heparin does not modify the LCAT activity, and suggest that heparin induces an increase of phospholipase activity in some patients with cholestasis.

Acyltransferases↗

Serum lecithin: cholesterol acyltransferase activity in the bile duct-ligated rat.

Serum lipoprotein cholesterol concentration, lecithin: cholesterol acyltransferase (LCAT) activity and lipoprotein X (LP-X) were studied after acute cholestasis had been produced by ligation of the common bile duct in rats fed a liquid fat-free diet. The concentration of serum-free cholesterol, predominantly contained in the low density lipoprotein fraction, increased steadily up to 72 h after ligation. The concentration of cholesterol in the high density lipoprotein fraction did not change significantly, and similarly the serum concentration of esterified cholesterol was not altered. The activity of LCAT, expressed in molar terms as millimoles of cholesterol esterified per litre per hour, did not change during the 72-hour study period. These data indicate acute cholestasis in the rat does not result in a LCAT deficiency and suggest that the accumulation of serum-free cholesterol in this situation is independent of the serum cholesterol-esterifying mechanism.

Acute Disease↗

Trans fatty acids: transport and positional specificity in rat placental lecithin.

Differences in positional incorporation of trans and cis isomers of octadecenoic and octadecadienoic acids in placental lecithin of rats was demonstrated. A 14C-labeled albumin complex of elaidic, oleic, linoelaidic, or linoleic acid was injected into the jugular vein of pregnant rats. 6 h later 45-64% of the total radioactivity in placental lipids was found in phospholipids (PL), with a major portion of the label incorporated into choline phosphoglycerides (CPC). Following hydrolysis of placental CPC by phospholipase A2, distribution of radioactivity within isolated fatty acids and lysolecithin suggested preferential incorporation of t,t-18:2 at position 2 and a nearly equal distribution of t-18:1 at positions 1 and 2.

Animals↗

Lecithin-cholesterol acyltransferase activity in children in the early neonatal period.

Changes in the rate of plasma cholesterol esterification occurring shortly after birth were followed by determining the activity of lecithin-cholesterol acyltransferase (EC 2.3.1.43) in newborn children, and at the age of 3 days and 3 months. The radioassay method using 'common' substrate was applied. The molar esterification rate (MER) in umbilical cord serum was about half of the values found in the sera of 3-day-old and 3-month-old children, or adult subjects. Because of a positive correlation between MER and the plasma concentration of unesterified cholesterol, the values of fractional esterification rate in children were in good agreement with those characteristic for healthy adults.

Adult↗

Apolipoprotein A-I-containing lipoproteins in human umbilical cord blood. Relation to proapolipoprotein A-I and lecithin:cholesterol acyltransferase.

Lipids, apolipoproteins, lipoproteins, as well as lipoproteins containing both apo A-I and apo A-II (Lp A-I:A-II) or apo A-I but no apo A-II (Lp A-I), proapolipoprotein (proapo) A-I and the activity of lecithin:cholesterol acyltransferase (LCAT), were investigated in umbilical cord sera of 67 term human neonates (30 females and 37 males). Lp A-I and Lp A-I:A-II were present in umbilical cord sera with levels of 0.26 +/- 0.1 and 0.33 +/- 0.15 g/l, respectively. Furthermore, the absolute amount of proapo A-I was lower in cord blood than in adult plasma, but in view of the lower apo A-I levels in umbilical cord sera it comprised 10.48 +/- 3.86% of total apo A-I and was thus significantly higher than in adult plasma (7.1 +/- 0.9%). Proapo A-I was highly correlated with HDL cholesterol and apo A-I. Total serum LCAT activity was about 50% of adult plasma and was highly correlated with Lp A-I, but not with Lp A-I:A-II. We conclude that human umbilical cord serum contains both Lp A-I and Lp A-I:A-II particles and that the LCAT activity is predominantly related with the Lp A-I subfraction. The higher percentage in umbilical cord sera of proapo A-I may indicate a higher turnover of apo A-I or a lower activity of the proapo A-I cleaving enzyme which is still not identified.

Apolipoprotein A-I↗

Postnatal development of plasma-lipid-clearing enzymes (lipoprotein lipase, hepatic lipase and lecithin:cholesterol acyl transferase) and lipid profiles in suckling rats.

We examined the activity of plasma circulating lipoprotein lipase and hepatic lipase, lecithin:cholesterol acyl transferase (LCAT), as well as lipid profiles in Sprague-Dawley rats from 1 day until 29 days of age. Plasma lipoprotein lipase activity peaked between ages 5 and 15 days and decreased after weaning, while plasma hepatic lipase activity remained constantly low during the suckling period and increased after weaning. No statistically significant difference in LCAT activity was seen from birth until weaning. Plasma triglycerides, as well as free fatty acids, decreased significantly after birth. Total plasma cholesterol increased during the suckling period and decreased after weaning. HDL cholesterol increased after the first 10 days of life, and free cholesterol remained constant after an initial decrease from birth to the 5th day of life. In conclusion, the enzymes associated with the metabolism of triglycerides, cholesterols and phospholipids are well developed in the rat shortly after birth.

Animals↗

Changes in fatty acid composition of cervical mucus lecithin during pregnancy.

The fatty acid composition in cervical mucus was determined in 37 pregnant and 17 nonpregnant women using gas liquid chromatography. In both groups palmitic acid (16:0) and oleic acid (18:1) were the predominant acids comprising more than half of the total amount. Compared to nonpregnant controls, pregnant women had markedly elevated levels of oleic acid (18:1), while mean levels of myristic acid (14:0) and stearic acid (18:0) were significantly lower. These pregnancy-induced changes in fatty acid pattern could not be ascribed to the increased viscosity of cervical mucus. Only minor differences in fatty acid pattern were found between women in first trimester and at term. In contrast to nonpregnant controls, a significant negative correlation between levels of myristic acid (14:0) and docosahexaenoic acid (22:6) was found in pregnant women, indicating a redistribution of these two fatty acids during pregnancy. The present study demonstrates that pregnancy alters the fatty acid composition of cervical mucus lecithin, and that the characteristic changes are present before the 10th gestational week.

Adult↗

Lecithin:cholesterol acyltransferase activity and HDL composition in serum of patients with kidney transplants.

In comparison with a control group, lecithin:cholesterol acyltransferase activities and HDL-cholesterol concentrations in serum of patients with kidney transplants were unchanged, while apolipoprotein A and HDL-triglyceride concentrations were increased. This constellation shows that the cholesterol metabolism in patients with kidney transplants differs from that of patients undergoing chronic haemodialysis.

Apolipoproteins↗

Changes in serum lipids and lecithin: cholesterol acyltransferase enzyme during 1 week weight reduction of women on a low-calorie diet.

In 32 women of normal body weight who volunteered to participate in the study, the effect of rapid weight reduction by a low-calorie liquid diet on serum lipids and lecithin:cholesterol acyltransferase (LCAT) enzyme activity was studied. Women were on this 400 kJ/day diet for 7 days and fasting blood samples were drawn before and immediately after the diet. Serum cholesterol decreased from 5.7 +/- 1.0 to 5.2 +/- 1.1 mmol/l and high density lipoprotein cholesterol from 1.77 +/- 0.43 to 1.50 +/- 0.35 mmol/l. The serum LCAT activity decreased significantly during the weight reduction period. When serum LCAT activity was correlated to lipid parameters, a positive correlation was found with total cholesterol and triglyceride concentrations before weight reduction and also between changes in LCAT activity and total cholesterol concentration. The data suggest that serum LCAT activity might have a prominent role in the regulation of serum lipid levels.

Acyltransferases↗

Cholesteryl ester transfer protein expressed in lecithin cholesterol acyltransferase-deficient mice.

OBJECTIVE: Regulation of plasma cholesteryl ester transfer protein (CETP) concentration was studied in lecithin-cholesterol acyltransferase (LCAT)-knockout mice. METHODS AND RESULTS: LCAT-knockout mice were cross-bred with CETP transgenic mice. The offspring (n=63) were classified for LCAT genotype and plasma CETP levels (no CETP, low CETP, and high CETP). High density lipoprotein (HDL) decreased as LCAT decreased in each CETP-level group. In the lcat(+/+) and lcat(+/-) mice, plasma CETP varied from 0 to 30 micro g/mL, whereas it was <10 micro g/mL in the lcat(-/-) mice. HDL cholesterol and phospholipid decreased and HDL triglyceride and apolipoprotein B increased in CETP in the lcat(+/+) and lcat(+/-) mice, whereas there was no difference in HDL between low and high CETP. An effect of CETP on HDL was not detected in the lcat(-/-) mice because of the absence of mature HDL. Genomic DNA and mRNA of CETP were correlated and were similar in the lcat(-/-) and lcat(+/+) mice. Plasma CETP was correlated with its genomic DNA and mRNA, but the slope of the increase was much lower in the lcat(-/-) mice. Whereas plasma CETP mostly associates with HDL in the lcat(+/+) mouse, it is found free in the lcat(-/-) mouse. CONCLUSIONS: Plasma CETP is posttranscriptionally downregulated in the lcat(-/-) mice, presumably by its extremely low HDL.

Animals↗

Effect of fish oil on lipoproteins, lecithin:cholesterol acyltransferase, and lipid transfer protein activity in humans.

A group of 33 mildly hypercholesterolemic men were stratified into three groups on diets closely matched except for the polyunsaturated fatty acid supplement. The first group received 14 g/day of linoleic acid (safflower oil); the second group, 9 g of alpha-linolenic acid (linseed oil); and the third group, 3.8 g of n-3 fatty acids (fish oil). Only fish oil lowered plasma triglycerides (by 24% at 6 weeks, p less than 0.05 compared to safflower oil). Very low density lipoprotein (VLDL) apoprotein (apo) B, triglyceride, and cholesterol all fell significantly with the fish-oil diet (p less than 0.01). Low density lipoprotein (LDL) cholesterol fell by 0.18 and 0.10 mmol/l, respectively, with the safflower-oil and linseed-oil diets, but rose by 0.24 mmol/l with the fish-oil diet (p less than 0.05). There was a strong correlation between the changes in VLDL triglyceride and LDL cholesterol with the fish-oil diet (r = -0.84, p less than 0.002). High density lipoprotein (HDL) cholesterol fell slightly in all three groups (p less than 0.02 with the linseed-oil diet only). However, the apo A-I/A-II ratio rose by 5% (p less than 0.05), and the HDL2/HDL3 protein ratio increased by 28% with the fish-oil diet (p less than 0.005). Fish oil reduced the capacity for transfer of cholesteryl ester between LDL and HDL by 23% (p less than 0.02 compared to baseline), reduced plasma lecithin:cholesterol acyltransferase activity by 21% (p less than 0.05), and reduced maximal stimulated thromboxane production by 9% (p less than 0.05). Thus fish oil produced three potentially beneficial changes: significant decreases in VLDL concentration and in thromboxane production and an increase in the HDL2/HDL3 ratio. The increase in the average HDL particle size probably reflected reduced cholesteryl ester acceptor capacity within the smaller pool of VLDL, as well as the decline in lipid transfer activity in plasma involving transfer protein itself, LDL, and HDL.

Apolipoproteins↗

Use of synthetic peptide analogues to localize lecithin:cholesterol acyltransferase activating domain in apolipoprotein A-I.

The major protein of high density lipoprotein (HDL), apolipoprotein (apo) A-I, is the major activator of the plasma enzyme lecithin:cholesterol acyltransferase (LCAT). A consensus amino acid sequence has been defined for the eight, 22-residue long, tandem amphipathic helical repeats located in the carboxy-terminal region of apo A-I. A series of 22 and 44mer synthetic peptide analogues of the consensus domain, differing only in their 13th amino acid residue, were prepared and tested for LCAT activation. One of the peptides was found to equal apo A-I in LCAT activation. This is the first time a peptide activator for LCAT that rivals the activity of apo A-I in the vesicular and discoidal egg phosphatidylcholine assay systems has been synthesized. Based on these results, we propose that the major LCAT-activating domain of apo A-I resides in the 22mer tandem repeats, each containing Glu at the 13th residue and located between residues 66 and 121 in the native apolipoprotein.

Amino Acid Sequence↗

Inhibition of lecithin-cholesterol acyltransferase and modification of HDL apolipoproteins by aldehydes.

Experimental evidence suggests that aldehydes generated as a consequence of lipid peroxidation may be involved in the pathogenesis of atherosclerosis. It is well documented that aldehydes modify LDL: however, less is known concerning the effects of aldehydes on other plasma and interstitial fluid components. In the present study, we investigated the effects of five physiologically relevant aldehydes (acetaldehyde, acrolein, hexanal, 4-hydroxynonenal [HNE], and malondialdehyde [MDA]) on two key constituents of the antiatherogenic reverse cholesterol transport pathway, lecithin-cholesterol acyltransferase (LCAT) and HDL. Human plasma was incubated for 3 hours at 37 degrees C with each one of the five aldehydes at concentrations ranging from 0.16 to 84 mmol/L. Dose-dependent decreases in LCAT activity were observed. The short-chain (acrolein) and long-chain (HNE) alpha,beta-unsaturated aldehydes were the most effective LCAT inhibitors. Micromolar concentrations of these unsaturated aldehydes resulted in significant reductions in plasma LCAT activity. The short- and longer-chain saturated aldehydes acetaldehyde and hexanal and the dialdehyde MDA were considerably less effective at inhibiting LCAT than were acrolein and HNE. In addition to inhibiting LCAT, aldehydes increased HDL electrophoretic mobility and cross-linked HDL apolipoproteins. Cross-linking of apolipoproteins A-I and A-II required higher aldehyde concentrations than inhibition of LCAT. The alpha,beta-unsaturated aldehydes acrolein and HNE were fourfold to eightfold more effective cross-linkers of apolipoproteins A-I and A-II than the other aldehydes studied. These data suggest that products of lipid peroxidation, especially unsaturated aldehydes, may interfere with normal HDL cholesterol transport by inhibiting LCAT and modifying HDL apolipoproteins.

Aldehydes↗

Determinants of plasma HDL-cholesterol in hypertriglyceridemic patients. Role of cholesterol-ester transfer protein and lecithin cholesteryl acyl transferase.

Hypertriglyceridemic patients commonly have low levels of HDL cholesterol. Elevated triglycerides per se may be one cause of low HDL levels, but other factors also may be involved. The current study was designed to define the role of cholesterol-ester transfer protein (CETP) in causation of a low HDL cholesterol in hypertriglyceridemic patients; in addition other factors-lecithin cholesterol acyl transferase (LCAT), hepatic triglyceride lipase (HTGL), and lipoprotein lipase (LPL)-were examined. Plasma activities of CETP and LCAT were measured in 137 male patients with moderate hypertriglyceridemia (plasma triglycerides [TGs] 200 to 500 mg/dL and LDL cholesterol < 160 mg/dL). Results were compared with those from 50 normolipidemic men of similar age and body habitus. In addition, lipase activities in postheparin plasma were measured in 118 of the subjects with hypertriglyceridemia. The activities of CETP and LCAT were 17% (P < .01) and 7% (P < .05), respectively, higher in the hypertriglyceridemic group than in control subjects. By stepwise regression analysis CETP appeared to contribute 15.2% and LCAT 9.8% to variation in HDL-cholesterol levels. Activities of LPL and HTGL together contributed an additional 14.1% to HDL-cholesterol variation. In contrast, levels of plasma TG accounted for only 5.4% of the variation. There were no differences in relative contributions of these parameters in patients with and those without coronary heart disease. This study indicates that several factors contribute to the variation in HDL-cholesterol levels in hypertriglyceridemic patients, and five factors-CETP, LCAT, HTGL, LPL, and triglyceride levels-account for almost half of this variation.

Aged↗

Apolipoprotein A-IFIN (Leu159-->Arg) mutation affects lecithin cholesterol acyltransferase activation and subclass distribution of HDL but not cholesterol efflux from fibroblasts.

We showed earlier that the apolipoprotein A-I Leu159-->Arg mutation (apoA-IFin) results in dominantly inherited hypoalphalipoproteinemia. In the present study we investigated the effect of the apoA-IFin mutation on lipoprotein profile, apoA-I kinetics, lecithin:cholesterol acyltransferase (LCAT) activation, and cholesterol efflux in vitro. Carriers (n = 9) of the apoA-IFin mutation exhibited several lipoprotein abnormalities. The serum HDL cholesterol level was diminished to 20% of normal, and nondenaturing gradient gel electrophoresis of HDL showed disappearance of particles at the 9.0- to 12-nm size range (HDL2-type) and the presence of small 7.8- to 8.9-nm (mostly HDL3-type) particles only. HDL3-type particles from both the mutation carriers and nonaffected family members were similarly converted to large, HDL2-type particles by phospholipid transfer protein in vitro. Studies on apoA-I kinetics in four affected subjects favored accelerated catabolism of apoA-I. Experiments with reconstituted proteoliposomes showed that the capacity of apoA-IFin protein to activate LCAT was reduced to 40% of that of the wild-type apoA-I. The impact of the apoA-IFin protein on cholesterol efflux was examined in vitro using [3H]cholesterol-loaded human fibroblasts and three different cholesterol acceptors: (1) total HDL, (2) total apoA-I combined with phospholipid, and (3) apoA-I isoform (apoA-IFin or wild-type apoA-I isoform 1) combined with phospholipid. ApoA-IFin did not impair phospholipid binding or cholesterol efflux from fibroblasts to any of the acceptors used. Only one of the nine apoA-IFin carriers appears to have evidence of clinically manifested atherosclerosis. In conclusion, although the apoA-IFin mutation does not alter the properties of apoA-I involved in promotion of cholesterol efflux, its ability to activate LCAT in vitro is defective. In vivo, apoA-IFin was found to be associated with several lipoprotein composition rearrangements and increased catabolism of apoA-I.

Adolescent↗

The Arg123-Tyr166 central domain of human ApoAI is critical for lecithin:cholesterol acyltransferase-induced hyperalphalipoproteinemia and HDL remodeling in transgenic mice.

High density lipoprotein (HDL) metabolism and lecithin:cholesterol acyltransferase (LCAT)-induced HDL remodeling were investigated in transgenic mice expressing human apolipoprotein (apo) AI or an apoAI/apoAII chimera in which the Arg123-Tyr166 domain of apoAI was substituted with the Ser12-Ala75 domain of apoAII. Expression of apoAI and of the apoAI/apoAII chimera resulted in a respective 3. 5-fold and 2.9-fold increase of HDL cholesterol. Human LCAT gene transfer into apoAI-transgenic mice resulted in a 5.1-fold increase of endogenous LCAT activity. This increase was associated with a 2. 4-fold increase of the cholesterol ester-to-free cholesterol ratio of HDL, a shift from HDL(3) to HDL(2), and a 2.4-fold increase of HDL cholesterol levels. Agarose gel electrophoresis revealed that human LCAT gene transfer into human apoAI-transgenic mice resulted in an increase of pre-beta-HDL and of pre-alpha-HDL. In contrast, human LCAT gene transfer did not affect cholesterol levels and HDL distribution profile in mice expressing the apoAI/apoAII chimera. Mouse LCAT did not "see" a difference between wild-type and mutant human apoAI, whereas human LCAT did, thus localizing the species-specific interaction in the central domain of apoAI. In conclusion, the Arg123-Tyr166 central domain of apoAI is not critical for in vivo lipoprotein association. It is, however, critical for LCAT-induced hyperalphalipoproteinemia and HDL remodeling independent of the lipid-binding properties of apoAI.

Amino Acid Sequence↗

Ldl modified by hypochlorous acid is a potent inhibitor of lecithin-cholesterol acyltransferase activity.

Modification of low density lipoprotein (LDL) by myeloperoxidase-generated HOCl has been implicated in human atherosclerosis. Incubation of LDL with HOCl generates several reactive intermediates, primarily N-chloramines, which may react with other biomolecules. In this study, we investigated the effects of HOCl-modified LDL on the activity of lecithin-cholesterol acyltransferase (LCAT), an enzyme essential for high density lipoprotein maturation and the antiatherogenic reverse cholesterol transport pathway. We exposed human LDL (0.5 mg protein/mL) to physiological concentrations of HOCl (25 to 200 micromol/L) and characterized the resulting LDL modifications to apolipoprotein B and lipids; the modified LDL was subsequently incubated with apolipoprotein B-depleted plasma (density >1.063 g/mL fraction), which contains functional LCAT. Increasing concentrations of HOCl caused various modifications to LDL, primarily, loss of lysine residues and increases in N-chloramines and electrophoretic mobility, whereas lipid hydroperoxides were only minor products. LCAT activity was extremely sensitive to HOCl-modified LDL and was reduced by 23% and 93% by LDL preincubated with 25 and 100 micromol/L HOCl, respectively. Addition of 200 micromol/L ascorbate or N-acetyl derivatives of cysteine or methionine completely prevented LCAT inactivation by LDL preincubated with </=200 micromol/L HOCl. Protecting the free thiol groups of LCAT with 5,5'-dithio-bis-(2-nitrobenzoic acid) before exposure to HOCl-modified LDL, which inhibits lipid hydroperoxide-mediated inactivation of LCAT, failed to prevent the loss of enzyme activity. Our data indicate that N-chloramines from HOCl-modified LDL mediate the loss of plasma LCAT activity and provide a novel mechanism by which myeloperoxidase-generated HOCl may promote atherogenesis.

Chloramines↗

Epidemiological correlates of high density lipoprotein subfractions, apolipoproteins A-I, A-II, and D, and lecithin cholesterol acyltransferase. Effects of smoking, alcohol, and adiposity.

Recent data suggest that the protection against ischemic heart disease afforded by high density lipoprotein (HDL) cholesterol (C) may be concentrated in the HDL2 subfraction. To examine the behavioral correlates of the HDL subfractions, we recalled 33 men and 17 women of a random sample from the Pacific Northwest Bell Telephone Company Health Survey. Adiposity and very low density lipoprotein (VLDL) triglyceride were negatively correlated with HDL2C. Smoking was not correlated with HDL2C, but was negatively correlated with HDL3C (men, rs = -0.635, p = 0.001; women, rs = -0.534, p = 0.014); this relationship was independent of alcohol consumption, adiposity, and VLDL triglyceride. Alcohol consumption was also more strongly related to HDL3C (men, rs = 0.248, p = 0.082; women, rs = 0.586, p = 0.007). Lecithin cholesterol acyltransferase (LCAT) mass was negatively related with HDL2C, but was positively correlated with HDL3C and apolipoprotein A-II. Smoking was negatively correlated with LCAT mass. Since it is believed that HDL3C is not associated with the risk of ischemic heart disease and since both smoking and alcohol consumption may mainly affect HDL3C, the current study suggests that the increase in risk of ischemic heart disease with smoking and the possible decrease with alcohol consumption may be mediated through mechanisms other than their effects on HDLC.

Adult↗