Fecal steroid excretion in relation to the development of casein-induced hypercholesterolemia in rabbits

Two groups of nine rabbits fed a purified diet containing soy protein were injected intravenously with [4-14C]cholesterol. One group was fed the same diet for 78 d while the other group was fed a diet containing casein. The feces of the rabbits were collected and analyzed for radioactivity present in the neutral steroid and bile acid fractions. The excretion of neutral steroids and bile acids was lowered within 3 d after soy protein was replaced by casein and before the serum cholesterol concentration was increased. However, in the casein-fed rabbits the specific radioactivity of the bile acids was greater, whereas the amount of excreted bile acids was lower than in the soy protein-fed rabbits. After the serum cholesterol level in the casein-fed animals became constant (d 35), a further injection of [4-14C]cholesterol was given to all animals. Kinetic parameters of cholesterol metabolism according to a two-pool model were estimated from the die-away curve of [4-14C]cholesterol in serum. The size of the rapidly exchangeable pool (pool A) was greater in the casein-fed rabbits than in the soy protein-fed rabbits, whereas the size of pool B was similar in the two groups. The partial turnover rate of pool A was lower, as was the partial turnover of the slowly exchangeable pool (pool B), and the production rate of cholesterol was significantly lower on the casein-fed rabbits than in the soy protein-fed rabbits. This study suggests that the reduced excretion of steroids in casein-fed rabbits could be the cause of the hypercholesterolemia and is not the result of it.     

Effect of cholesterol supplementation on plasma and liver cholesteryl ester fatty acids in rats fed casein or soy protein

The effect of supplementation with cholesterol on plasma and liver cholesteryl ester fatty acids was examined in rats fed diets containing different protein sources. Weanling male rats were fed a semi-purified diet containing 20% casein or 20% soy protein for seven weeks and then 1% (by weight) of cholesterol was added for three more weeks. Rats fed soy protein grew consistently slower than those fed casein. The plasma and liver cholesterol contents were not significantly different between the two protein groups in the unsupplemented rats; however, they were significantly increased in cholesterol-fed animals, particularly in casein-fed rats. Analysis of the cholesteryl ester fatty acid composition in plasma and liver demonstrated that cholesterol-feeding significantly increased the levels of saturated fatty acids, monounsaturated fatty acids and linoleic acid in casein-fed rats as compared to those fed soy protein. The level of cholesteryl arachidonate in liver, which was low initially, increased slightly in the liver of casein-fed rats but did not significantly change in the plasma of either protein group.     

Fish protein hydrolysate reduces plasma total cholesterol, increases the proportion of HDL cholesterol, and lowers acyl-CoA:cholesterol acyltransferase activity in liver of Zucker rats

There is growing evidence that soy protein improves the blood lipid profiles of animals and humans. We compared the effects of fish protein hydrolysate (FPH), soy protein, and casein (control) on lipid metabolism in Wistar rats and genetically obese Zucker (fa/fa) rats. In Zucker rats, FPH treatment affected the fatty acid composition in liver, plasma, and triacylglycerol-rich lipoproteins. The mRNA levels of Delta 5 and Delta 6 desaturases were reduced by FPH and soy protein feeding compared with casein feeding. In Zucker rats both FPH and soy protein treatment reduced the plasma cholesterol level. Furthermore, the HDL cholesterol:total cholesterol ratio was greater in these rats and in the Wistar rats fed FPH and soy protein compared with those fed casein. Although fecal total bile acids were greater in soy protein-fed Zucker rats than in casein-fed controls, those fed FPH did not differ from the controls. However, the acyl-CoA:cholesterol acyltransferase activity was reduced in Zucker rats fed FPH and tended to be lower (P = 0.13) in those fed soy protein compared with those fed casein. Low ratios of methionine to glycine and lysine to arginine in the FPH and soy protein diets, compared with the casein diet, may be involved in lowering the plasma cholesterol concentration. Our results indicate that the effects of FPH and soy protein on fatty acid metabolism are similar in many respects, but the hypocholesterolemic effects of FPH and soy protein appear to be due to different mechanisms. FPH may have a role as a cardioprotective nutrient.     

Comparison of the hypocholesterolemic effects of dietary soybean protein with those of formaldehyde-treated casein in rabbits

Treatment of casein with formaldehyde changes its tertiary structure and decreases its hypercholesterolemic properties in rabbits. To investigate whether formaldehyde-treated casein exerts this hypocholesterolemic effect in the same manner as soybean protein, rabbits were fed high or low cholesterol diets containing soybean protein, casein, formaldehyde-treated casein or a mixture of casein and formaldehyde-treated casein. Formaldehyde-treated casein was hypocholesterolemic when fed in a low, but not in a high, cholesterol diet. The hypocholesterolemic effect of soybean protein was independent of the amount of cholesterol included in the diet. In contrast to rabbits fed soybean protein, steroid absorption in those fed formaldehyde-treated casein did not differ from that in rabbits fed native casein. Furthermore, the absorption of phosphorus and nitrogen was lower in rabbits fed formaldehyde-treated casein than in those fed native casein, whereas the absorption found in rabbits fed soybean protein resembled that of their casein-fed counterparts. The diets containing soybean protein and formaldehyde-treated casein produced a comparable ratio of lysine to arginine in serum. The results presented in this paper indicate that the hypocholesterolemic action of dietary formaldehyde-treated casein does not resemble that of soybean protein.     

Effects of dietary proteins on lipogenic enzymes in rat liver

When fasted rats were fed fat-free diets containing various sources of protein for 3 d, the activities of liver glucose-6-phosphate dehydrogenase, malic enzyme, acetyl-CoA carboxylase and fatty acid synthetase were markedly lower in rats fed soybean protein or gluten than in those fed casein or fish protein. Since malic enzyme mRNA activity was not low in the soybean protein-fed animals, the translation of malic enzyme appears to be suppressed by dietary soybean protein. The incorporation of tritiated water into liver fatty acids was significantly lower in animals fed soybean protein than in those fed casein. The triglyceride levels in plasma and especially in liver were also lower in the groups fed soybean and gluten than in the groups fed casein and fish. In addition, when dietary soybean protein was replaced with amino acids to simulate casein or soybean protein, the effects on the levels of lipogenic enzymes were still found but were not as great. Thus, some effects can be ascribed to the protein itself and some to the amino acid composition of the diet.     

Effects of addition of sulfur-containing amino acids and glycine to soybean protein and casein on serum cholesterol levels of rats

The effects of the supplementation of methionine (Met), cystine (Cys), and glycine (Gly) to soybean protein or casein on serum and liver lipid levels were studied in rats. Rats were fed cholesterol-free diets containing 25% soybean protein or casein supplemented with 0.75% Met, 2.5% Gly, or a combination of these two for 4 weeks. The addition of Met to soybean protein caused a significant increase in serum cholesterol and this was slightly ameliorated when Gly was given simultaneously. In rats fed casein diets, serum cholesterol tended to decrease when Gly, or Met and Gly were added. A simultaneous supplementation of Met and Gly to casein resulted in a reduction of hepatic cholesterol. Cystine added at the 0.6% level did not cause demonstrable changes in lipid concentrations except for a drop in serum triglyceride of the casein group. When 2.0% Gly was added to cholesterol-enriched diets containing 20% protein, serum cholesterol decreased significantly only when the protein source was casein and the level attained was comparable to that observed in rats fed soybean protein. Liver cholesterol was also markedly decreased by the addition of Gly to casein. The results suggest a possible role of Gly in the regulation of serum cholesterol levels by dietary protein.     

Inhibitory effect of dietary soybean protein vs. casein on magnesium absorption in rats

The effects of casein and soybean protein on magnesium absorption and magnesium concentration in the femur were investigated in rats. Purified diets containing either casein or soybean protein and three concentrations of added magnesium (0.82, 1.64 or 2.46 mmol/100 g diet) were used. The isonitrogenous diets were carefully balanced for the different mineral concentrations in the protein preparations. Absolute and percent magnesium absorption and urinary magnesium excretion were significantly decreased in rats fed soybean protein when compared with casein, irrespective of the dietary concentration of added magnesium. The magnesium content of femur was significantly lower in rats fed soybean protein, but this effect was seen only when the diet contained 0.82 mmol magnesium/100 g diet. The addition of sodium phytate to the casein diets, to a concentration identical to that in the diets containing soybean protein as provided by the soybean protein preparation, produced similar effects on magnesium absorption as the diets containing soybean protein. These results indicate that soybean protein, when compared with casein, decreases magnesium absorption through its phytate component.     

Biliary steroid excretion in rats fed soybean protein and casein or their amino acid mixtures

Male Wistar rats were fed 1% fat, cholesterol-free semipurified diets containing soybean protein isolate and casein or amino acid mixtures simulating these proteins for 28-30 days. The animals then underwent surgery for biliary diversion, and bile was collected for 2 hours. The rate of bile flow was not influenced by the type of dietary protein. The concentration and output of biliary cholesterol in rats fed soybean protein were significantly higher than in those fed casein, while biliary bile acid excretion was comparable. The group given the soy protein-type amino acid mixture also tended to excrete more biliary cholesterol relative to the casein-type amino acid mixture, although the difference was not significant, while biliary bile acid output was comparable. When 5% fat diets containing 0.5% cholesterol were given to rats, concentrations of both biliary cholesterol and bile acid were elevated significantly on a vegetable protein diet. In these experiments, there was a significant negative correlation between serum cholesterol and biliary cholesterol levels. Stimulation of the biliary excretion of cholesterol seems at least relevant to the cholesterol-lowering action of soybean protein.     

Effects of dietary animal and plant proteins on the cholesterol metabolism in immature and mature rats

Three- and 9-mo-old rats were fed purified diets that contained either casein, cottonseed or soybean protein for 28 d, and plasma total and high density lipoprotein (HDL) cholesterol, lecithin cholesterol acyltransferase (LCAT) activity and excretion of fecal neutral sterols were measured. These analyses were performed in order to examine how various dietary proteins from animal and plant sources fed in a purified diet influence the changes in the cholesterol metabolism of the young and old rats. Both immature (3-mo-old) and mature (9-mo-old) rats fed purified diet containing casein maintained significantly higher plasma total and HDL cholesterol levels than their counterparts fed the same diets but containing plant proteins (soybean and cottonseed). The fractional rate of esterification (FR) of plasma free cholesterol in mature casein-fed rats was lower than that in immature rats. The FR was also lower in immature rats fed casein than in those fed plant protein. The net turnover rate (NR) of plasma cholesteryl esters (CE) tended to be higher in mature rats and in general was not affected by the dietary protein source. The rate of fecal excretion of neutral sterols was significantly higher in immature rats than in mature rats and in animals fed plant proteins at both ages than in those fed casein.     

Excessive cholesterolemic response in analbuminemic rats fed a cholesterol-rich diet containing casein.

Female Nagase analbuminemic rats and Sprague-Dawley rats were fed purified diets with or without 1% cholesterol and containing either soybean protein or casein. After consuming the cholesterol-free diets, the analbuminemic rats had significantly higher plasma cholesterol and triglyceride concentrations than the Sprague-Dawley rats. The higher plasma cholesterol levels were essentially in the low density and high density lipoproteins. Based on the fact that the analbuminemic rats excreted more bile acids in feces it is possible that the higher baseline plasma cholesterol concentrations in Nagase analbuminemic rats were partly caused by overproduction of cholesterol. The Nagase analbuminemic rats displayed a greater cholesterolemic response to cholesterol feeding than Sprague-Dawley rats, but only if casein was the protein source in the diet. Casein vs. soybean protein in either cholesterol-free or high cholesterol diets reduced bile acid excretion in Sprague-Dawley but not in Nagase analbuminemic rats. The increased sensitivity to casein plus cholesterol feeding in Nagase analbuminemic rats may be caused by a lack of inhibition of de novo cholesterol synthesis.     

Consumption of casein instead of soybean protein produces a transient rise in the concentration of sphingomyelin in VLDL in rats

In rats fed cholesterol-rich diets, dietary casein vs. soybean protein raises VLDL cholesterol concentrations. Because sphingomyelin may be an essential, structural component of VLDL, we tested whether casein feeding would raise VLDL-sphingomyelin. Rats were fed cholesterol-rich semipurified diets containing either soybean protein (35 g/100 g) or casein for up to 21 d. Consistent with previous work, casein consumption increased hepatic and VLDL cholesterol concentrations. Dietary casein also significantly raised the amount of sphingomyelin in the VLDL fraction, but this effect was transient. Casein feeding transiently lowered LDL- and HDL-2-sphingomyelin concentrations. We suggest that an increase in hepatic VLDL secretion after casein consumption imposed an increased demand for sphingomyelin in the liver. The activity of key enzymes of sphingomyelin synthesis, i.e., serine palmitoyltransferase, phosphatidylcholine:ceramide phosphocholinetransferase and phosphatidylethanolamine:ceramide phosphoethanolaminetransferase and sphingomyelin degradation, i.e., acid sphingomyelinase, were enhanced and depressed, respectively, by casein consumption. Again these effects were transient. Thus, these data indicate that the extra sphingomyelin needed after short-term casein feeding came about through enhanced rates of biosynthesis and reduced rates of degradation in the liver. In addition, plasma transfer of sphingomyelin from HDL-2 to VLDL might have contributed to the increase in VLDL sphingomyelin in the casein-fed rats. This study shows that dietary casein vs. soybean protein transiently influences sphingomyelin metabolism in rats.     

Effects of feeding oyster,Crassostrea gigas,on serum and liver lipid levels in rats

The effects of feeding dietary and defatted oyster meat on lipid metabolism were investigated in rats by comparing measurements with those of casein and soybean protein. In the first experiment, male rats were fed 0.1% and 1% cholesterol-supplemented diets containing casein, oyster or soybean protein under the same dietary level of protein (20%). The concentrations of serum and liver cholesterol in the oyster group were significantly lower than those in the casein group for both the 0.1% and 1% cholesterol-supplemented diets. The cholesterol-lowering effect of oyster meat was more predominant than that of soybean protein. Feeding oyster meat significantly decreased the serum triglyceride concentration as compared to feeding casein for the 0.1% cholesterol-supplemented diets, and it reduced hepatic triglyceride concentration in both groups fed the 0.1% and 1% cholesterol-supplemented diets. The excretion of fecal total steroids was higher in the rats fed oyster meat than those fed casein or soybean protein for both the 0.1% and 1% cholesterol-supplemented diets. In the second experiment, the effects of defatted oyster on lipid metabolism were compared with casein and soybean protein in diets supplemented with cholesterol. The serum cholesterol concentration in the defatted oyster group was comparable to that in the other two groups, but the ratio of high-density lipoprotein-cholesterol to total cholesterol was higher in the defatted oyster group. The feeding of defatted oyster induced a lower liver cholesterol concentration as compared to casein and soybean protein. Serum and liver triglyceride levels were lower in the defatted oyster group than in the casein group. Defatted oyster accelerated the fecal excretion of both neutral and acidic steroids as compared to casein. Our results suggest that the feeding of oysters exerts a more potent hypolipidemic activity than soybean protein, and the effect may be ascribed to both lipid and non-lipid fractions in oyster.     

Relationship between amino acid composition of diet and plasma cholesterol level in growing rats fed a high cholesterol diet

The effects of dietary sulfur-containing amino acids and glycine on plasma cholesterol level were studied in rats fed amino acid mixture diets containing cholesterol. The relationship between the amino acid composition of dietary proteins and plasma cholesterol levels was also investigated in rats fed diets containing various kinds of protein such as casein, egg albumin, pork protein, fish protein, corn gluten, wheat gluten and soy protein. Feeding the amino acid mixture corresponding to casein led to an approximately two-fold level of plasma total cholesterol as compared with feeding the amino acid mixture corresponding to wheat gluten. It was possible to reduce the plasma cholesterol of rats fed the amino acid mixture of the casein type by increasing the proportion of cystine in the total sulfur amino acids. Inversely, the deprivation of cystine resulted in an enhancement of the plasma cholesterol of rats fed the gluten type amino acid mixture. Glycine had a tendency to resist increases in the plasma cholesterol level. A significant negative correlation was noted between plasma cholesterol levels and the content of cystine in intact dietary proteins. The results suggest that the differential effect of dietary proteins on plasma cholesterol level is mainly associated with sulfur-containing amino acids included in the protein, regardless of whether it is of animal or plant origin.     

The quality and quantity of dietary protein affect brain protein synthesis in rats.

The influence of the amino acid supply in diets with different quality and quantity of protein on the rate of protein synthesis in the brain was investigated. Amino acid concentrations in serum and brain altered in accordance with the amino acid levels of the diets, with the exception of some amino acids such as aspartic acid, glutamic acid, glycine and threonine. When rats were fed various levels of dietary casein (0, 5 and 20%), the aggregation of ribosomes increased and the fractional rate of protein synthesis tended to increase with the increase in dietary protein. When rats were fed a 20% casein diet, greater aggregation of brain ribosomes and protein synthesis rate were observed compared with those in rats fed 20% wheat gluten or gelatin diets. The RNA activity was related to the degree of the aggregation of brain ribosomes and the fractional rate of protein synthesis.     

The importance of the non-protein components of the diet in the plasma cholesterol response of rabbits to casein

To characterize the hypercholesterolaemic effect of casein further, four groups of young male rabbits in two separate experiments were placed on cholesterol-free semi-purified diets for 12 weeks. The diets were similar in composition, with either casein or soya-bean-protein isolate providing the protein source (250 g/kg). In two of these diets the salt mix was reduced by 45% (normally 40 g/kg) and replaced by potassium bicarbonate. Growth was unaffected by these alterations in dietary salts except for one group given the soya-bean-reduced-salts diet. The mean concentrations of plasma cholesterol were significantly higher in all casein-fed groups as compared with their soya-bean-fed counterparts but the response was much greater in those given the casein-reduced-salts diet. Contrary to expectations, analysis of the diets showed the zinc and copper concentrations of the casein diets to be less than those of the soya-bean diets. This was due to the greater concentrations of Cu (threefold) and Zn (twofold) in the soya-bean-protein isolate compared with casein. The mean concentration of Zn in fur was significantly decreased in casein-fed rabbits and these animals also excreted less Zn but more Cu in their urine than those given the casein-reduced-salts diet. The rabbits given the casein diet with the least salt mix showed the greatest degree of hypercholesterolaemia, suggesting an interaction between trace elements and the casein effect.     

大豆蛋白对大鼠血浆胆固醇影响及作用机制

目的探讨膳食大豆蛋白对高胆固醇模型大鼠血浆胆固醇及其相关血脂指标的影响及可能的作用机制。方法4周龄断乳雄性Wistar大鼠，诱导高胆固醇模型28d后，按体重和血浆总胆固醇浓度均衡分为2组，分别喂饲含酪蛋白和大豆蛋白的纯合成高脂饲料56d。结果酪蛋白组和大豆蛋白组大鼠TC、甘油三酯（TG）、载脂蛋白B（apoB）含量显著降低，粪胆汁酸排泄量增高。但对高密度脂蛋白胆固醇（鼢L—C），载脂蛋白AⅠ（apoA—Ⅰ）zk平无影响；同时可见，大豆蛋白可以增强大鼠肝脏3-羟基-3-甲酰基CoA（HMG—CoA）还原酶及低密度脂蛋白受体（LDL—R）基因的表达。结论大豆蛋白可通过影响apoB、HMG—CoA还原酶及LDL—R基因的表达来改变血浆胆固醇水平，其作用机制还需进一步探讨。     

Hypocholesterolaemic effect of water-insoluble fish protein from Alaska pollock in ovariectomised rats is not abolished by methionine addition

The present study investigated whether the hypocholesterolaemic effect of water-insoluble fish protein (IFP) from Alaska pollock in ovariectomised (OVX) rats was affected by methionine (Met) addition. OVX rats (6 months old) were fed a cholesterol-free diet containing casein, IFP or IFP+Met as a protein source for 28?d. The ratio of Met:glycine was lower in the IFP and IFP+Met diets compared with the casein diet. Body-weight gain, food intake and liver lipids were not affected by the diet. Plasma total cholesterol concentration was lower in OVX rats fed the IFP diet compared with those fed the casein diet. The hypocholesterolaemic effect of the IFP diet was not abolished by Met addition. Amount of bile acids in the small-intestinal content and faecal excretion of bile acids were higher in OVX rats fed the IFP and IFP+Met diets compared with those fed the casein diet. Ileal bile acid transporter (IBAT) mRNA level and faecal excretion of bile acids were significantly lower and higher, respectively, in OVX rats fed the IFP diet compared with those fed the casein diet, but not in those fed the IFP+Met diet. Thus, the hypocholesterolaemic effect of the IFP diet seems to be mediated by increased faecal excretion of bile acids coupled with decreased reabsorption of bile acids from the ileum through a decrease in IBAT and the change in cholesterol metabolism linked to the amino acid profile.     

VLDL metabolism in rats is affected by the concentration and source of dietary protein

The present study was designed to determine if changes in dietary protein level and source are related to changes in VLDL lipid concentrations and VLDL binding by hepatic membranes and isolated hepatocytes. Male Wistar rats were fed cholesterol-free diets containing 10, 20 or 30 g/100 g casein or highly purified soybean protein for 4 wk. Hepatic, plasma and VLDL lipids, VLDL apo B-100 and VLDL uptake by isolated hepatocytes and VLDL binding to hepatic membrane were determined. Increasing casein or soybean protein level (from 10 to 30 g/100 g) in the diet increased VLDL apo B-100, indicating an increase in the number of VLDL particles. VLDL uptake by isolated hepatocytes and VLDL binding to hepatic membrane increased when the protein level increased from 10 to 20 g/100 g in the diet and decreased with 30 g/100 g protein, regardless of protein type. The dietary protein source did not affect plasma total cholesterol concentrations at any protein level. Feeding 20 g/100 g soybean protein compared with casein lowered plasma triglyceride concentrations and VLDL number as measured by decreased VLDL-protein, -phospholipid, -triglyceride, -cholesterol and -apo B-100. VLDL uptake by isolated hepatocytes and VLDL binding to hepatic membrane were higher in rats fed soybean protein than those fed casein. The higher VLDL uptake could be responsible for the hypotriglyceridemia in rats fed soybean protein.     

Baked cod consumption delayed the development of kidney and liver dysfunction and affected plasma amino acid concentrations,but did not affect blood pressure,blood glucose or liver triacylglycerol concentrations in obese fa/fa Zucker rats.

Obesity is associated with changes in amino acid metabolism, and studies show that ingestion of fish proteins influence amino acid composition in plasma and urine, in addition to affecting risk factors for metabolic syndrome. Since the majority of fish proteins consumed by humans are as fish fillet, it is of interest to investigate if cod fillet intake affects amino acid composition and metabolic disorders. We hypothesized that a modified AIN-93G diet containing cod fillet would affect amino acid compositions in plasma and urine in obese rats, and also affect risk factors for metabolic syndrome when compared to rats fed a regular AIN-93G diet with casein as the protein source. Obese Zucker fa/fa rats, a rat model of metabolic syndrome, received diets containing 25% protein from lyophilized baked cod fillet and 75% protein from casein (Baked cod diet), or a Control diet with casein for four weeks. The Baked cod diet affected the amino acid composition in plasma, with e.g., lower glycine, histidine, homoarginine, homocysteine, methionine, proline and tyrosine concentrations, but did not affect amino acid concentrations in urine. The concentrations of markers for kidney and liver dysfunction were lower in the Baked cod group, however blood pressure development, fasting and postprandial glucose, and hepatic triacylglycerol concentrations were similar to the Control group. To conclude, substituting 25% of dietary protein with baked cod fillet affected concentrations of some amino acids in plasma and delayed development of kidney and liver dysfunction, but did not affect blood pressure, glucose concentration or fatty liver.     

Digestibility of casein, formaldehyde-treated casein and soya-bean protein in relation to their effects on serum cholesterol in rabbits

Adult male rabbits were fed on semi-purified diets containing soya-bean protein isolate, casein or formaldehyde-treated casein as the protein source and 1 g cholesterol and 5 g of the non-absorbable marker chronic oxide/kg diet. The concentration of cholesterol in serum and in liver was increased on both the casein and formaldehyde-treated-casein diets. Excretion of bile acids and their concentration in faeces were lower in rabbits fed on casein or formaldehyde-treated casein when compared with rabbits fed on soya-bean protein. Apparent digestibility of nitrogen was lowest when formaldehyde-treated casein was fed, and highest on the casein diet. In rabbits fed on casein treated with formaldehyde, higher proportions of N were found in the water-soluble and trichloroacetic acid-insoluble protein fractions of the gastrointestinal tract contents compared with rabbits on the other two diets. Absorption of phosphate from the gastrointestinal tract was higher in rabbits fed on casein than in rabbits fed on soya-bean protein or formaldehyde-treated casein. The results indicate that, in rabbits, protein digestibility may not be an important determinant of serum cholesterol.