Effect of dietary fat saturation on plasma lipoproteins and high density lipoprotein metabolism of the rhesus monkey.

Rhesus monkeys were fed corn or coconut oil-based diets for 3-6 mo to determine effects on the composition of all lipoprotein classes and on the metabolism of high density lipoproteins (HDL). Major findings included the following. Coconut oil feeding increased concentrations of all classes of plasma lipoproteins without altering lipoprotein size, suggesting an increase in particle number. The percentage of saturated fatty acids in the cholesteryl esters (CE) of low density lipoproteins (LDL) and HDL reached 40% with coconut oil feeding. This value probably constitutes a minimum estimate of the CE which were of intracellular rather than intraplasmic origin. The CE in LDL and HDL were nearly identical suggesting virtually complete equilibration by the core lipid transfer reaction. The CE in very low density lipoproteins, in contrast, were significantly more saturated than those in LDL and HDL irrespective of diet. Lower HDL levels on the corn oil diet were associated with higher fractional catabolic rates for both apolipoprotein A-I (0.42 vs. 0.31 d-1) and apolipoprotein A-II (0.45 vs. 0.30 d-1).

Rapid clearance of serum amyloid A from high-density lipoproteins.

The serum amyloid A proteins (SAA) occur in plasma in six polymorphic forms that are associated with the high-density lipoproteins (HDL). We studied two of the SAA proteins, SAA1 and SAA4, which have the same amino- and carboxy-terminal residues but different solution properties and electrophoretic mobilities, to determine whether they are interconverted in plasma in vivo. They were radioiodinated in vitro, incorporated into HDL, and administered to cynomolgus monkeys. Both remained associated with HDL for at least 6 h, had similar plasma die-away curves, and retained their characteristic electrophoretic mobilities, suggesting they are not related as precursor and product. The plasma clearance of the most prevalent SAA species, SAA4, was also simultaneously compared with the human A-I and C-III-2 apolipoproteins. Human apolipoprotein A-I decayed from plasma at a rate comparable to that of monkey HDL proteins. Apolipoprotein C-III-2 was cleared more rapidly and SAA4 at an even greater rate. These findings suggest that SAA are either dissociated from HDL before clearance from plasma or that SAA are contained in an HDL subspecies with metabolic fate different from that of most HDL particles.

Nucleotide sequences of the Macaca fascicularis apolipoprotein C-III and A-IV genes.

The cynomolgus monkey (Macaca fascicularis) apolipoprotein C-III and apolipoprotein A-IV genes have been isolated from a cynomolgus genomic DNA library and completely sequenced. These genes span 3.1 and 2.8 kilobases (kb), respectively. Apolipoprotein C-III gene is interrupted by three intervening sequences of 613, 135 and 1699 bp, respectively. The open reading frame encodes a protein of 99 amino acids which is 87% similar to the human. The cynomolgus mature protein is 79 residues long. Thr-74 is also present what might allow the formation of O-glycosidic linkage observed in the human protein. Apolipoprotein A-IV gene consist of two intervening sequences of 352 and 774 bp, respectively. The open reading frame encodes a protein of 429 amino acids which is 87% similar to the human. The cynomolgus mature protein is 409 residues long, 33 amino acids longer than the human, due to an insertion of 33 residues in its COOH-terminal region. This insertion is mainly composed of glutamine and glutamic acid, which confers cynomolgus apolipoprotein a higher hydrophilicity.

Glucosamine effects in humans: a review of effects on glucose metabolism, side effects, safety considerations and efficacy.

Glucosamine is widely used to relieve symptoms from osteoarthritis. Its safety and effects on glucose metabolism are critically evaluated in this review. The LD50 of oral glucosamine in animals is approximately 8000 mg/kg with no adverse effects at 2700 mg/kg for 12 months. Because altered glucose metabolism can be associated with parenteral administration of large doses of glucosamine in animals and with high concentrations in in vitro studies, we critically evaluated the clinical importance of these effects. Oral administration of large doses of glucosamine in animals has no documented effects on glucose metabolism. In vitro studies demonstrating effects of glucosamine on glucose metabolism have used concentrations that are 100-200 times higher than tissue levels expected with oral glucosamine administration in humans. We reviewed clinical trial data for 3063 human subjects. Fasting plasma glucose values decreased slightly for subjects after oral glucosamine for approximately 66 weeks. There were no adverse effects of oral glucosamine administration on blood, urine or fecal parameters. Side effects were significantly less common with glucosamine than placebo or non-steroidal anti-inflammatory drugs (NSAID). In contrast to NSAID, no serious or fatal side effects have been reported for glucosamine. Our critical evaluation indicates that glucosamine is safe under current conditions of use and does not affect glucose metabolism.

Dietary fat saturation effects on low-density-lipoprotein concentrations and metabolism in various animal models.

Saturated vegetable oils (coconut, palm, and palm kernel oil) and fats (butter and lard) are hypercholesterolemic relative to monounsaturated and polyunsaturated vegetable oils. The increase in plasma low-density-lipoprotein-cholesterol (LDL-C) concentrations associated with consumption of saturated vegetable oils and fats is largely explained by a decrease in hepatic LDL receptor activity and an increase in the LDL-C production rate. Hepatic LDL receptor activity may be regulated by the messenger RNA concentration of the LDL receptor. The decrease in hepatic LDL receptor activity with saturated fat feeding is associated with decreased hepatic sterol O-acyltransferase activity and, therefore, a reduced inert pool of cholesteryl ester. A putative regulatory pool of cholesterol is increased with saturated fat feeding and suppresses LDL receptor activity, possibly through hepatic messenger RNA regulation. For most studies, an independent effect of a vegetable oil or fat could not be ascertained because there was no neutral control and at least two of the test oils or fats were varied. Animal data for the effects of individual fatty acids on plasma LDL-C concentrations and metabolism are sparse. The evidence suggests that caproic acid (6:0), caprylic acid (8:0), and capric acid (10:0) are neutral with respect to their LDL-C-raising properties and their ability to modulate LDL metabolism. Lauric acid (12:0), myristic acid (14:0), and palmitic acid (16:0) are approximately equivalent in their LDL-C-raising potential by reducing hepatic LDL receptor activity and increasing the LDL-C production rate, apparently via modulation of sterol O-acyltransferase activity. Stearic acid (18:0) appears to be neutral in its LDL-C-raising potential and how it affects LDL metabolism.

Separation of the dietary fat and cholesterol influences on plasma lipoproteins of rhesus monkeys.

In order to isolate the effects of the type of dietary fat from those of dietary cholesterol on the circulating pool of cholesterol, plasma lipoproteins were characterized in juvenile rhesus monkeys fed semipurified diets containing 31% of calories as corn oil or coconut oil. Half the diets contained 300 mg of cholesterol per 1000 kcal. The fatty acids of cholesteryl esters and phospholipids varied significantly with dietary fat such that saturated and monounsaturated fatty acids replaced polyunsaturated fatty acids in low-density lipoproteins (LDL) and high density-lipoproteins of rhesus monkeys fed coconut oil. Dietary cholesterol alone induced small but significant increases in the cholesteryl ester: triglyceride ratio in both very low-density lipoproteins and LDL. Whereas neither saturated fat (coconut oil) nor cholesterol alone substantially altered the plasma cholesterol concentration or lipoprotein profile, together these dietary components interacted synergistically to produce a significant elevation in plasma cholesterol. This was due primarily to a significant rise in the cholesteryl ester fraction of LDL, disproportionate to any change in LDL protein concentration. The data are consistent with current hypotheses that relate parameters of LDL turnover and clearance to their macromolecular structure and physicochemical characteristics.

Altered lipoprotein metabolism in spontaneous vitamin E deficiency of owl monkeys.

Certain owl monkeys (AOT) develop spontaneous hemolytic anemia that responds to vitamin E. The anemia is associated with red blood cell lipid peroxidation and altered red blood cell membrane lipid composition. To investigate these changes, plasma lipid and lipoprotein profiles were characterized in anemic, anemia-susceptible, and anemia-resistant AOT. The plasma vitamin E and vitamin A concentrations were assessed as an index of fat absorption and the effect of corn oil supplementation and vitamin E-selenium injection were measured. Anemia-susceptible AOT had depressed plasma levels of vitamin E and A and an altered lipoprotein metabolism characterized by elevated ratios of low/high density lipoprotein cholesterol and free to esterified cholesterol in these lipoproteins. Vitamin E-selenium injection in anemia-susceptible AOT increased the plasma vitamin E, and vitamin E and corn oil supplements reduced the high density lipoprotein free to esterified cholesterol ratio. The data suggest that the AOT suffer from fat malabsorption and that the consequences (including tocopherol deficiency) result in altered cholesterol metabolism.

Immunologic effects of marine- and plant-derived n-3 polyunsaturated fatty acids in nonhuman primates.

The effect of marine- and plant-derived n-3 polyunsaturated fatty acids (PUFAs) on T cell-mediated immune response was studied in cynomolgus monkeys. Animals were first fed a 14-wk baseline diet; 10 animals were then fed diets containing 1.3% or 3.3% of energy as eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) which the other 10 were fed diets containing 3.5% or 5.3% of energy as alpha-linolenic acid (ALA) for two consecutive 14-wk periods. Both diets significantly decreased the percentage of T cells (except 1.3% EPA + DHA), T helper cells (except 1.3% EPA + DHA and 3.5% ALA), and T suppressor cells. Proliferative response of lymphocytes to T cell mitogens significantly increased after the diet containing 3.3% EPA + DHA. Interleukin 2 production significantly increased after the diets containing 1.3% and 3.3% EPA + DHA. No significant changes in mitogenic response or interleukin 2 production were found after ALA diets. Feeding 1.3% or 3.3% EPA + DHA or 5.3% ALA significantly suppressed prostaglandin E2 production in response to T cell mitogens. Plasma tocopherol concentrations were decreased significantly only in monkeys fed ALA diets. We conclude that after adjustment for the tocopherol concentration, marine-derived n-3 PUFAs but not plant-derived n-3 PUFAs increased T cell-mediated mitogenic response and interleukin 2 production. This is most likely due to diet-induced quantitative differences in cellular fatty acid composition and, thus, in prostaglandin E2 production and tocopherol status.

Rice bran oil lowers serum total and low density lipoprotein cholesterol and apo B levels in nonhuman primates.

The hypolipidemic response of rice bran oil (RBO) was investigated in nonhuman primates fed semi-purified diets containing blends of oils which included RBO at 0-35% Kcals as dietary fat. The studies demonstrated the following: (a) the degree of reduction of serum total cholesterol (TC) and low density lipoprotein cholesterol (LDLC) was highly correlated with initial serum cholesterol levels of the monkey on the stabilization diet; (b) the content of rice bran oil in the diet was the predominant factor influencing serum TC, LDLC and apoB causing up to a 40% reduction in LDLC without affecting high density lipoprotein cholesterol (HDLC) when RBO was the sole dietary oil fed; (c) the cholesterol-lowering capabilities of RBO were not explained by its fatty acid composition. These studies suggest that RBO may be an additional vegetable oil which lowers serum cholesterol levels by unique mechanisms which will require further study.

Estimation of HDL cholesteryl ester kinetic parameters in the cebus monkey, an animal species with high plasma cholesteryl ester transfer activity.

We studied the kinetic parameters of high density lipoprotein (HDL) cholesteryl esters in the cebus monkey, an animal species with high plasma cholesteryl ester transfer activity. HDL were radiolabeled with cholesteryl [1-14C]oleate and intravenously administered to 4 cebus monkeys. The calculated fractional catabolic rate (FCR) of the HDL cholesteryl esters was 0.081 +/- 0.002 (mean +/- SD) h-1 and the calculated residence time was 12.3 +/- 0.3 h. The production or disposal rate of plasma HDL cholesteryl esters was 34.3 +/- 4.5 mumol/h. The radiolabeled cholesteryl esters were rapidly transferred from the HDL to the very low and low density lipoproteins (VLDL + LDL) and the amount of tracer in the VLDL + LDL had already reached a maximum at 3.5 +/- 0.7 h after tracer administration. The estimated fraction of VLDL + LDL cholesteryl esters derived from the HDL was 0.77 +/- 0.06. We also used radiolabeled [1,2-3H(N)]cholesteryl palmityl ether to trace HDL cholesteryl esters, but the ether tracer was more slowly cleared from the plasma and less readily transferred between plasma lipoproteins than the ester tracer.

Gender differences in response to a hypercholesterolemic diet in hamsters: effects on plasma lipoprotein cholesterol concentrations and early aortic atherosclerosis.

Gender is a strong predictor of coronary heart disease (CHD) susceptibility and reports indicate that males are more likely to develop CHD compared to age-matched premenopausal females. To test whether similar gender differences exist in hamsters, 16 male and 16 female F1B Golden Syrian hamsters, aged 10 weeks, were fed a hypercholesterolemic nonpurified diet (HCD) containing 10% coconut oil and 0.05% cholesterol for 12 weeks. Plasma lipid and lipoprotein cholesterol concentrations, LDL oxidative susceptibility, LDL tocopherol concentrations, LDL fatty acid composition, LDL particle size, plasma estradiol and testosterone concentrations, and early aortic atherosclerosis were analyzed. Female hamsters had significantly lower plasma total cholesterol and nonhigh-density lipoprotein cholesterol (nonHDL-C) and greater high-density lipoprotein cholesterol (HDL-C) concentrations compared to male hamsters (-15, -33, and 33%; respectively). Female hamsters had significantly greater LDL particle size (4%), LDL 22:6 (21%) fatty acid, and rate of LDL oxidation (34%) compared to male hamsters. Female hamsters had a significantly higher concentration of plasma estradiol (49%) compared to male hamsters. Female hamsters also had significantly less early aortic atherosclerosis compared to male hamsters (-77%). In female hamsters, aortic fatty streak formation was significantly associated with plasma nonHDL-C (r = 0.76, P<0.0007), LDL particle size (r = -0.66, P<0.005), plasma TC (r = 0.68. P<0.004), and lag phase of LDL oxidation (r = 0.84. P<0.02). In male hamsters, aortic fatty streak formation was significantly associated with plasma nonHDL-C (r = 0.52, P<0.04), plasma TC (r = 0.55, P<0.03), plasma TG (r = 0.79, P<0.0003), and LDL 22:6 (r = -0.78, P<0.03) with no association with any measures of LDL oxidation susceptibility. This study demonstrates that female hamsters have an improved plasma lipoprotein cholesterol profile, larger LDL particle size, and less early aortic atherosclerosis compared to male hamsters fed the same HCD.

The ACAT inhibitor, CI-1011 is effective in the prevention and regression of aortic fatty streak area in hamsters.

The hypocholesterolemic and anti-atherogenic properties of sulfamic acid ((2,4,6-tris (1-methylethyl) phenyl) acetyl) 2,6-bis(1-methylethyl) phenyl ester, the ACAT inhibitor, CI-1011, was tested in 120 male F1B hamsters fed a hypercholesterolemic chow-based diet containing 10%, coconut oil and 0.05% cholesterol plus: (i) no drug treatment (HCD); (ii) 3 mg/kg per day (HCD+3): (iii)10 mg/kg per day (HCD+10); (iv) 30 mg/kg per day (HCD+30) of CI-1011; or (v) 500 mg/kg per day of cholestyramine (CSTY). Plasma samples were collected at 8 and 10 weeks for measurement of total cholesterol (TC), very low-density lipoprotein cholesterol (VLDL-C), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglycerides (TG). For the progression studies, animals were euthanized after 10 weeks for aortic fatty streak area and hepatic cholesterol analysis. For the regression study, a cohort of the HCD was treated with 30 mg/kg per day of CI-1011 (regression) for an additional 8 weeks. The HCD+3, HCD+10, HCD+30 and CSTY lowered plasma TC (25, 32, 34 and 32%, respectively), VLDL-C (62, 74, 71 and 75%, respectively), LDL-C (25, 38, 47 and 46%, respectively) and TG (48, 47, 42 and 45%, respectively). All treatments resulted in a significant lowering of aortic fatty streak area (68, 86, 93 and 94%, respectively) and reduction in hepatic cholesteryl esters (57, 65, 67 and 70%, respectively). Regression of aortic fatty streak area was 90% after 8 weeks of HCD+30 treatment. Also during the regression phase, plasma TC, LDL-C and TG were lowered 23, 33 and 47%, respectively, as well as, hepatic cholesteryl esters (76%). Significant correlations between plasma LDL-C concentration and aortic fatty streak area (r=0.62, P < 0.004) in the HCD+10 group, suggest that CI-1101 altered aortic lipid infiltration primarily by its effect on plasma lipids. However the 30 mg/kg per day dose of CI-1011 which additionally reduced aortic fatty streak area by 51% relative to the 10 mg/kg per day dose was only associated with a 14% further decrease in plasma LDL-C. Finally the 10-fold regression of aortic fatty streak area was associated with only a 35% reduction in plasma LDL-C. These exceptions to the lipid-lesion relationship raise the possibility of additional effects of CI-1011, which may occur independent of or in concert with lipoprotein cholesterol lowering. It is concluded that in hypercholesterolemic hamsters, CI-1011 is approximately 50 times more potent than cholestyramine in cholesterol-lowering, reduction and regression of aortic fatty streak area.

Soy lecithin reduces plasma lipoprotein cholesterol and early atherogenesis in hypercholesterolemic monkeys and hamsters: beyond linoleate.

The current study was designed to investigate the hypocholesterolemic and anti-atherogenic properties of soy lecithin beyond its fatty acid content. In experiment 1, 18 cynomolgus monkeys were divided into three groups of six and fed diets which approximated either the average American diet (AAD), the American Heart Association (AHA) Step I diet, or a modified AHA (mAHA) Step I diet containing 3.4% soy lecithin for 8 weeks. Plasma samples were collected from food-deprived monkeys and analyzed for total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), very low- and low-density lipoprotein cholesterol (non-HDL-C), and triglyceride (TG) concentrations. Group comparisons revealed that monkeys fed the mAHA Step 1 diet had significantly lower plasma TC (-46%) and non-HDL-C (-55%) levels compared to the AAD diet, whereas monkeys fed the AHA Step 1 diet had lesser reductions in plasma TC (-21%) and non-HDL-C (-18%) levels. The monkeys fed the mAHA Step I diet had significantly lower plasma TC (-32%) and non-HDL-C (-45%) compared to the monkeys fed the AHA step diet. Also, only the mAHA Step I diet significantly reduced pre-treatment plasma TC and non-HDL-C levels by - 39 and -51% respectively with no significant effect on plasma HDL-C or TG levels. In experiment 2, 45 hamsters were divided into three groups of 15 and fed the following three modified non-purified diets for 8 weeks: a hypercholesterolemic diet (HCD) containing 10%, coconut oil and 0.05%, cholesterol, HCD plus 3.4%, soy lecithin (+SL), or the HCD with added levels of linoleate and choline equivalent to the +SL diet but no lecithin (-SL). Plasma lipids were determined as in experiment 1 and aortas were perfusion-fixed and Oil Red O stained for morphometric analyses of fatty streak area. Relative to the HCD group, the +SL-treated hamsters had significantly lower plasma TC (-58%), non-HDL-C (-73%) and aortic fatty streak area (-90%). Relative to the -SL group, hamsters fed the +SL diet had significantly lower plasma TC (-33%), non-HDL-C (-50%) and significantly reduced aortic fatty streak area (-79%). In conclusion, the first experiment suggests that the cholesterol-lowering efficacy of the AHA Step I diet can be enhanced with the addition of soy lecithin without reducing plasma HDL-C levels. whereas the second experiment suggest that the hypocholesterolemic, and in particular, the anti-atherogenic properties of soy lecithin cannot be attributed solely to its linoleate content.

Effects of dietary fat saturation on plasma lipoprotein(a) and hepatic apolipoprotein(a) mRNA concentrations in cynomolgus monkeys.

Plasma lipoprotein(a) (Lp(a)) concentration is an independent risk factor for the development of premature coronary artery disease. Although the majority of available data indicates that circulating Lp(a) levels are under strict genetic regulation, there is some evidence that this parameter may be subject to dietary modification as well. The effects of dietary fat saturation on plasma Lp(a) levels and hepatic apolipoprotein(a) (apo(a)) mRNA abundance were examined in ten unrelated cynomolgus monkeys which were fed each of three experimental diets enriched in saturated (SAT), monounsaturated (MONO), or polyunsaturated (POLY) fatty acids in a crossover design consisting of three 13-week periods. Each diet contained 30% of calories as fat with 0.1% dietary cholesterol by weight and differed solely by the isocaloric substitution of fatty acids as 60% of total fat calories. The mean Lp(a) level for these animals during the SAT diet (13.4 +/- 2.4 mg/dl) was significantly greater as compared with those observed during the MONO (8.6 +/- 2.2 mg/dl, P < 0.0003) and POLY (9.3 +/- 2.1 mg/dl, P < 0.002) diets, while the difference noted between the MONO and POLY diets was nonsignificant. Hepatic apo(a) mRNA abundance was decreased in these animals during the MONO diet relative to both the SAT and POLY diets, with only the difference between the SAT and MONO diets achieving statistical significance (P < 0.02). Our data demonstrate that the substitution of dietary SATs with either MONOs or POLYs result in significant reductions of Lp(a) levels in these monkeys. However, only the MONO diet significantly decreased hepatic apo(a) mRNA levels relative to the SAT diet, suggesting that dietary MONOs and POLYs may differ in the manner by which they regulate plasma Lp(a) levels.

Effects of doxazosin, an alpha 1-adrenergic inhibitor, on plasma lipid and lipoprotein levels, low density lipoprotein metabolism and cholesterol absorption in cynomolgus monkeys.

The mechanism(s) by which doxazosin, an alpha 1 inhibitor, regulates plasma low density lipoprotein cholesterol (LDL-C) and apolipoprotein B (apo B) levels were investigated in 'normocholesterolemic' (average total cholesterol (TC) of 218 mg/dl) and 'hypercholesterolemic' (average TC of 350 mg/dl) cynomolgus monkeys. Twelve weeks of doxazosin treatment (1 mg/kg per day) significantly reduced plasma TC and LDL-C levels in both groups while high density lipoprotein cholesterol and apolipoprotein A-I concentrations rose. Despite these changes in plasma lipids, LDL and HDL lipid composition was not affected by doxazosin. The reduction in LDL-C and apo B in the doxazosin-treated 'hypercholesterolemic' group was associated with a significant increase in both receptor-dependent and -independent LDL apo B fractional catabolic rates. Similar associations were noted in the 'normocholesterolemic' group. LDL apo B production or transport rate was not affected by doxazosin. Cholesterol absorption was also significantly reduced by doxazosin which may also contribute to lowering plasma LDL-C levels. These studies suggest that doxazosin treatment can produce beneficial changes in the plasma lipid profile over a wide rage of plasma cholesterol levels by up-regulating LDL fractional clearance.

Lovastatin inhibits diet induced atherosclerosis in F1B golden Syrian hamsters.

3-Hydroxy-3-methylglutaryl-CoA (HMG CoA) reductase inhibitors are the drugs most commonly prescribed in the US to lower blood cholesterol. Previous studies have shown their efficacy in reducing plasma low density lipoprotein (LDL) cholesterol. However, little is known about their effects on preventing diet induced atherosclerosis. We have investigated the changes in lipoprotein profiles and extent of atherogenesis in hamsters (F1B strain) consuming an atherogenic diet with and without lovastatin. Thirty-six animals were randomized into 3 groups of 12 animals each, with similar plasma cholesterol levels. One group of animals received a basal chow diet, and the other two groups a basal diet plus 10% (w/w) coconut oil and 0.05% cholesterol. After 2.5 weeks, one of the groups received the latter diet supplemented with lovastatin (25 mg/kg/day). A second study was carried out in which animals received the same diets, but lovastatin was given during the 10 week period at a dose of 12.5 mg/kg/day. At the end of the experimental period, animals were sacrificed and lipoprotein cholesterol, liver enzymes, and aortic foam cell development were determined. Animals fed the high fat diet plus lovastatin had significantly lower levels of non-high density lipoprotein (HDL) cholesterol than those fed the unsupplemented high fat diet. No differences were observed in mean levels of this parameter between animals fed the low fat diet and those receiving lovastatin. The amount of aortic lipid staining was significantly less in the lovastatin and low fat groups when compared to the unsupplemented high fat groups. These results indicate that lovastatin can prevent diet induced aortic lipid deposition in this animal model.

Influence of dietary unsaturated and saturated fat on the plasma lipoproteins of Mongolian gerbils.

Lipid and apoprotein moieties of the plasma lipoproteins of Mongolian gerbils (Meriones unguiculatus) were compared in animals fed semipurified diets containing either coconut oil (COC) or safflower oil (SAF). COC-induced hypercholesterolemia was associated with elevations in very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL). Saturated fat feeding also resulted in th saturation of fatty acids of triglycerides and cholesteryl esters of VLDL and LDL, but had little effect on HDL fatty acids. Lipoprotein composition and size were not influenced by the type of dietary fat, suggesting that the hypercholesterolemia with saturated fat feeding was due to the circulation of a greater number of lipoprotein particles. The apoproteins of gerbil lipoproteins had molecular weights comparable to other animals. The relative amounts of apoproteins, particularly the apoC peptides, increased with dietary fat saturation.

Studies of cholesterol and bile acid metabolism, and early atherogenesis in hamsters fed GT16-239, a novel bile acid sequestrant (BAS).

The purpose of this study was to compare the efficacy of GT16-239, an alkylated, cross-linked poly(allylamine) bile acid sequestrant with cholestyramine on cholesterol and bile acid metabolism, and early aortic atherosclerosis in hypercholesterolemic male F1B Golden Syrian hamsters. In this controlled study, 42 hamsters were divided into six groups and were fed a chow-based hypercholesterolemic diet supplemented with a 10% oil blend (55% coconut/45% corn), 0.1% cholesterol (w/w) (control) and either 0.9 or 1.2% cholestyramine or 0.2, 0.4 or 0.6% GT16-239 for 13 weeks. Laboratory analyses included evaluating plasma lipoprotein cholesterol and triglyceride concentrations, hepatic HMG-CoA reductase and 7 alpha-hydroxylase activities, fecal excretion of bile acids and neutral sterols, hepatic cholesterol concentrations, and early atherosclerosis (aortic fatty streak area). Relative to the control diet, the 0.6% GT16-239 versus the 1.2% cholestyramine significantly inhibited the elevation of plasma lipoprotein total cholesterol (TC) (-69% vs -40%), high density lipoprotein-cholesterol (HDL-C) (-49% vs -30%), and non-HDL-C (-81 vs -48%) concentrations; increased the activities of both HMG-CoA reductase (1492% vs 62%) and 7 alpha-hydroxylase (175% vs 86%); lowered the concentration of hepatic cholesteryl ester (-94% vs -59%); increased fecal cholesterol concentration (+28% vs -10%); and decreased aortic fatty streak area (-100% vs -86%). Unexpected findings of this comparison were increased fecal concentrations of cholic acid (533%) and chenodeoxycholic acid (400%) and the reduction in lithocholic acid (-50%) in the 0.6% GT16-239 compared to the 1.2% cholestyramine group. In summary, GT16-239 had a greater impact on cholesterol metabolism and early atherosclerosis in hypercholesterolemic hamsters than cholestyramine.

Doxazosin and cholestyramine similarly decrease fatty streak formation in the aortic arch of hyperlipidemic hamsters.

The effect of doxazosin, an alpha-1 adrenergic inhibitor, on atherosclerosis was determined in hyperlipidemic hamsters. Control hamsters fed chow plus 0.05% cholesterol and 10% coconut oil were compared to chow baseline animals, and to those receiving either 10 mg/kg/day doxazosin, or 245 mg/kg/day cholestyramine in the atherogenic diet. During 8 weeks of treatment, plasma lipids, mean arterial pressure (MAP) and heart rate (HR) were measured, then the ascending aortic arch was examined en face. Numbers of subendothelial macrophage-foam cells/mm2 and their average size (microns 2) were determined, and Oil red O staining (micrograms ORO/mm2) was quantitated to estimate lipid accumulation. Ultracentrifugation of control plasma demonstrate that low density lipoprotein (LDL) carried most of the cholesterol, and very low density lipoprotein (VLDL) was rich in triglycerides. Compared to controls, doxazosin and cholestyramine similarly decreased plasma total and LDL plus VLDL cholesterols, and total triglycerides on average by 46%, 61% and 45% respectively. High density lipoprotein cholesterol was unchanged. Doxazosin also reduced MAP by 18% without affecting HR. In all hamsters, foam cells and lipid accumulated in a lesion-prone area characterized by elevated endothelial cell density, and a thick intima of basement membrane-like material layered over "pads" of smooth muscle cells. Compared to controls, doxazosin and cholestyramine uniformly reduced the number of foam cells/mm2, foam cell size and ORO staining on average by 66%, 29% and 56%, respectively. We conclude that doxazosin decreases plasma lipids and inhibits the development of the fatty streak to a similar level as cholestyramine treatment.

The effects of doxazosin on platelet aggregation, platelet adhesion and blood coagulation in cynomolgus monkeys.

The effect of doxazosin, a selective alpha-1 adrenergic inhibitor, on hemostasis was investigated in 9 cynomolgus monkeys. During 12 weeks of doxazosin treatment (1 mg/kg per day), serum lipids, lipoprotein cholesterols, blood coagulation, platelet aggregation and template bleeding times were measured and compared with predrug values. In addition, platelet adhesion to cultured human umbilical vein endothelial cells (HUVEC) in the presence or absence of doxazosin was evaluated. Platelet aggregation was also determined in monkeys following chronic oral exposure to aspirin (162 mg/day). Doxazosin administration was associated with significant reductions in serum total cholesterol (TC) (-16%) and low density lipoprotein cholesterol (LDL-C) (-23%), while high density lipoprotein cholesterol (HDL-C) levels increased 66%. Doxazosin did not alter any parameters of blood coagulation measured; however, bleeding times were increased significantly (33%) in doxazosin-treated animals. Although collagen-stimulated platelet aggregation was not influenced by either chronic doxazosin or aspirin treatment, the maximal extent of ADP-stimulated platelet aggregation was significantly reduced (-26% and -18%, respectively) compared with the control monkeys. Platelets from untreated control animals displayed reductions in the extent of ADP-stimulated aggregation of 13% and 23%, respectively, when incubated in vitro with 200 and 300 micrograms/ml of doxazosin. Additionally, the decrease in aggregation response of platelets obtained from doxazosin-treated monkeys was accompanied by a rapid reversal of platelet aggregation. Adhesion to HUVEC by platelets isolated from doxazosin-treated animals was significantly decreased; however, adhesion was not altered when platelets from untreated control animals were incubated with HUVEC in the presence of doxazosin. Thus, the ex vivo and in vitro studies reported in this communication suggest that doxazosin administration to nonhuman primates is associated with beneficial alterations in plasma lipids, platelet aggregation, bleeding times and platelet adhesion to endothelial cells, parameters which are thought to influence risk of cardiovascular disease in both animals and humans.