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During nerve cell degeneration, cholesterol released from the degrading cells is conserved through the formation of a cholesterol-apolipoprotein (apo) E complex which is subsequently taken up by regenerating nerve cells. The aim of the present project was to identify the physiologically relevant lipoprotein receptor for this lipoprotein complex which has remained elusive. HDL was separated into apo E-rich and apo E-poor subfractions and labelled with [14C]-sucrose. Labelled apo E-rich HDL bound to rat brain membranes in a time- and ligand concentration-dependent manner and was a saturable process. Essentially no binding occurred with [14C]-apo E-poor HDL or with free apo E. Binding was partially inhibited by low density lipoprotein (LDL) and by alpha 2-macroglobulin. These results provide new evidence that native apoE-rich HDL particles resembling those present in the brain bind to rat brain membranes and that the binding may be due, at least in part, to the LDL receptor and to the LDL-receptor related protein. Evidence was also provided for the presence of a receptor which binds [14C]-sucrose human apoE-rich HDL in human brain. Characterisation of the receptor which mediates the uptake of cholesterol from HDL-like complexes by brain cells is important in understanding the role of apoE in the central nervous system and of the alterations which occur in disorders such as Alzheimer's disease.

High density lipoproteins (HDL) are small plasma particles which may be able to pass through the blood-brain barrier. We have therefore studied the fatty acids of HDL in patients with dementia to determine whether the changes are consistent with those previously reported in brain tissue. The HDL phospholipid and the HDL cholesteryl ester both showed reduced concentrations of arachidonic acid (20:4n6) as compared to normal controls. HDL may be a useful plasma fraction for study of lipids in neurodegenerative diseases.

High density lipoprotein (HDL) has been implicated in the process of reverse cholesterol transport,by which surplus cholesterol is removed from peripheral tissues and transported to the liver for excretion. It has been suggested that some subfractions of HDL may have a particular role in this process, though the underlying mechanism remains unclear. The present study was aimed at investigating the role of specific subfractions of HDL in reverse cholesterol transport. The interconversion of HDL subfractions in normal and cholesterol-loaded rabbits was studied in vivo. Rabbit HDL was separated by heparin-Sepharose affinity chromatography into six subfractions (HDL(I)-HDL(VI)), which were progressively enriched with apolipoprotein E (apo E), and varied in diameter and composition. Total HDL and its subfractions were individually labelled with 14C sucrose and injected in the rabbits. When rabbits which were not acutely loaded with [3H]cholesterol were injected with 14C-HDL(I), 70% of the label remained in this fraction while less than 5% was recovered in HDL(VI), containing the largest particles and those most enriched in apo E. No label was detectable in the liver of these animals. In rabbits which had received a prior loading of cholesterol, an average of only 18.3% of the 14C label was present in HDL(I) while approx. 40% of the label was recovered in HDL(VI). On average, 5.1% of the total 14C injected in these rabbits was recovered in the liver. It is concluded that two alternative routes for reverse cholesterol transport may be operative. While a continuous cholesterol-clearance route may be provided by particles of HDL of intermediate size, another route may be operative for clearance of excess cholesterol loaded into peripheral endothelial cells.

This review describes recent advances that have been made in the separation of HDL subfractions by physicochemical and immunological methods and the relationship between the particles obtained by the different procedures. The metabolic interconversions that occur as a result of cholesteryl ester transfer protein (CETP), lecithin:cholesterol acyltransferase (LCAT) and lipase activities and their role in the formation of mature HDL and in reverse cholesterol transport are discussed. Also considered are the alterations that occur in the HDL subpopulation profile in coronary heart disease and possible mechanisms by which HDL may protect against this condition.

The concentrations of high density lipoprotein (HDL) subfractions in 100 healthy male subjects were compared with 100 newly presenting patients with myocardial infarction (MI) within 12 h of the onset of chest pain. A subfraction of HDL enriched in apolipoprotein E (apo E), separated by heparin-Sepharose affinity chromatography, was present in lower concentrations (P < 0.001) in the plasma of the coronary patients than in the control subjects. This finding was confirmed by a lower content (P < 0.02) of apo E, measured by ELISA, in the total HDL fraction isolated from the coronary patients. Gradient gel electrophoresis of the total HDL demonstrated that the coronary patients had a significantly decreased concentration of the large HDL particles, HDL2b, of mean diameter 10.57 nm and a higher concentration of the smaller-sized HDL3, especially HDL3c, of mean diameter 7.62 nm. The coronary patients had a lower concentration of HDL cholesterol than the control subjects, attributable to the HDL2 fraction, with no difference in HDL2a between the two groups. There was no difference in the concentration of plasma cholesterol or triglyceride. The distribution of apo E phenotypes was similar in the two groups. HDL2b produced the highest discriminant power between the two groups, followed by apo E-rich HDL, HDL2 and HDL3c Plasma cholesterol correlated strongly with apo E-rich HDL for control subjects but not for MI survivors. This study demonstrates that the inverse relationship between HDL cholesterol and coronary risk shown in epidemiological studies is attributable to the large, apo E-containing HDL subspecies which under some circumstances are implicated in cholesterol removal by reverse cholesterol transport. This study also suggests that the concentration of the large, apo E-containing HDL may provide a sensitive predictor for subjects at risk of developing coronary heart disease.

1. High density lipoprotein (HDL) of the New Zealand White rabbit was separated by heparin-Sepharose affinity chromatography into six distinct subfractions of different composition and particle size. 2. When human acetyl LDL containing [3H]cholesteryl linoleate was injected intravenously into rabbits to prime the endothelial cells with labelled cholesterol, only 1-2% of the radioactivity remained in the plasma after 2 hr. 3. After 4 hr, 60.1% of the plasma radioactivity was present in HDL and 25% of this was recovered in the largest particles of HDL (fraction VI, mean particle diameter 11.6-11.8 nm). 4. The concentration of these largest particles of HDL, rich in apolipoprotein E, were also relatively increased in acetyl-LDL-treated rabbits when compared to controls (P < 0.01). 5. In control in vitro experiments, 62.2% of the radioactivity recovered in HDL was associated with subfractions IV and V (mean particle diameter 10.2-10.8 nm) while only 5% was present in fraction VI. 6. The results show that large HDL particles enriched with apo E contain a large proportion of cholesterol previously supplied to hepatic and peripheral endothelial cells. 7. This study demonstrated that the rabbit provides a useful animal model for the study of the metabolism of subfractions of HDL in relation to reverse cholesterol transport.

The effect of dietary supplementation with evening primrose oil (containing 70% gammalinolenic acid) on the concentration of plasma lipids and lipoproteins of the New Zealand White rabbit was investigated. No significant changes were observed in the concentrations of plasma cholesterol or triglycerides during the treatment, although an increase in high density lipoprotein (HDL) cholesterol (P < 0.01) was observed at 4 weeks of evening primrose oil intake and 2 weeks after withdrawal. However, when HDL subpopulations were resolved by gradient gel electrophoresis, major alterations were observed in the distribution of HDL subfractions. These included an increase in HDL2b (P < 0.001) and HDL3c (P < 0.001) and the appearance of very large particles of HDL. These findings suggest that supplementation of diets with n-6 fatty acids may be effective in the long-term prevention of atherosclerosis.

1. Plasma high-density lipoprotein (HDL) was separated by heparin-Sepharose affinity chromatography into a non-bound, apolipoprotein E-poor, and a bound, apolipoprotein E-rich, fraction through the binding effect of Mn2+ in the column buffer. 2. The application of a series of elution buffers in which the concentration of Mn2+ was progressively replaced by Mg2+ resulted in the separation of the bound HDL into five subfractions. 3. Each subfraction migrated a different distance on gradient-gel electrophoresis. Three of the subfractions had RF (relative migration compared with BSA) values within the range of HDL2b. One subfraction contained largely HDL2a, with some material in the regions of HDL2b and HDL3a, and one subfraction spanned the RF regions of HDL2a, HDL3a and HDL3b. 4. The number of molecules, per HDL particle, of cholesteryl ester, non-esterified cholesterol and phospholipid increased with particle size, whereas triacylglycerol passed through a maximum and the number of amino acid residues remained approximately the same. 5. Apolipoprotein (apo) A-I was the major apoprotein in all five subfractions, but the latter differed appreciably in their contents of apo A-II and apo E. 6. The major fatty acid component of each subfraction was linoleic acid, with moderate amounts of C16:0 and C18:1 fatty acids and a smaller content of C18:0, C20:4,n-6 and C22:6,n-3, with no significant difference in composition between the subfractions. 7. This paper provides the first description of a method for the isolation of three subfractions of HDL2b together with other subfractions in quantities that are sufficient for further analytical or metabolic studies.

The acute effects of running a 42.2 km marathon race on the concentration and composition of the plasma lipoproteins were studied in 56 men of varying fitness, training experience, age and physical characteristics. There was no change in the mean concentration of total serum cholesterol, but a 10.9% increase (P less than 0.001) in the mean concentrations of high-density lipoprotein cholesterol (HDL-TC), representing an 11.1% increase (P less than 0.001) in the cholesteryl ester (CE) and 9.9% increase (P less than 0.001) in the unesterified cholesterol (UC) moieties of HDL. The ratio of total serum cholesterol to HDL-TC decreased significantly (P less than 0.001) during the exercise. Changes in lipoprotein concentrations during the marathon varied considerably between individual subjects, with a small proportion of subjects exhibiting relatively large increases or decreases in HDL-TC, HDL-CE and HDL-UC. Small sub-populations of runners were identified who showed abnormally large decreases in HDL-UC and abnormally small increases in HDL-CE relative to HDL-UC. A correlation (P less than 0.05) was found between the average weekly mileage of training and the increase in HDL-TC, whilst faster runners (finishing time less than 3 h; n = 13) had a significantly greater (P less than 0.02) increase in HDL-TC than slower runners (greater than 4 h; n = 14).(ABSTRACT TRUNCATED AT 250 WORDS)

The distribution of plasma high density lipoprotein (HDL) subfractions was determined in 2 groups of healthy male subjects with different coronary risk indices (CI) as assessed by the ratio of total plasma cholesterol/HDL-cholesterol. The subjects in the 2 groups were of similar age and fitness (as assessed by VO2max). The higher risk group (CI greater than 4.0) contained a lower relative concentration of a specific HDL subfraction, HDL2b, separated by gradient gel electrophoresis, and a lower level of an apo E-rich HDL fraction, isolated by affinity chromatography, than the lower risk group (CI less than 4.0). The concentration of total HDL-cholesterol was higher in the lower risk group due to a difference in HDL2-cholesterol when separation was achieved by polyanion precipitation, but not when separation was made by ultracentrifugation. These observations suggest that the level of these specific HDL subfractions might, when taken in conjunction with plasma cholesterol concentration, provide a better index of coronary risk than that of total HDL as conventionally employed.

The effect of diet on exercise-induced changes in the plasma concentrations of lipoproteins was examined in six healthy male subjects during walks of 37 km on each of four successive days. With a high-carbohydrate diet (85% of the calories as carbohydrate) there was an increase (P less than .05) in the concentration of very-low-density lipoprotein (VLDL)-cholesterol and VLDL-triglyceride and a decrease (P less than .01) in the concentration of high density lipoprotein (HDL)-cholesterol, due mainly to a decrease in HDL3-cholesterol (P less than .01), and HDL-protein (P less than .001). In contrast, a high-fat diet (75% fat) produced a decrease (P less than .01) in the concentration of VLDL-cholesterol and VLDL-triglyceride with increases (P less than .01) in HDL-protein concentration and in HDL-cholesterol concentrations that arose largely from an increase (P less than .001) in HDL2-cholesterol. Gradient gel electrophoretic analysis showed an increase (P less than .01) in the relative concentration of HDL2b (subspecies of diameter 10.57 nm) with a decrease (P less than .01) in the concentration of HDL2a (9.16 nm) plus HDL3a (8.44 nm) with the high-fat diet, but no significant or consistent change with the high-carbohydrate diet. There was no change in the level of the apolipoprotein E-rich HDL subfraction with either diet. Plasma lecithin:cholesterol acyltransferase activity decreased (P less than .05) with the high-fat diet but not with the high-carbohydrate diet. Thus, diet can strongly influence the changes that occur in plasma lipoprotein concentrations during prolonged low-intensity exercise.