Hypocholesterolemic effect of dietary soy protein versus casein in rhesus monkeys (Macaca mulatta).

Ten mature female rhesus monkeys were alternately fed semipurified diets containing casein or soy protein for periods of 13 to 17 wk. Each diet was fed for two periods. When the animals were changed from the commercial diet to the semipurified diet containing soy protein, a significant elevation in serum cholesterol occurred. The serum cholesterol levels gradually increased further, when the soy protein in the diet was replaced by casein. However, when subsequently the casein in the diet was replaced by soy protein, a significant decrease in serum cholesterol levels was observed. Finally, when the animals were changed back to the casein diet, a significant increase in serum cholesterol again occurred. Changes in serum cholesterol due to modulations in the type of protein in the diet were reflected mainly by changes in low-density lipoprotein cholesterol. Thus, the results of this study clearly show that the type of protein in the diet markedly affects serum cholesterol levels not only in experimental animals such as the rabbit, as is well known, but also in the rhesus monkey, which is more akin to man.

Comparison of the metabolism of [1,2,6,7-3H(N)]cholesteryl oleate, cholesteryl [9,10-3H]oleate, and cholesteryl [1-14C]oleate labeled lipoproteins in the rat.

The intravascular metabolism of sterol labeled [1,2,6,7-3H(N)]cholesteryl oleate and acyl labeled cholesteryl [9,10-3H]oleate and cholesteryl [1-14C]oleate was compared in the rat, an animal species without plasma cholesteryl ester transfer activity (CETA). In a first series of studies, the metabolism of sterol labeled [1,2,6,7-3H(N)]cholesteryl oleate and acyl labeled cholesteryl [1-14C]oleate was compared, and the two tracers had identical plasma clearance rates when incorporated into human low density lipoproteins (LDL). The 3H sterol labeled cholesteryl ester (CE), however, had a plasma clearance rate lower than the 14C acyl labeled CE when incorporated into rat alpha- and beta-migrating LDL and human or rat high density lipoproteins (HDL). Unesterified 3H cholesterol reappeared in the plasma whereas the 14C radioactivity in the plasma remained associated with the CE. In a second set of studies, LDL and HDL were radiolabeled with cholesteryl [9,10-3H]oleate and cholesteryl [1-14C]oleate. Large amounts of 3H radioactivity that were dialyzable and not associated with the lipoprotein CE reappeared in the plasma during the kinetic studies. The two tracers had identical plasma disappearance rates when the plasma samples were dialyzed. The results of these studies indicate that the nature of the tracer used to trace lipoprotein CE can affect the estimated kinetic parameters of plasma CE.

Radiolabeled cholesteryl ethers trace LDL cholesteryl esters but not HDL cholesteryl esters in the rat.

The intravascular metabolism of cholesteryl [1-14C]oleoyl ester and [1,2-3H(N)]cholesteryl palmityl ether was compared in the rat, an animal species without plasma cholesteryl ester transfer activity (CETA). The tracers had identical plasma disappearance rates when they were incorporated into human or rat low density lipoproteins (LDL). Fractional catabolic rates (FCR) were 0.081 +/- 0.014 h-1 and 0.080 +/- 0.013 h-1 for human LDL ester and ether and 0.098 +/- 0.007 h-1 and 0.101 +/- 0.007 h-1 for rat LDL ester and ether, respectively. In contrast, the ether had plasma disappearance rates that were 24%-25% lower than the ester when they were incorporated into human or rat high density lipoproteins (HDL). FCR were 0.230 +/- 0.020 and 0.173 +/- 0.030 h-1 for human HDL ester and ether and 0.131 +/- 0.020 h-1 and 0.100 +/- 0.017 h-1 for rat HDL ester and ether respectively. Biological screening of the rat HDL preparations did not affect these differences. The results of these studies indicate that in the absence of plasma CETA, cholesteryl ethers can be used to trace LDL cholesteryl esters but not to trace HDL cholesteryl esters.

Time course of the development of hypercholesterolemia in rabbits fed semipurified diets containing casein or soybean protein.

The time course of the development of hypercholesterolemia was studied in rabbits fed semipurified diets containing casein or soybean protein. When the rabbits were transferred from a commercial diet to semipurified diets, a rapid increase in the concentration of serum cholesterol and phospholipids occurred. After only one day on the semipurified diets, the cholesterol and phospholipid levels increased by about 50% on the diet containing soybean protein and more than doubled on the diet containing casein. Further elevations were observed after one and two weeks on the diets. However after one month, a decrease in the group on the diet containing soybean protein was found. The increases in serum cholesterol and the differences between the soybean protein and casein group were mainly attributable to differences in the LDL fraction and to a minor extent to differences in the HDL fraction. The feeding of semipurified diets resulted in a steep increase in the ratio of cholesterol to protein in all the lipoprotein fractions after only one day. This suggests that lipoprotein particles relatively rich in cholesterol were formed. Marked variations in the density profile of the serum lipoproteins were observed between individual rabbits fed semipurified diets.

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.

The effect of semipurified diets containing different proportions of either casein or soybean protein on the concentration of cholesterol in whole serum, serum lipoproteins and liver in male and female rats.

Male and female lean Zucker strain rats were fed cholesterol-enriched semipurified diets containing 2 levels (20% and 50%, w/w) of either casein or soybean protein for a period of 14 weeks. In the female rats, the feeding of casein diets resulted in significantly higher levels of serum cholesterol than when diets containing soybean protein were fed. In addition, the hypercholesterolemic effect of dietary casein could be enhanced by increasing the proportion of this protein in the diet. Modulations in the proportion of dietary soybean protein did not significantly affect the serum cholesterol levels. In the male rats, however, no such differential effects were observed, indicating a difference between male and female rats in susceptibility to the induction of changes in serum cholesterol levels by dietary means. Upon feeding casein diets, both the male and female rats exhibited a shift of cholesterol from the high density lipoproteins to the lipoproteins with a lower density. This effect was more pronounced in the female than in the male rats. Liver cholesterol concentrations were markedly affected by modulations both in the type and proportion of dietary protein in both sexes. The concentration of cholesterol in the liver of the rats was highest in those fed the 50% casein diet and progressively lower in the animals on diets containing 20% casein, 20% soybean protein and 50% soybean protein.

Prevention of hypercholesterolemia in cholesterol-fed chickens by high-casein and high-soybean protein diets.

Chickens were fed cholesterol-enriched semipurified diets containing two levels (20% and 50%, w/w) of casein or soybean protein for 29 days. The ingestion of the cholesterol-enriched diets containing 20% casein or soybean protein resulted in markedly elevated levels of serum cholesterol, compared with the feeding of a cholesterol-free control diet containing 20% soybean protein. However, this hypercholesterolemic response could be prevented by feeding high-casein and high-soybean protein diets (50%). Similarly, lower levels of serum triglycerides and phospholipids were observed in the chickens fed the 50% protein diets compared with those fed the 20% protein diets. The excess of serum cholesterol and phospholipids in the 20% protein groups was mainly carried in the VLDL and to a lesser extent in the IDL fraction; this was associated with a decrease of cholesterol and phospholipids in the LDL fraction. The amount of cholesterol in the liver paralleled the cholesterol concentrations in the serum; the chickens fed the 20% protein diets exhibited significantly higher concentrations of liver cholesterol than those on the 50% protein diets. Thus, in chickens, the proportion of protein in the diet significantly affects the levels of serum and liver cholesterol, whereas there is practically no differential cholesterolemic effect of casein and soybean protein in the diet.

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.

Dietary corn oil versus olive oil enhances HDL protein turnover and lowers HDL cholesterol levels in hamsters.

We studied the effect of dietary olive and corn oil on high-density lipoprotein (HDL) metabolism in golden Syrian hamsters. The animals were fed a semipurified diet containing 0.1% cholesterol and 40 energy % in the form of either olive or corn oil for a period of nine weeks. Hamsters fed corn oil had significantly lower very-low density and low-density lipoprotein (VLDL+LDL) cholesterol concentrations than those fed olive oil (0.98+/-0.24 vs. 1.40+/-0.34 mmol/l, means+/-S.D., n = 12), as well as significantly lower HDL cholesterol concentrations (3.31+/-0.50 vs. 3.91+/-0.12 mmol/l). The binding capacity of 125I-labelled HDL to liver membranes was 33% higher in the hamsters fed corn oil instead of olive oil (571+/-29 vs. 429+/-24 ng HDL protein/mg membrane protein, P<0.05, n = 4). HDL protein kinetics were studied with 125I-HDL using a constant infusion technique. Both HDL fractional catabolic rate (0.255+/-0. 058 vs. 0.121+/-0.023 /h, P<0.01, n = 5) and transport rate (2.386+/-0. 753 vs. 1.218+/-0.101 mg/h, P<0.01, n = 5) were about 2-fold higher in the hamsters fed corn oil. The rate of plasma cholesterol esterification by lecithin: cholesterol acyltransferase (LCAT) was essentially the same for the two diets. It is concluded that the low HDL level in the hamsters fed corn oil diets is linked with increased HDL binding and degradation in the liver and possibly other tissues. Due to increased HDL protein turnover, the capacity for reverse cholesterol transport is increased in hamsters fed corn oil despite the relative low HDL concentrations

Effects of exercise on plasma high-density lipoprotein cholesteryl ester metabolism in male and female miniature swine.

We studied the effects of exercise on high-density lipoprotein (HDL) cholesteryl ester (CE) metabolism in 6 male and 6 female miniature pigs fed a commercial swine diet supplemented with cholesterol and fat. The diets were fed for a total period of 20 weeks. During the last 12 weeks of the feeding period, the pigs were exercised on a motorized treadmill 5 days per week for 45 min/d at a speed of 9.5 to 10.0 km/h at 0% grade. Homologous HDL preparations were radiolabeled with cholesteryl (1-14C)oleate and intravenously administered to the pigs, followed by blood sampling at the appropriate time points and measurement of radiolabeled HDL CE. This was performed while the animals were sedentary and after the exercise period. Plasma cholesterol increased after the exercise protocol from 7.21 +/- 1.90 to 8.50 +/- 2.81 mmol/L (mean +/- SD, n = 6) in the females and from 8.11 +/- 3.61 to 10.07 +/- 3.61 in the males. HDL CE transport rates in female pigs were significantly lower (23%) after the exercise protocol (118 +/- 14 v 91 +/- 14 micromol/h/L plasma). HDL CE transport rates in the males were also lower (11%) after exercise (90 +/- 20 v 80 +/- 18 micromol/h/L plasma), but this effect was not statistically significant. Further, the residence time or life span of HDL CE was significantly longer after the exercise protocol in both male and female pigs. Thus, the results of this study suggest that exercise reduces the transport rate of HDL CE and prolongs the life span of HDL CE in hypercholesterolemic pigs.

Unidirectional transfer in vivo of high-density lipoprotein cholesteryl esters to lower-density lipoproteins in the pig, an animal species without plasma cholesteryl ester transfer activity.

The metabolism of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesteryl esters (CE) was studied in the pig, an animal species without plasma cholesteryl ester transfer activity (CETA). In the first series of experiments, LDL and HDL from normocholesterolemic pigs were radiolabeled with cholesteryl (1-14C)oleate and intravenously administered to two groups of four normocholesterolemic pigs. Radioactive tracer in LDL remained associated with the LDL fraction, and there was no transfer of LDL-CE to HDL. The transport rate (which represents the production and disposal rate) of LDL-CE in normocholesterolemic pigs was 39 mumol CE/h/L. However, radiolabeled HDL-CE were transferred to LDL (25%), and 36% of the LDL-CE mass was derived from the HDL. The transport rate of HDL-CE was 54 mumol CE/h/L, and the flux of HDL-CE to LDL was 14 mumol CE/h/L. There was no accumulation of radiolabeled HDL-CE in very-low-density lipoprotein (VLDL), which suggests that there was no transfer to VLDL. However, this does not rule out the possibility that either the very low levels of VLDL-CE (< 0.09 mmol/L) or the rapid turnover rate of the VLDL pool might have prevented the accumulation of substantial amounts of tracer in VLDL. Therefore, in a second set of experiments, the kinetics of HDL-CE were studied in high-fat-and high-cholesterol-fed pigs with elevated VLDL-CE concentrations (1.92 mmol/L). Hypercholesterolemia was associated with increased transport rates of LDL-CE (165 mumol/h/L) and HDL-CE (78 mumol/h/L) and with an increased flux of HDL-CE to LDL (78 mumol/h/L).(ABSTRACT TRUNCATED AT 250 WORDS)

Endurance training increased HDL cholesteryl ester metabolism in rats.

We studied the effects of endurance training on the metabolism of high-density lipoprotein (HDL, 1.063 less than density less than 1.15 kg/l) cholesteryl ester and proteins in rats fed a cholesterol-rich (1%) semipurified diet. The HDL were labeled with 131I in the apoproteins and with cholesteryl-[1-14C]oleate in the esters. The HDL were intravenously administered to endurance-trained (n = 10) and cage-sedentary (n = 10) rats. Blood samples were taken over the next 36 h while the rats were conscious and feeding. The trained rats had higher plasma HDL cholesterol (0.72 vs. 0.28 mM) and HDL apoprotein (461 vs. 267 mg/l) concentrations than the sedentary rats. The production or disposal rate of HDL cholesteryl ester was higher in the trained rats (1.36 mumol/h) than in the sedentary rats (0.72 mumol/h), whereas the production or disposal rate of HDL apoproteins was similar in the trained (0.64 mg/h) and sedentary (0.60 mg/h) rats. The residence time of the HDL cholesteryl esters (4.72 +/- 0.22 vs. 3.37 +/- 0.21 h) and HDL apoprotein (7.65 +/- 0.36 vs. 4.55 +/- 0.28 h) was longer for the trained than for the sedentary rats. These data indicate that endurance training resulted in a significant change in the metabolism of HDL cholesteryl esters and apoproteins as well as an increase in their concentrations.

The hypercholesterolemic effect of cafestol in coffee oil in gerbils and rats.

Coffee beans contain the diterpene cafestol, which raises plasma cholesterol concentrations in humans. Daily consumption of 2 g coffee oil, which provides approximately 60 mg cafestol (equivalent to 5.7 mg cafestol/MJ), increases plasma cholesterol concentrations by 28%. We studied the effect of cafestol in coffee oil on gerbils and rats to determine whether the pathways that lead to cafestol-induced hypercholesterolemia in humans are also present in other species. We fed coffee oil from the same batch used in humans to female gerbils and rats. Gerbils were fed a semipurified diet containing 0.5% or 5% (w/w) coffee oil (equivalent to 8.7 and 86.8 mg cafestol/MJ, respectively) in the presence or absence of 0.05% (w/w) cholesterol for a period of 10 weeks. When compared with the gerbils fed no coffee oil, the addition of 0.5% coffee oil to the diets did not affect plasma cholesterol. Plasma cholesterol was significantly higher only when 5% coffee oil was fed, both in the absence (1.01 mmol/L, 33% higher) and presence (1.87 mmol/L, 70% higher) of dietary cholesterol. Liver weight was also significantly higher when 5% coffee oil was fed. Rats were also fed diets containing 0.5% or 5% coffee oil (equivalent to 8.7 and 86.8 mg cafestol/MJ) with and without 0.05% cholesterol for 8 weeks. Feeding 0.5% coffee oil compared with no coffee oil resulted in significantly higher plasma cholesterol levels throughout the study both in the absence (0.46 mmol/L, 27% higher) and presence (0.28 mmol/L, 15% higher) of dietary cholesterol. Diets containing 5% coffee oil appeared to be toxic. Thus, coffee oil diterpenes can result in higher plasma cholesterol in gerbils and rats. The failure to observe these effects in previous studies may be due to doses that were too low.

The effect of exercise on plasma lipids and LDL subclass metabolism in miniature swine.

The effects of exercise on plasma lipids and lipoproteins, high density lipoprotein (HDL) subclass cholesterol levels, and low density lipoprotein (LDL) subclass composition and metabolism were studied in Yucatan miniature swine following 2 yr of training. The exercise protocol produced significant training effects. Post-heparin lipolytic activity was also significantly increased. Although plasma cholesterol and triglycerides did not differ significantly (P = 0.08) between the exercised and control groups, multivariate analysis indicated a strong association between lipoprotein lipase (LPL) and HDL2-C (P less than 0.0001). Although HDL-C levels rose only slightly (P less than 0.09) with exercise, a significant shift was noted in the distribution of cholesterol from the HDL3 to the HDL2 fractions, perhaps mediated by the substantial increase in LPL activity. Exercise had little effect on the chemical composition of the major lipoprotein classes; however, the triglyceride content of the lighter LDL1 subclass was significantly reduced. In the more dense LDL2 subclass, exercise resulted in a significant decrease in triglycerides concomitant with a significant increase in free cholesterol levels. In contrast with the small reductions in fractional catabolic rates (FCR) in either subclass, production rates of the exercised group were reduced, which accounted for the reduction in LDL subclass pool size. These data indicate that exercise produces subtle but significant changes in lipoprotein metabolism that have been previously associated with reduced risk of atherosclerosis.

High density lipoprotein cholesteryl ester metabolism in the pony, an animal species without plasma cholesteryl ester transfer protein activity: transfer of high density lipoprotein cholesteryl esters to lower density lipoproteins and the effect of the amount of fat in the diet.

The metabolism of high density lipoprotein cholesteryl esters (HDL CE) was studied in the pony, an animal species without plasma cholesteryl ester transfer protein (CETP) activity. Studies were done in ponies fed a low- (1.5% fat, w/w) and a high-fat diet (11.5%, w/w fat). The ponies fed the high-fat diet had higher plasma HDL CE concentrations (1.08+/-0.15 vs. 0.84+/-0.11 mmol/l, mean+/-S.D., n=6, P<0.01) and plasma lipoprotein lipase (LPL) activities (14.3+/-4.0 vs. 5.7+/-3.4 micromol free fatty acids (FFA)/ml per h, P<0.05) than those on the low-fat diet. Plasma triacylglycerol (TAG) concentrations were lower on the high-fat diets (0.129+/-0.043 vs. 0.180+/-0.050 mmol/l), but these differences were not statistically significant. There was a negative correlation between the levels of plasma TAG (r=0.598, P<0.05) and VLDL CE (r=0.658, P<0.05) on the one hand and the HDL CE concentrations on the other hand. The transport rates of HDL CE were not significantly different between ponies fed high-fat (0.029+/-0.008 mmol HDL CE/h per l plasma) and those fed low-fat diets (0.024+/-0.004). HDL CE were transferred to low density lipoproteins (LDL) and we calculated that the percentage of LDL CE derived from HDL was 0.69+/-0.13 in the ponies fed the low-fat diet and 0.53+/-0.05 in the ponies fed the high-fat diet (P<0.05). The results of these in vivo studies suggest that in ponies, similarly as reported in rats and pigs, HDL CE can be transferred to LDL despite the absence of plasma CETP activity, and that the magnitude of this transfer is related to the levels of HDL CE as induced by the amount of fat in the diet.

The effect of different proportions of casein in semipurified diets on the concentration of serum cholesterol and the lipoprotein composition in rabbits.

The effect of different proportions of casein in semipurified diets on the concentration of serum cholesterol and the lipoprotein composition was studied in rabbits. Low-casein diets (10% w/w) resulted in serum cholesterol levels and growth rates that were lower than high-casein diets (40%). An intermediate proportion of casein (20%) produced intermediate concentrations of serum cholesterol, but only minor differences in food intake and weight gain, compared with the high-casein group. In the animals with the highest values of total serum cholesterol (the 40% casein group), most of the serum cholesterol was transported in the very low density lipoproteins, whereas with moderate hypercholesterolemia (the 20% casein group), the low density lipoproteins were the main carriers of cholesterol. Elevation in lipoprotein cholesterol was associated in all groups with an increased ratio of cholesterol to protein, suggesting the formation of particles relatively rich in cholesterol. When the rabbits on the diet containing 10% casein were subsequently transferred to the 40% casein diet, a steep increase in the level of serum cholesterol occurred. Conversely, switching the rabbits on the 40% casein diet to the 10% casein diet resulted in a decrease in the level of serum cholesterol.

A procedure to secure a jugular vein catheter system onto the neck of the hamster.

We describe a procedure to secure a jugular vein catheter system at the dorsal nape of the neck in the hamster. An 8-cm piece of silicone tubing is connected with a 2.6 cm L-shaped metal tubing which is embedded in prosthetic material. The prosthetic material is placed underneath the neck skin of the hamster and keeps the metal end of the catheter system in a sturdy, upright position.