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Biomedical subjects

C A Drevon

Publications and source records attributed to C A Drevon.

At least 145 records · Page 8Linked to original sources
PubMed1983

Activity of acyl-CoA: cholesterol acyltransferase and 3-hydroxy-3-methylglutaryl-CoA reductase in subfractions of hepatic microsomes enriched with cholesterol.

The influence of membrane cholesterol on the activities of acyl-CoA: cholesterol acyltransferase and 3-hydroxy-3-methylglutaryl-CoA reductase was examined in three microsomal subfractions (RNA-rich, RNA-poor, and smooth) that had been enriched with cholesterol by incubation with mixed lipoproteins from hypercholesterolemic rabbit serum. Acyl-CoA: cholesterol acyltransferase activity was significantly stimulated in the three subfractions, particularly in the RNA-rich microsomal component. 3-Hydroxy-3-methylglutaryl-CoA reductase, on the other hand, was suppressed (30%) in only one (RNA-poor) of the three microsomal subfractions, despite a 1.4-fold increase in the concentration of membrane cholesterol. An attempt was made to distinguish between an effect based exclusively on an increase in available cholesterol substrate and an activation of acyl-CoA: cholesterol acyltransferase in RNA-rich microsomes enriched with cholesterol. An experimental design was devised so that substrate cholesterol was provided in the form of heated smooth microsomes and acyl-CoA: cholesterol acyltransferase was provided as a separate preparation in the form of RNA-rich microsomes. Appropriate controls were carried out to test for transfer of cholesteryl ester between the two sets of particles. The results suggested that cholesterol enhanced acyl-CoA: cholesterol acyltransferase activity by serving both as a substrate and as a non-substrate modulator.

Acyltransferases↗
PubMed1983

Monensin inhibits receptor-mediated endocytosis of asialoglycoproteins in rat hepatocytes.

Isolated rat liver parenchymal cells incubated in the presence of monensin exhibited a reduced uptake of 125I-asialofetuin (125I-AF). Binding studies indicated that the effect was due to a rapid reduction in the number of active surface receptors for the asialoglycoprotein. Monensin had no effect on receptor internalization, but apparently interrupted the recycling of receptors back to the cell surface. Monensin also inhibited the degradation of 125I-AF previously bound to the cells; this inhibition was probably not due to a direct effect on intralysosomal proteolysis, as no lysosomal accumulation of undegraded ligand could be demonstrated in subcellular fractionation studies by means of sucrose gradients. It is more likely that monensin inhibits transfer of the labelled ligand from endocytic vesicles to lysosomes, as indicated by the accumulation of radioactivity in the former and by the ability of monensin to prevent the normally observed time-dependent increase in the buoyant density of endocytic vesicles. Whereas the effect of monensin on binding and uptake of asialofetuin was reversible, the effect on asialofetuin degradation could not be reversed.

Animals↗
PubMed1983

Metabolism of asialoglycoproteins in cultured rat hepatocytes: evidence for receptor mediated uptake and degradation which is not feed-back regulated.

1. Cultured rat hepatocytes took up and degraded asialofetuin by a saturable mechanism. 2. The uptake was specific for desialylated fetuin; native fetuin and was taken up to a much lesser extent. 3. Degradation was inhibited by lysosomotropic agents; both uptake and degradation were, however, unaffected by cycloheximide. 4. The uptake was also unaffected by incubating the cells for 24 hr in the presence of large extracellular concentrations of unlabelled asialofetuin. 5. The data suggest that the asialoglycoprotein receptor is recycled and not subject to down-regulation.

Ammonium Chloride↗
PubMed1983

Plasma clearance, transfer, and distribution of vitamin D3 from intestinal lymph.

The clearance of vitamin D3 from plasma after intravenous injection to rats of intestinal lymph labeled in vivo with radioactive vitamin D3 has been studied. The half-life of the injected vitamin D3 was about 6 min. In double-label experiments the half-life was the same for vitamin D3 as for vitamin A, which is removed from the circulation as chylomicron remnants. Fractionation of the blood plasma according to the density showed that 2 min after the injection of the lymph 30-95% of the radioactive vitamin D3 was recovered in plasma fractions with higher densities than chylomicron remnants (d greater than 1.019 g/ml). The percentage of vitamin D3 in the chylomicron fraction (d less than 1.006 g/ml) of the injected lymph was 66-95%. Taking this into account it could be calculated that the amount of radioactive vitamin D3 transferred from chylomicrons or chylomicron remnants to plasma with d greater than 1.019 g/ml within 2 min varied from 26 to 72%. The transferred radioactivity was recovered in a protein fraction with chromatographic mobilities identical to the binding protein for vitamin D and its metabolites. During the first 13 min after injection the amount of radioactivity recovered in the liver was less than 10%, but from 15 to 55 min the amount increased to about 50% of the injected dose. These findings suggest that the transfer of vitamin D3 from chylomicrons to the alpha-globulin fraction previously observed in vitro is of importance for the in vivo handling of vitamin D3.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Absorption, distribution, and transport of vitamin D3 and 25-hydroxyvitamin D3 in the rat.

The absorption of [14C]vitamin D3 and 25-[3H]hydroxyvitamin D3 into the mesenteric lymph and portal vein after simultaneous instillation in rat duodenum has been studied. The appearance of both isotopes in the mesenteric lymph and portal vein during the first 2 h was negligible. After 2-4 h the isotopes appeared both in the lymph and portal vein. The appearance of 25-[3H]hydroxyvitamin D3 in the mesenteric lymph was faster than that of [14C]vitamin D3. Total recovery in the lymph during 24 h for [14C]vitamin D3 and 25-[3H]hydroxyvitamin D3 was 15.2 and 14.3%, respectively. While 80% of vitamin D3 in mesenteric lymph was recovered in the chylomicrons (d less than 1.006 g/ml), 94% of 25-hydroxyvitamin D3 was recovered in the protein fraction with d greater than 1.21 g/ml. Seventy-one percent of 25-hydroxyvitamin D3 in the protein fraction was associated with the alpha-globulins. Both vitamin D3 and 25-hydroxyvitamin D3 that were recovered in the chylomicrons (d less than 1.006 g/ml) were transferred in vitro to the lymph fraction with d greater than 1.006 g/ml. Less than 12% of 25-hydroxyvitamin D3 was recovered in the liver after intravenous injection of mesenteric lymph labeled with 25-[3H]hydroxyvitamin D3. Previously, under similar conditions 40% of administered vitamin D3 was recovered in the liver.

Animals↗
PubMed1983

Transport of cholesterol.

.ur current model for cholesterol transport is summarized in Figure 10. In this figure we have put together the various steps in cholesterol transport that were described previously in this review. Under normal conditions, cholesterol metabolism and transport are well regulated. If the transport system is overloaded for a long time, however, hypercholesterolemia caused mainly by increased plasma LDL may develop in several species, including humans. Under such circumstances reverse transport of cholesterol may also fail, giving rise to deposits of cholesterol. Tissue macrophages may be responsible for this lipid accumulation, because receptor-mediated (adsorptive) endocytosis of lipoprotein-associated cholesterol in these cells is not under negative-feedback control. The deposits are mainly found in tissues poorly supplied with blood and lymph: the skin, tendons, the cornea, and arteries. Overload of cholesterol transport may be the result of too much fat and cholesterol in the diet, giving rise to cholesterol-rich lipoproteins from the gut and to increased production of liver (formula; see text) VLDL, which in humans ends up as LDL. In many individuals, however, no hypercholesterolemia is seen, even after eating large amounts of a "western" diet for decades; others may develop increased LDL on a relatively "prudent" diet. Obviously many of the factors and mechanisms in cholesterol transport are influenced by genetic factors. Although studies of several inborn errors of lipid metabolism have given information about some mechanisms, the quantitatively more important differences in genetic patterns, which determine whether or not a western diet will result in hyperlipidemia, are not well known. Perhaps studies of different forms of apoB and apoE and of HDL subgroups and hyper-alpha-lipoproteinemia will explain why certain individuals develop hypercholesterolemia and premature atherosclerosis. All the recent information related to cholesterol metabolism and transport gives rise to new questions. There are many problems of interest for future research: What are the metabolic differences between the apoB produced in the liver and that produced in the gut? To what extent is the protein moiety of LDL modified in the plasma of blood and lymph and in interstitial tissue? Are such modifications important to whether LDL uptake goes through the classic LDL pathway or through the macrophage (i.e., scavenger?) pathway? Are some changes in apoB important for liver recognition of LDL?(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1982

Evidence that reverse cholesterol transport occurs in vivo and requires lecithin-cholesterol acyltransferase.

The transport of cholesterol from extrahepatic tissues into plasma (reverse cholesterol transport) and the possible requirement for lecithin:cholesterol acyltransferase was examined in the rat. One hour after removal of the liver plasma cholesterol ester concentrations were significantly increased by 20%, whereas free cholesterol concentrations were unchanged. The lecithin:cholesterol acyltransferase inhibitor, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) was administered to eviscerated rats. It inhibited plasma lecithin:cholesterol acyltransferase activity by 90% which in turn totally prevented the increase in plasma cholesterol ester concentrations. In addition, heat-inactivated plasma from DTNB-treated eviscerated rats was 50% more reactive toward a standard source of lecithin:cholesterol acyltransferase compared to plasma from control or untreated eviscerated rats. These data suggest that in the rat a reactive lecithin:cholesterol acyltransferase substrate is formed extrahepatically. Together with lecithin:cholesterol acyltransferase, this reactive substrate removes cholesterol from peripheral tissues.

Animals↗
PubMed1982

Metabolism of ochratoxin A by primary cultures of rat hepatocytes.

Association of ochratoxin A with cultured rat hepatocytes occurs at 4 degrees C, and the saturation level in the medium is 0.3 mM ochratoxin A, with maximal binding after 60 min. At 37 degrees C the level of cell-associated ochratoxin A increased up to 6 h and remained at 2 nmol of toxin per mg of cell protein for 30 h. With increasing concentrations of ochratoxin A, increasing amounts of the toxin accumulated in the cells; saturation occurred at a concentration of 0.3 mM. Ochratoxin A was metabolized by hepatocytes at 37 degrees. (4R)-4-Hydroxyochratoxin A appeared in the medium at a maximal level (about 30 nmol/mg of cell protein) at an ochratoxin A concentration of 0.25 mM after 48 h of incubation. Small amounts of (4S)-4-hydroxyochratoxin A were detected only after incubation for 22 h or longer.

Animals↗
PubMed1982

26-hydroxylation of 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol by isolated nonparenchymal cells and hepatocytes from rat liver.

The ability of isolated nonparenchymal and parenchymal rat liver cells to metabolize 5beta-cholestane-3alpha,7alpha,12alpha-triol and other bile acid intermediates has been investigated. Incubation of nonparenchymal cells with 5beta-cholestane-3alpha,7alpha,12alpha-triol resulted in the formation of one more polar product identified as 5beta-cholestane-3alpha,7alpha,12alpha,26-tetrol. The formation was linear with time up to 2 hr and with the number of cells, and showed saturation kinetics with respect to substrate concentration. The maximum rate of conversion was 90 pmol/10(6) cells per hr. Incubation of hepatocytes with the triol resulted in the formation of several more polar products. In addition to 5beta-cholestane-3alpha,7alpha,12alpha,26-tetrol, products with retention time on high pressure liquid chromatography identical with 3alpha,7alpha,12alpha-trihydroxy-5beta-cholestan-26-oic acid and cholic acid were observed. The identity of these products was verified by combined gas-liquid chromatography-mass spectrometry. The rate of conversion was linear with time for about 10 min and saturation with respect to substrate concentration was not attained. At identical substrate concentrations, the total rate of conversion (12.5 nmol/10(6) cells per hr) was at least two orders of magnitude higher than with the nonparenchymal cells. Similar differences in rates of conversion were observed with other C(27)-bile acid intermediates. It is concluded that the nonparenchymal cells do not play any significant role in the conversion of bile acid intermediates, either under physiological conditions or under experimental conditions where such steroids have been administered intravenously.-Dueland, S., J. I. Pedersen, C. A. Drevon, and I. Björkhem. 26-Hydroxylation of 5beta-cholestane-3alpha,7alpha,12alpha-triol by isolated nonparenchymal cells and hepatocytes from rat liver.

Animals↗
PubMed1981

The binding of high density lipoproteins to isolated rat hepatocytes.

The association of 125I-labelled rat high density lipoproteins (125-I-HDL) to suspended rat hepatocytes was studied at 4 degrees C. 125 I-HDL associated to isolated hepatocytes by two processes-one of high and one of low affinity. The cell-association of 125I-HDL exhibited saturation kinetics and was inhibited to varying degrees with both rat and human lipoproteins such as VLDL, LDL and HDL but not by lipoprotein deficient serum or asialo-fetuin. The cell-association of 125I-HDL did not require divalent cations and could be reduced by pronase treatment of the cells. The binding site was clearly different from the receptor for LDL in extrahepatic cells since heparin and apolipoprotein E did not compete and cholesterol ester labelled HDL. The number of binding sites for HDL at 4 degrees C was 2.2 X 10(6) per cell and the association constant (Ka) 8.2 X 10(6) (mol/l)-1. Experiments with HDL labelled with [3H] cholesterol by means of lecithin: cholesterol acyltransferase (LCAT, EC 2.3.1.43) in the cholesterol ester moiety suggested that the same mechanism was responsible for the cell-association of HDL prepared this way.

Animals↗
PubMed1981

Metabolism of homologous and heterologous lipoproteins by cultured rat and human skin fibroblasts.

Rat fibroblasts degraded human low density lipoprotein (LDL) very slowly, one-tenth to one-fortieth the rates observed in human fibroblasts. In rat cells, human LDL caused only very small increases in cell cholesterol content and acylCoA:cholesterol acyltransferase (ACAT) activity and caused only small decreases in beta-hydroxy-beta-methylglutaryl CoA (HMG CoA) reductase activity; in human cells, however, human LDL induced very large changes in all three of these parameters, as expected. The binding of human LDL to rat fibroblasts was not reduced by previous incubation with human LDL or with 25-hydroxycholesterol. Thus, in rat fibroblasts there appear to be few, if any, regulated high-affinity receptors that recognize human LDL. Rat LDL fractions (d 1.02-1.05 g/ml), in contrast, were degraded more rapidly than human LDL by rat fibroblasts, caused a significant increase in cell cholesterol content, an increase in ACAT activity, and a significant decrease in HMG CoA reductase activity. Moreover, the degradation of this rat LDL fraction by rat fibroblasts as a function of concentration was biphasic, i.e., there appeared to be a high-affinity component of degradation. Thus, it appears that rat fibroblasts do have a receptor for homologous lipoproteins. However, because both apoprotein B and apoprotein E are present in these rat lipoprotein fractions, the observed effects may relate to recognition of either or both of these apoproteins. The metabolism and metabolic effects of the conventionally defined high density lipoprotein (HDL) fraction of the rat by rat or human fibroblasts resembled those of human LDL in human fibroblasts. It is suggested that rat HDL may, because of its apo E content and higher concentration in rat plasma relative to that of LDL, play an important role in cholesterol homeostasis in vivo.

Animals↗
PubMed1980

Regulation of cholesterol esterification and biosynthesis in monolayer cultures of normal adult rat hepatocytes.

Adult rat parenchymal liver cells were isolated and cultured in monolayers. Cholesterol esterification in the intact cultured cells was determined by measuring incorporation of tritiated oleic acid into cell cholesterol ester. Addition of 10 microgram/ml of 25-hydroxycholesterol to the medium gave a 3- to 6-fold increase in cholesterol esterification, while the incorporation of oleic acid into phospholipids and triglycerides remained unaltered. Similar stimulation of cholesterol esterification by 25-hydroxycholesterol was also found if [14C]mevalonolactone or [3H]cholesterol (the latter presented to the cells in high density lipoproteins) were used as precursors. The stimulatory effect of 25-hydroxycholesterol was maximal after only 15-min incubation and was independent of protein synthesis. After 4 to 6 h of incubation with 25-hydroxycholesterol, its stimulatory effect was reduced significantly, and after 18 h of incubation no stimulation was observed. Thus, the liver cells in some fashion adapt to the continuing presence of 25-hydroxycholesterol. Isolated microsomes prepared from cells previously incubated with 25-hydroxycholesterol showed acyl-CoA:cholesterol acyltransferase activity 3 times that of microsomes from control cells. Incubation of isolated microsomes with 25-hydroxycholesterol increased acyl-CoA:cholesterol acyltransferase activity 2-fold. The net cellular content of ester cholesterol increased after 2 to 6 h incubation of hepatocytes with 25-hydroxycholesterol; there was a net decrease in cellular free cholesterol. Mevalonolactone (10 mM) also stimulated cholesterol esterification and inreased the cellular content of ester cholesterol (3- to 4-fold). The effectiveness of mevalonolactone did not diminish with longer periods of preincubation. Furthermore, the stimulatory effects of 25-hydroxycholesterol and mevalonolactone added together were at least 50 to 100% greater than the effects of either agent alone, suggesting that the mechanisms by which they increase cellular cholesterol esterification are different. Both 25-hydroxycholesterol and mevalonolactone rapidly inhibited microsomal 3-hydroxy-3-methylglutaryl-CoA reductase activity. Pure cholesterol had no effect on cellular cholesterol esterification or on 3-hydroxy-3-methylglutaryl-CoA reductase activity at concentrations up to 10 microgram/ml.

Animals↗
PubMed1980

Dissociation of the lipid-enzyme complex of hormone-sensitive lipase using high density lipoprotein or apolipoprotein A-I.

Hormone-sensitive lipase in homogenates of adipose tissue occurs as a large, lipid-rich complex including several acylhydrolase activities that emerge quantitatively in the void volume on gel filtration chromatography (2% agarose). Incubation with intact human plasma high density lipoprotein or with lipid-free apolipoprotein A-I, however, disrupted the lipid-rich complex almost completely and most of the enzyme activity eluted from a 2% agarose column at about Ve = 2.3 x Vo. This use of the detergent-like properties of apolipoprotein A-I may be of value for dissociation of other lipid-associated or membrane-bound enzymes.

Adipose Tissue↗
PubMed1980

Influence of cholesterol and fat feeding on hepatic lysosomal enzymes in guinea pigs.

The activity of four lysosomal enzymes have been examined in liver cytoplasmic extract from guinea pigs fed three different diets: a) an ordinary diet, low in fat, high in carbohydrates; b) a semisynthetic diet containing 10% cottonseed oil (by weight) without and c) with 1% cholesterol. The cholesterol content in the liver was similar in control-fed and fat-fed animals, while there was a 10-fold increase in cholesterol + fat-fed animals, and most of this cholesterol was present as ester. We observed increased activity of beta-glucuronidase, beta-acetyl-glucosaminidase and cathepsin D during fat cholesterol feeding (diet c) while the activity of acid phosphatase decreased compared to control-fed animals. These findings probably mirror the increased hepatic accumulation of lipids and lipoproteins observed in cholesterol + fat-fed guinea pigs.

Acetylglucosaminidase↗
PubMed1980

Secretion of very low density lipoproteins enriched in cholesteryl esters by cultured rat hepatocytes during simulation of intracellular cholesterol esterification.

Very low density lipoproteins, newly secreted by cultured rat hepatocytes into a serum-free medium, contain some cholesteryl esters although the percentage of total cholesterol in ester form is less than that in plasma very low density lipoproteins. When acyl CoA:cholesterol acyltransferase activity in hepatocytes was stimulated by the addition of 25-hydroxycholesterol (10 microgram/ml) or mevalonolactone (1 mM), the absolute amount of esterified cholesterol secreted in very low density lipoproteins increased significantly, but the amount of free cholesterol decreased or showed no change. Thus the percentage of very low density lipoprotein cholesterol in ester form increased, in some experiments to as much as 50% of the total. These results provide additional evidence that hepatic acyl CoA:cholesterol acyltransferase plays a role in the generation of some of the cholesteryl ester in newly-secreted lipoproteins. They further suggest that changes in the activity of the enzyme can potentially regulate the fraction of cholesterol secreted in esterified form.

Animals↗
PubMed1979

The hydrolysis of cholesterol esters in plasma lipoproteins by hormone-sensitive cholesterol esterase from adipose tissue.

Adipose tissue contains a high level of neutral esterase active against emulsions of cholesteryl oleate. The present studies show that this enzyme can also effectively hydrolyze the cholesterol esters in native rat plasma high density lipoproteins (HDL) and low density lipoproteins (LDL). The hydrolysis of lipoprotein cholesterol esters by a pH 5.2 isoelectric precipitate fraction from the freshly prepared 100,000 X g supernatant of chicken adipose tissue was low, but increased more than 50-fold on activation with cyclic AMP-dependent protein kinase. Rat adipose tissue homogenates were also very active against lipoprotein cholesterol esters, hydrolyzing as much as 60% of the total labeled cholesterol ester in HDL or LDL in 1 h. Activity was optimal at pH 7 and very low at pH 4. No protease activity was detected at pH 7 and, since assays were done in 2 mM EDTA, phospholipase A activity was presumably negligible. The results show that hormone-sensitive cholesterol esterase of adipose tissue has ready access to the neutral lipid core of plasma lipoproteins, either because the enzyme penetrates the polar shell or because the cholesterol ester in the core is exposed, at least intermittently, to allow enzyme-substrate complex formation. Whether or not this enzyme activity plays a role in lipoprotein degradation by adipose tissue remains to be determined.

Adipose Tissue↗
PubMed1979

Influence of cholesterol/fat feeding on cholesterol esterification and morphological structures in intestinal mucosa from guinea pigs.

Guinea pigs were fed a semisynthetic diet containing 10% (by weight) cottonseed oil and 1% cholesterol. In response to cholesterol/fat feeding there was an increase in both the unesterified cholesterol (UC) and cholesteryl ester (CE) of the intestinal mucosal cell. Along with the increased cholesterol levels there was a 4-fold increase in the microsomal acylCoA:cholesterol acyltransferase (ACAT) activity after only two days of cholesterol/fat feeding. After 6 days on the experimental diet the ACAT activity was up to 8-fold the activity of the control, and then remained at this level for up to 20 days. The increased ACAT activity was probably not due to increased substrate concentration alone, since the fractional esterification of cholesterol also increased when the cholesterol/fat containing diet was given. There was also an increase in the triglyceride content of the intestinal mucosal cells from guinea pigs on the experimental diet. The mucosal cells of the cholesterol/fat fed animals accumulated varying amounts of lipid droplets, which were without an enveloping membrane, suggesting that the uptake of lipids from the intestinal lumen was higher than the capacity to synthesize and/or secrete lipoproteins. Simultaneously the size and amount of secondary lysosomes increased. A considerable increase in lipid droplets, lipolysosomes, and residual bodies was observed in the lamina propria macrophages while no crystalline clefts were seen.

Animals↗