Plasma Low-density Lipoprotein Concentrations Research Articles

In vitro Characterization of a Membrane-Based Low-Density Lipoprotein Affinity Adsorption Device

The objective of this study was to explore the use of microporous membranes as an alternative substrate to porous beads in affinity adsorption of low-density lipoprotein (LDL) for therapeutic purposes. Flat sheet immunoaffinity membranes containing a polyclonal antibody preparation were utilized as the affinity substrate. The antibody was covalently immobilized to the surface through a poly(ethylene glycol) (PEG) spacer. Equilibrium adsorption of LDL from plasma was measured. Adsorption from plasma and elution of bound LDL using citrate buffer were studied as a function of flow rate.Specific capacity was as great as 2 mg apolipoprotein B per milliliter membrane volume. The superior transport properties of the membrane allowed rapid adsorption and regeneration, which translated to a large number of adsorptive cycles that can be performed within a given treatment time. On the basis of in vitro performance characteristics, it is estimated that an immunoaffinity membrane device can provide a reduction in patient plasma LDL concentration comparable to that provided by packed columns, but with almost an 80% reduction in the device volume.

Overexpression of Cholesterol 7α-Hydroxylase (CYP7A) in Mice Lacking the Low Density Lipoprotein (LDL) Receptor Gene: LDL TRANSPORT AND PLASMA LDL CONCENTRATIONS ARE REDUCED

This study was undertaken to determine the effect of transient overexpression of hepatic cholesterol 7alpha-hydroxylase on low density lipoprotein (LDL) cholesterol transport in mice lacking LDL receptors (LDL receptor-/-). Primary overexpression of hepatic 7alpha-hydroxylase in LDL receptor-/- mice was accompanied by a dose-dependent decrease in the rate of LDL cholesterol appearance in plasma (whole body LDL cholesterol transport) and a corresponding reduction in circulating LDL cholesterol levels. The increase in hepatic 7alpha-hydroxylase activity necessary to achieve a 50% reduction in plasma LDL cholesterol concentrations was approximately 10-fold. In comparison, cholestyramine increased hepatic 7alpha-hydroxylase activity approximately 3-fold and reduced plasma LDL cholesterol concentrations by 17%. This study demonstrates that augmentation of hepatic 7alpha-hydroxylase expression is an effective strategy for lowering plasma LDL concentrations even in animals with a genetic absence of LDL receptors.

Increased activity of platelet-activating factor acetylhydrolase in low-density lipoprotein subfractions induces enhanced lysophosphatidylcholine production during oxidation in patients with heterozygous familial hypercholesterolaemia.

Patients with heterozygous familial hypercholesterolaemia (FH) have elevated plasma concentrations of low-density lipoprotein (LDL) and develop premature atherosclerosis. There is increasing evidence that oxidative modification of LDL is important for the pathogenesis of atherosclerosis, and the LDL-associated platelet-activating factor acetylhydrolase (PAF-AH) seems to play a key role in LDL oxidation by hydrolysing the oxidized phospholipids of phosphatidylcholine (PC) and producing lysophosphatidylcholine (lyso-PC). We measured the total serum and high-density lipoprotein (HDL) levels of PAF-AH activity and studied the distribution of PAF-AH activity among three LDL subfractions isolated by gradient ultracentrifugation in 15 patients with heterozygous FH and 13 normolipidaemic control subjects. We also determined the lyso-PC production in each LDL subfraction during Cu2(+)-induced oxidation in vitro. The total serum PAF-AH activity in heterozygous FH patients was significantly higher than in control subjects, whereas the HDL-associated PAF-AH activity, expressed as a percentage of total serum PAF-AH activity, was significantly lower in the FH patients than in control subjects (13.9 +/- 6.6% vs. 30.6 +/- 4.4%, P < 0.001). Among the LDL subfractions, the PAF-AH activity in both normolipidaemic control subjects and FH patients, expressed as nmol mg-1 protein min-1, was significantly higher in the LDL3 subfraction (33.1 +/- 4.8 and 53.4 +/- 11.5 respectively) than in the LDL2 (18.6 +/- 5.3 and 26.8 +/- 10.4 respectively, P < 0.0001 for both comparisons) and LDL1 subfractions (5.1 +/- 1.5 and 7.8 +/- 2.6, respectively, P < 0.0001 for both comparisons). Additionally, the enzyme activity in each LDL subfraction of the heterozygous FH patients was significantly higher than in control subjects (P < 0.02 for LDL1, P < 0.03 for LDL2 and P < 0.0001 for LDL3). No difference was observed in the susceptibility to oxidation of each LDL subfraction among the heterozygous FH patients and the normolipidaemic control subjects. During oxidation, the PAF-AH activity decreased, whereas the lyso-PC levels significantly increased in all subfractions of both groups. The lyso-PC/sphingomyelin molar ratio in each LDL subfraction of the FH patients 3 h after the onset of the oxidation was significantly higher than in control subjects [0.38 +/- 0.05 and 0.27 +/- 0.04, respectively, for LDL1 (P < 0.006), 0.47 +/- 0.08 and 0.39 +/- 0.03, respectively, for LDL2 (P < 0.04), 0.55 +/- 0.11 and 0.42 +/- 0.06, respectively, for LDL3 (P < 0.02)]. Our results show that heterozygous FH patients exhibit higher PAF-AH activity than control subjects in all LDL subfractions, resulting in higher lyso-PC production during oxidation, which confers on these subfractions higher biological potency. This phenomenon, in combination with the diminished anti-atherogenic and antioxidant capability of HDL in these patients due to the relatively low HDL-cholesterol levels compared with LDL-cholesterol levels and, consequently, the relatively low HDL-associated PAF-AH activity, could contribute to the higher atherogenicity and incidence of coronary artery disease observed in FH patients.

Adenoviral delivery of low-density lipoprotein receptors to hyperlipidemic rabbits: Receptor expression modulates high-density lipoproteins

Plasma concentrations of low-density lipoproteins (LDLs) and high-density lipoproteins (HDLs) are inversely related in several dyslipoproteinemias. To elucidate the interactions between these lipoproteins, we used a recombinant adenovirus (hLDLR-rAdV) to express human LDL receptors (hLDLRs) in LDL receptor-deficient rabbits. hLDLR-rAdV administration resulted in hepatocyte expression and a reduction of total, intermediate-density lipoprotein (IDL), and LDL cholesterol. In addition, we found that hLDLR-rAdV treatment induced (1) increased very-low-density lipoprotein (VLDL) cholesterol, (2) increased VLDL, IDL and LDL triglycerides, (3) decreased α- and pre-β-migrating apolipoprotein E (apo E) and decreased pre-β-migrating apo A-I at 2 to 4 days posttreatment, and (4) increased total plasma apo A-I and pre-β-migrating apo A-I beginning 8 to 10 days posttreatment. Virtually all plasma apo A-I was present on α- and pre-β-HDL. Pre-β-HDL particles with size and electrophoretic properties consistent with nascent HDL demonstrated the greatest relative apo A-I enrichment following hLDLR-rAdV treatment. In summary, enhanced expression of hepatocyte LDLRs by hLDLR-rAdV treatment markedly altered apo A-I-containing lipoproteins and IDL and LDL. The use of recombinant viruses to express physiologically relevant genes in intact animals, analogous to transfection of cells in culture, provides a new strategy for the evaluation of effects of specific gene products on metabolic systems in vivo.

Statins do more than just lower cholesterol

Clinical trials have demonstrated that inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (statins) greatly reduce cardiovascular-related morbidity and mortality in patients with and without coronary artery disease. 1 Scandinavian Simvastatin Survival Study GroupRandomised trial of cholesterol lowering in 4444 patients with coronary heart disease: the Scandinavian Simvastatin Survival Study (4S). Lancet. 1994; 344: 1383-1389 Summary PubMed Scopus (11241) Google Scholar , 2 Shepherd J Cobbe SM Ford I et al. Prevention of coronary heart disease with pravastatin in men with hypercholesterolaemia. N Engl J Med. 1995; 333: 1301-1307 Crossref PubMed Scopus (7457) Google Scholar These drugs were designed to inhibit the rate-limiting enzyme of cholesterol synthesis in the liver, thereby decreasing hepatic production of lowdensity lipoprotein (LDL), and upregulating expression of hepatic LDL receptors, thus lowering concentrations of circulating LDL. With lower plasma LDL concentrations, plaque development should be retarded or should regress because of lipid loss, which would render the plaque less occlusive and less likely to disrupt or cause thrombosis. DEPARTMENT OF ERRORStatins do more than just lower cholesterol—In this viewpoint by C J Vaughan and colleagues (Oct 19, p 1079), the last sentence of the 2nd paragraph on page 1080 should have read: “In that study a correlation between cholesterol levels and the development of vasculopathy 1 year after transplantation was not apparent”. Full-Text PDF

High prevalence of small LDL particles in non-insulin-dependent diabetic patients with nephropathy

To determine whether small-sized low density lipoprotein (LDL) is associated with a high incidence of coronary heart disease in diabetic nephropathy, we measured the LDL particle size in non-insulin-dependent diabetes mellitus (NIDDM) patients with various degrees of albuminuria ( n = 95) and age-, weight-matched non-diabetic control subjects ( n = 31). The diabetic subjects were divided into three groups, normoalbuminuric, microalbuminuric and macroalbuminuric NIDDM, based on the amount of albuminuria. The average diameter of LDL particles was determined by non-denaturing polyacrylamide gradient (2–16%) gel electrophoresis. The plasma lipid and lipoprotein concentrations were comparable between the non-diabetic controls and normoalbuminuric NIDDM, whereas the plasma triglyceride, very-low-density lipoprotein (VLDL) or LDL concentration was significantly increased in diabetic nephropathy. The mean LDL particle size was significantly smaller in microalbuminuric NIDDM compared with the controls or normoalbuminuric NIDDM, and the LDL size was further decreased in macroalbuminuric NIDDM. The incidence of small LDL (diameter < 255 Å) was remarkably increased in microalbuminuric (58%) and macroalbuminuric NIDDM (67%) compared to the control (13%) and normoalbuminuric NIDDM (27%). Corresponding to the decreased LDL size, the cholesterol content of the LDL was significantly depleted in NIDDM with nephropathy. The high prevalence of small LDL in diabetic nephropathy was also observed even when hypertriglyceridemic or hypertensive subjects were excluded from each group. The increment in triglyceride-rich lipoprotein ( d < 1.006) after oral fat-loading was increased, and postheparin lipoprotein lipase activity was decreased significantly in diabetic nephropathy. These abnormalities were significantly associated with LDL particle size. Multivariate regression analysis revealed that the amount of albuminuria was closely associated with the average LDL particle size, and this association was independent of the plasma triglyceride level. Neither insulin resistance nor glycemic control was directly associated with LDL particle diameter. The present study indicates that LDL particles become smaller in diabetic nephropathy, and this may be associated primarily with abnormal triglyceride metabolism. However, in addition to hypertriglyceridemia, other metabolic abnormalities caused by diabetic nephropathy may also be involved in the pathogenesis of small LDL particles.

Low-density lipoprotein oxidation, antioxidants, and atherosclerosis: a clinical biochemistry perspective

Cardiovascular disease is the leading cause of mortality in westernized populations. An increased concentration of plasma low-density lipoprotein (LDL) cholesterol constitutes a major risk factor for atherosclerosis. Several lines of evidence support a role for oxidatively modified LDL in atherosclerosis and for its in vivo existence. Antioxidants have been shown to decrease atherosclerotic lesion formation in animal models and decrease LDL oxidation; the evaluation of LDL oxidation in vivo is therefore very important. However, there is a paucity of methods for direct measurement of LDL oxidation. Of the direct methods currently available, the preferred ones seem to be the measurement of F2-isoprostanes, autoantibodies to epitopes on oxidized LDL, and the assessment of antioxidant status. Of the indirect measures, the most uniformly accepted procedure is examining the oxidative susceptibility of isolated LDL by monitoring conjugated diene formation.

Effect of beta-carotene supplementation on the concentrations and distribution of carotenoids, vitamin E, vitamin A, and cholesterol in plasma lipoprotein and non-lipoprotein fractions in healthy older women.

We studied the effect of beta-carotene supplementation on the concentrations and distribution in plasma lipoprotein and non-lipoprotein fractions of carotenoids, alpha-tocopherol, retinol, and cholesterol. Ten women ingested either 90 mg of beta-carotene or placebo daily for 3 weeks while residing in their homes and eating their usual meals. Carotenoids (beta-carotene, lycopene, lutein/zeaxanthin), retinol, alpha-tocopherol, and cholesterol were measured in plasma lipoprotein and non-lipoprotein fractions before and after treatment. In the beta-carotene-supplemented group, total plasma beta-carotene increased 14-fold from 0.48 +/- 0.13 to 6.83 +/- 2.12 mumol/L (p = 0.04). Although the greatest increase in beta-carotene was in low-density-lipoproteins (LDL), the magnitude of increase was similar in LDL, high-density-lipoproteins (HDL), and very-low-density-lipoproteins (VLDL). Thus, the relative distribution of beta-carotene in lipoproteins was unchanged: approximately 71% was in LDL, approximately 15% in HDL and approximately 12% in VLDL, before and after beta-carotene supplementation. There were no changes in amounts and distribution in lipoproteins of the other carotenoids, alpha-tocopherol, and cholesterol. There was no change in the amount of retinol in lipoprotein-deficient plasma. There were no changes in total plasma triglycerides. Significant positive correlations were found between LDL- or VLDL-cholesterol and alpha-tocopherol in LDL or VLDL, respectively; between LDL- or VLDL-cholesterol and lutein/zeaxanthin in LDL or VLDL, respectively; and between HDL-cholesterol and beta-carotene in HDL. beta-Carotene supplementation (90 mg/day for 3 weeks) in healthy older women results in an enrichment of all plasma lipoprotein fractions with beta-carotene, but does not alter the relative distribution of beta-carotene in lipoproteins. beta-Carotene supplementation has no effect on the amounts and relative distribution of lycopene, lutein/zeaxanthin, and alpha-tocopherol in lipoproteins, or of retinol in the non-lipoprotein fraction of plasma. Short-term beta-carotene supplementation has no effect on the concentrations of plasma total triglycerides, total cholesterol, HDL-, LDL-, and VLDL-cholesterol.

Dietary polyunsaturated fat modifies low-density lipoproteins and reduces atherosclerosis of nonhuman primates with high and low diet responsiveness

We tested the hypothesis that an increased content of n-6 polyunsaturated fatty acids (principally linoleic acid) in an atherogenic diet of nonhuman primates would decrease atherosclerosis by modifying the composition and decreasing the concentration of plasma low-density lipoprotein (LDL). A species readily susceptible to diet-induced atherosclerosis (cynomolgus monkey) was compared with a less-susceptible species (African green monkey) with dietary cholesterol concentration and saturated or polyunsaturated fat (40% of energy) as variables. In both species, cholesterol concentrations in whole plasma, LDL, and high-density lipoprotein (HDL) were 20-30% lower when polyunsaturated fat was fed, whereas dietary cholesterol increased LDL cholesterol three- to fourfold. LDL was enriched in cholesteryl oleate when saturated fat and cholesterol were fed. Dietary linoleic acid prevented cholesteryl oleate enrichment and promoted cholesteryl linoleate accumulation in LDL. At the same plasma cholesterol concentration, cynomolgus monkeys had higher LDL cholesterol and lower HDL-cholesterol concentrations than did African green monkeys. LDL particle size was significantly (P < 0.001) larger in the group of cynomolgus monkeys fed polyunsaturated fat but tended to be smaller in African green monkeys fed polyunsaturated fat. Dietary polyunsaturated fat protected against coronary artery atherosclerosis in both species. Thus, LDL particle size, per se, was not atherogenic; instead, coronary artery atherosclerosis and cholesteryl oleate enrichment of LDL were more highly correlated. This outcome suggests that information about LDL composition may be more important for understanding the pathogenesis of atherosclerosis than previously suspected.

Adenovirus-mediated transfer of a gene encoding cholesterol 7 alpha-hydroxylase into hamsters increases hepatic enzyme activity and reduces plasma total and low density lipoprotein cholesterol.

Clinical interventions that accelerate conversion of cholesterol to bile acids reduce circulating low density lipoprotein (LDL) cholesterol concentrations. The initial and rate-limiting step in the bile acid biosynthetic pathway is catalyzed by hepatic cholesterol 7 alpha-hydroxylase. To examine the effects of transient primary overexpression of this enzyme on sterol metabolism and lipoprotein transport, we constructed a recombinant adenovirus in which a cDNA encoding rat 7 alpha-hydroxylase is expressed from the human cytomegalovirus immediate-early promoter (AdCMV7 alpha). Syrian hamsters administered AdCMV7 alpha intravenously accumulated transgene-specific mRNA in the liver and demonstrated a dose-dependent increase in hepatic microsomal 7 alpha-hydroxylase activity. The increased conversion of cholesterol to bile acids resulted in a compensatory increase in hepatic cholesterol synthesis. In addition, overexpression of 7 alpha-hydroxylase reduced the rate of LDL cholesterol entry into the plasma space and, in animals maintained on a Western-type diet, restored hepatic LDL receptor expression. As a consequence, plasma LDL concentrations fell by approximately 60% in animals maintained on control diet and by approximately 75% in animals consuming a Western-type diet. Plasma high density lipoprotein cholesterol levels were reduced to a lesser degree. These results demonstrate that transient upregulation of bile acid synthesis by direct transfer of a 7 alpha-hydroxylase gene favorably alters circulating lipoprotein profiles and suggest one potential molecular target for genetic strategies aimed at reducing cardiovascular risk.

Carbohydrate-fat exchange and regulation of hepatic cholesterol and plasma lipoprotein metabolism in the guinea pig

Adult female guinea pigs were fed semipurified diets containing increasing concentrations of saturated fat (2.5%, 7.5%, 15%, and 25% wt/wt) to determine effects of exchanging fat-carbohydrate calories on lipoprotein metabolism. Plasma very—low-density lipoprotein (VLDL) and high-density lipoprotein (HDL) did not vary but plasma low-density lipoprotein (LDL) concentrations increased with increasing fat calories. LDL cholesterol values were 42 ± 25, 61 ± 17, 92 ± 25, and 98 ± 21 mg / dL (mean ± SD, n = 5), respectively. The relative proportion of cholesteryl ester increased and triacylglycerol (TAG) decreased for VLDL, LDL, and HDL as dietary fat increased. Plasma lecithin cholesterol acyltransferase (LCAT) activity was positively correlated with HDL cholesteryl ester content. Hepatic cholesterol and TAG concentrations were highest in animals fed 25% fat ( P < .01). Hepatic apolipoprotein (apo) B E receptor maximal binding capacity (Bmax) was 30% higher in animals fed 2.5% and 7.5% fat as compared with those fed 15% and 25% fat ( P < .01) and inversely correlated with plasma LDL ( r = −.85, P < .01). In contrast, HDL binding to guinea pig hepatic membranes exhibited a significant positive correlation with dietary fat quantity ( r = .98, P < .001), consistent with a dose-response with increasing fat calories. The activity of hepatic 3-hydroxy-3-methyl glutaryl coenzyme A (HMG CoA) reductase was not affected by the amount of dietary fat, whereas the activity of acyl CoA:cholesterol acyltransferase (ACAT) was significantly increased in animals fed 25% fat ( P < .05). Hepatic free-cholesterol and ACAT activity exhibited a positive correlation for all dietary groups ( r = .75, P < .001). These results demonstrate that exchange of saturated dietary fat for carbohydrate calories results in significant modifications in the regulation of metabolic pathways that determine plasma LDL concentrations and hepatic cholesterol homeostasis.

Carbohydrate type and amount alter intravascular processing and catabolism of plasma lipoproteins in guinea pigs.

To test the effects of exchanging dietary complex and simple carbohydrate for fat calories on lipoprotein metabolism, guinea pigs were fed two different fat/carbohydrate ratios: 2.5:58% (w/w) or 25:29% (w/w) with either sucrose or starch as the carbohydrate source. Animals fed high-fat had higher plasma low-density lipoprotein (LDL) and hepatic cholesterol concentrations than animals fed low-fat diets (P < 0.01). The cholesteryl ester content per particle was higher, and the number of triacylglycerol (TAG) molecules was lower in very low density lipoprotein (VLDL) and LDL from animals fed high-fat diets. Intake of high-fat/sucrose resulted in higher plasma LDL concentrations than intake of high-fat/starch, and animals fed low-fat/starch had the highest plasma TAG concentrations associated with VLDL particles containing more TAG molecules, as well as a TAG-enriched LDL. The activity of plasma lecithin cholesteryl:acyl transferase (LCAT) was highest in animals fed high-fat/sucrose, and heart lipoprotein lipase (LPL) activity was higher in animals fed high-fat diets. Hepatic apoprotein B/E (apo B/E) receptor number (Bmax) was increased 21% with low-fat diets (P < 0.01). These results suggest that the hypercholesterolemia induced by high-fat and by sucrose intake are associated with a higher plasma LCAT activity which results in a cholesteryl ester-enriched VLDL which, by the action of LPL, might be more readily converted to LDL through the delipidation cascade leading to downregulation of hepatic apo B/E receptors. The hypertriglyceridemia associated with low-fat intake may result from increased production of VLDL TAG, which would explain the increased TAG content and the higher TAG/CE ratio of VLDL from animals fed the low-fat/starch diet.

Regulation of apolipoprotein B secretion in hepatocytes from Watanabe heritable hyperlipidemic rabbit, an animal model for familial hypercholesterolemia

Regulation of apolipoprotein B-100 (apo B) secretion in the liver in familial hypercholesterolemia (FH) remains largely unknown. In a previous study, we developed a rabbit hepatocyte culture system and investigated a cellular mechanism which regulates apo B secretion from hepatocytes in response to cellular cholesteryl ester contents [18]. Using this system, in the present study, we investigated regulation of apo B secretion in hepatocytes from the Watanabe heritable hyperlipidemic (WHHL) rabbit, an animal model for FH. Incubation with low density lipoproteins (LDL) at concentrations of 50 or 200 μg/ml, which increased both cellular cholesteryl ester and apo B secretion significantly in normal rabbit hepatocytes, did not cause such increases in WHHL rabbit hepatocytes. However, when WHHL rabbit hepatocytes were incubated with LDL at a concentration of 500 μg/ml, a significant increase in cellular cholesteryl ester and apo B secretion was observed. The effect of the increase in cellular level of cholesteryl ester upon apo B secretion in WHHL rabbit hepatocytes was compatible with that in normal rabbit hepatocytes. Indeed, when WHHL rabbit hepatocytes were incubated with LDL at 1680 μg/ml, a concentration comparable to that of WHHL rabbit plasma, the amount of LDL degradation, cellular cholesteryl ester level, and level of apo B secretion were the same as those in normal rabbit hepatocytes that were incubated with LDL at 160 μg/ml, a concentration comparable to that of normal rabbit plasma. In summary, our present study suggests that, at a steady state with such a high plasma concentration of LDL, the hepatic cholesterol content in WHHL rabbits could be set at the same level as in normal rabbits. It was also shown that cellular cholesteryl ester contents had the same regulatory effect on apo B secretion in WHHL rabbit hepatocytes as in normal rabbit hepatocytes. Therefore, we conclude that in the presence of the genetic defect of the LDL receptor, plasma cholesterol in WHHL rabbit is maintained at a concentration such that apo B secretion is similar to that in normal rabbit.

Regulation of Hepatic 7α-Hydroxylase Expression and Response to Dietary Cholesterol in the Rat and Hamster

Although dietary cholesterol raises plasma total and low density lipoprotein (LDL) cholesterol concentrations, the response to a given intake of cholesterol varies enormously among different species and even among individuals of the same species. The mechanisms responsible for differing sensitivity to dietary cholesterol were examined by comparing the rat, which is able to adapt to large fluctuations in sterol intake or loss with little change in plasma LDL levels, with the hamster, where changes in sterol balance strongly influence plasma LDL concentrations. When fed the same cholesterol-free diet, hepatic 7 alpha-hydroxylase activity was 16-fold higher in the rat than in the hamster. As a consequence, rates of hepatic cholesterol synthesis were 20-fold higher in the rat than in the hamster. In both species, hepatic cholesterol synthesis was suppressed > 90% in response to increasing loads of dietary cholesterol. However, the quantitative importance of this adaptive mechanism was much greater in the rat since the absolute reduction in hepatic cholesterol synthesis in the rat (2,110 nmol/h/g) was much larger than in the hamster (103 nmol/h/g). In the rat, the high basal level of 7 alpha-hydroxylase expression was further induced by substrate (cholesterol) allowing these animals to convert excess dietary cholesterol to bile acids efficiently. In contrast, the low basal level of enzyme expression in the hamster was not induced by dietary cholesterol. Thus, the low basal rates of bile acid and cholesterol synthesis coupled with a lack of 7 alpha-hydroxylase induction by cholesterol render the hamster much more sensitive than the rat to the cholesterolemic effects of dietary cholesterol.

Nutritional, Metabolic, and Genetic Determinants of the Progression of Coronary Heart Disease

The St. Thomas' Atherosclerosis Regression Study (STARS) showed that treatment of mild hypercholesterolemia with diet or diet plus cholestyramine favorably influences the course of coronary heart disease (CHD) over 3 years in middle-aged men. The study employed quantitative angiography to measure change in coronary luminal dimensions. Angiographic benefit was paralleled by improvement in clinical outcomes. The study allowed a detailed analysis of the nutritional, metabolic, and genetic determinants of the progression of CHD. Significant associations were found between changes in both focal and diffuse estimates of coronary atherosclerosis and the plasma concentrations of low-density lipoprotein (LDL) cholesterol and apolipoprotein (apo) B, dietary intake at total and saturated fat, and polymorphisms of the apo-AI gene promoter region and angiotensin-converting enzyme (ACE) gene. Up to 38% of the variance in the progression of CHD could be explained by the independent effects of LDL cholesterol and dietary intake of saturated fat. The ACE genotype was only an independent predictor of the progression of CHD in patients whose LDL cholesterol had been substantially reduced with diet and cholestyramine. The clinical significance of the nutritional, metabolic and genetic findings is discussed.

Guar gum effects on plasma low-density lipoprotein and hepatic cholesterol metabolism in guinea pigs fed low- and high-cholesterol diets: a dose-response study

Guinea pigs were fed semipurified diets containing either 0% or 12.5% guar gum (GG) with 0.04% cholesterol or increasing concentrations of GG (0%, 2.5%, 5%, 7.5%, 10%, and 12.5%) with 0.25% cholesterol (by wt). Compared to the 0% GG diet with 0.04% cholesterol, intake of the 12.5% GG diet with 0.04% cholesterol lowered plasma low-density-lipoprotein (LDL) concentrations, the ratio of LDL cholesteryl ester to protein, hepatic cholesterol concentrations, and the activity of acyl-CoA:cholesterol acyltransferase (ACAT), and increased 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase activity and hepatic apo B/E receptor number (P < 0.01). Intake of GG by animals fed 0.25% cholesterol diets resulted in modest effects on hepatic cholesterol pools and plasma LDL concentrations; however, significant negative correlations were found between both plasma LDL cholesterol and hepatic free cholesterol concentrations with the amount of dietary GG (P < 0.05). Hepatic HMG-CoA reductase was suppressed by the 0.25% cholesterol intake, and GG did not reverse this suppression. In contrast, ACAT activity was negatively correlated with the amount of dietary GG (P < 0.05), and GG intake increased the number of hepatic apo B/E receptors at all intakes with the 0.25% cholesterol diets. These results demonstrate that intake of GG significantly alters endogenous cholesterol metabolism by decreasing hepatic cholesterol pools, altering hepatic cholesterol homeostasis, and reducing plasma LDL concentrations.

Effects of exercise training on the chemical composition of plasma LDL.

The purpose of the present study was to determine the effects of exercise training on the chemical composition of plasma low-density lipoprotein (LDL). Thirteen men (mean age +/- SE, 47.2 +/- 1.5 years) were examined before and after 14 weeks of endurance-oriented exercise training (3 to 4 d/wk, 30 to 45 min/d). Although calculated plasma LDL concentrations remained unaltered (3.49 +/- 0.24 versus 3.65 +/- 0.23 mmol/L), changes in the chemical composition of LDL (increased LDL free cholesterol, cholesterol ester, and phospholipid content) were associated with a reduction in adiposity, umbilical girth, and basal plasma insulin and glucose concentration with training intervention. Increases in LDL molecular weight and particle diameter were associated with a reduction in fat mass, plasma triglyceride concentration, and basal plasma glucose concentration with physical activity. The LDL lipid-to-protein ratio also increased (P < .01) with training by 7%, primarily due to an increase in LDL free cholesterol content (P < .01). These findings indicate the formation of LDL particles that are more cholesterol enriched and protein poor with exercise training, which provides additional evidence for the cardioprotective effect of long-term physical activity.

Platelet function in patients with hypercholesterolaemia

Platelets and plasma lipoproteins, particularly low density lipoprotein, have important roles in atherogenesis. Evidence from several sources suggests that important interactions occur between these individual components of the atherogeneic process. Here we review work from our own laboratory on platelet function in normal individuals and patients heterozygous for familial hypercholesterolaemia (FH). Data is presented on the role of platelet noradrenaline and also on altered cellular signalling in platelets from FH individuals who have plasma low density lipoprotein concentrations which are approximately double those seen in normal subjects.

Regulatory effects of individual n-6 and n-3 polyunsaturated fatty acids on LDL transport in the rat

Dietary triglycerides containing predominantly poly-unsaturated fatty acids (PUFAs) are known to reduce plasma total and low density lipoprotein (LDL) cholesterol concentrations relative to triglycerides containing predominantly saturated fatty acids. However, there is little information regarding the independent effects of individual n-6 and n-3 PUFAs on LDL metabolism. The present studies were therefore undertaken to examine the effects of individual n-6 (linoleic acid) and n-3 (alpha-linolenic, eicosapentaenoic, and docosahexaenoic acid) PUFAs on plasma lipid levels and on the major transport processes that determine plasma LDL concentrations. Rats were fed a semisynthetic cholesterol-free diet supplemented with 4% (by wt) linoleic, alpha-linolenic, eicosapentaenoic, or docosahexaenoic acid for 2 weeks. Dietary eicosapentaenoic and docosahexaenoic acids lowered plasma triglyceride concentrations by 62% and 52%, respectively, and lowered plasma cholesterol concentrations by 54% and 43%, respectively. In contrast, dietary linoleic and alpha-linolenic acids had relatively little effect on plasma triglyceride or cholesterol concentrations. Dietary eicosapentaenoic and docosahexaenoic acids increased hepatic LDL receptor activity by 72% and 58%, respectively, and reduced the rate of LDL cholesterol entry into plasma by 36% and 30%, respectively. As a consequence plasma LDL cholesterol concentrations fell by 60% in animals fed eicosapentaenoic acid and 54% in animals fed docosahexaenoic acid. In contrast, these parameters of LDL metabolism were not significantly altered by dietary linoleic or alpha-linolenic acids. Thus, eicosapentaenoic acid and docosahexaenoic acid (the two major n-3 PUFAs present in fish oil) were equally effective in reducing the rate of LDL formation and stimulating hepatic LDL receptor activity, and were much more active in this regard than their parent compound (alpha-linoleic acid) or linoleic acid.

Dietary fish oil modification of cynomolgus monkey low density lipoproteins results in decreased binding and cholesteryl ester accumulation by cultured fibroblasts

Dietary fish oil (FO) has been reported to increase low density lipoprotein (LDL) receptor function resulting in lower plasma LDL concentrations in the rat (Ventura et al. J. Clin. Invest. 84: 528-537, 1989). The purpose of this study was to determine whether dietary FO, as compared to lard, affected the receptor-mediated uptake of LDL by cultured skin fibroblasts. Plasma LDL was isolated by combined ultracentrifugation and column chromatography from cynomolgus monkeys fed diets enriched in FO or lard and the effect of these two dietary fats on the binding of LDL and esterified cholesterol (EC) accumulation by cultured fibroblasts was determined. There was no difference in total plasma or LDL cholesterol concentrations between diet groups. The monkeys fed FO had significantly smaller LDL which, on average, contained less protein, phospholipid (PL), and free and esterified cholesterol compared to the LDL from monkeys fed the lard diet. FO LDL were less effective than lard LDL in competing for binding, internalization, and degradation of a standard 125I-labeled LDL by fibroblasts (11.0 +/- 2.4 vs. 3.0 +/- 0.8 micrograms LDL protein/ml for 50% displacement of binding, respectively; P = 0.013). FO versus lard LDL also resulted in less accumulation of cellular EC after a 24-h incubation with fibroblasts (7.7 +/- 0.2 vs. 13.0 +/- 0.4 micrograms EC/mg protein, respectively; P = 0.0001). In general, cellular EC accumulation was proportional to LDL particle size and LDL apoE/B molar ratio; however, LDL from the lard group resulted in greater EC accumulation even when LDL particle size and apoE content were nearly equivalent between diet groups. When LDL were isolated from the same animals by sequential ultracentrifugation, the lard LDL apoE was reduced 22% compared to column isolated LDL and this resulted in a 32% decrease in cellular EC accumulation. However, for FO LDL, apoE content was reduced 34% by sequential ultracentrifugation but this only resulted in a 10% decrease in EC accumulation. These results suggested that lard LDL contained more receptor-active apoE than FO LDL. We conclude that isocaloric substitution of fish oil for lard in the diet of cynomolgus monkeys results in LDL particles that bind less avidly to LDL receptors and in less EC accumulation in fibroblasts. The decreased binding of LDL from the FO group appears related to their decreased size and CE content as well as the decreased content of receptor-active apoE relative to the lard group.