Biliary and fecal steroid excretion in rats fed partially hydrogenated soybean oil

Male Wistar rats were fed cholesterol-free or cholesterol-enriched diets containing partially hydrogenated soybean oil with different levels of trans-fatty acids or unhydrogenated soybean oil at the 10% level. The linoleic acid content of hydrogenated fat diets was adjusted to 3.6% of the total energy. Hydrogenated fat diets contained 29% and 41% trans-acids, mainly as t-18:1. Trans-fats exerted no untoward effects on growth parameters, but increased liver weight. Dietary hydrogenated fats influenced neither the concentration nor composition of biliary steroids, irrespective of the presence or absence of cholesterol in the diet. In rats fed a cholesterol-free diet, daily fecal output of neutral and acidic steroids was enhanced by hydrogenated fats and the magnitude of augmentation was proportional to the dietary level of trans-fatty acids. The increased fecal steroid excretion corresponded to an increase in total excreta. Hydrogenated fats also tended to enhance bile acid excretion when feeding a cholesterol-enriched diet. The results suggest that dietary trans-fatty acids, in relation to cis-polyunsaturated fatty acids, provoke demonstrable change in steroid homeodynamics.     

Contrasting effects of water-soluble and water-insoluble dietary fibers on bile acid conjugation and taurine metabolism in the rat

The effect of the type of dietary fiber on the bile acid and taurine metabolism was examined in rats. Diets containing 10% of various water-soluble fibers (citrus pectin, konjak mannan, guar gum) as compared to a fiber-free diet increased biliary excretion of total bile acids. In contrast, water-insoluble dietary fibers (cellulose, corn bran, chitin; 10% in the diets) as well as cholestyramine (5% in the diet) considerably, decreased bile acid excretion. Water-soluble dietary fibermediated increases in bile acid excretion were totally attributable to increases in glycine-conjugates. Thus, these fibers greatly increased by the bile acid glycine-to-taurine ratio (G/T). Excretio of glycine conjugates decreased more than that of taurine conjugates in rats fed various water-insoluble dietary fibers. As a results, G/T in rats fed water-insoluble fibers was significantly lowered as compared to G/T in animals fed a fiber-free diet. Cholestyramine did not affect the G/T ratio of bile acids. Fecal bile acid excretion and the activities of hepatic cholesterol 7α-hydroxylase (EC 1.14.13.17) in rats fed various water-soluble dietary fibers approximately doubled as compared to the respective values for rats fed a fiber-free diet. Whereas cholestyramine greatly increased these parameters, water-insoluble fibers did not significantly affect them. Various water-soluble fibers decreased hepatic concentration and urinary excretion of taurine as well as the activity of hepatic cysteine dioxygenase (EC 1.13.11.20). In contrast, water-insoluble fibers considerably increased hepatic taurine concentrations and enzyme activities. The parameters for taurine metabolism were unaffected by cholestyramine. It was suggested that the types of dietary fiber affected hepatic taurine synthesis and thus modified bile acid glycine/taurine ratios.     

Effect of chitosan feeding on intestinal bile acid metabolism in rats

The effect of chitosan feeding (for 21 days) on intestinal bile acids was studied in male rats. Serum cholesterol levels in rats fed a commercial diet low in cholesterol were decreased by chitosan supplementation. Chitosan inhibited the transformation of cholesterol to coprostanol without causing a qualitative change in fecal excretion of these neutral sterols. Increased fiber consumption did not increase fecal excretion of bile acids, but caused a marked change in fecal bile acid composition. Litcholic acid increased sigificantly, deoxycholic acid increased to a leasser extent, whereas hyodeoxycholic acid and the 6β-isomer and 5-epimeric 3α-hydroxy-6-keto-cholanoic acid(s) decreased. The pH in the cecum and colon became elevated by chitosan feeding which affected the conversion of primary bile acids to secondary bile acids in the large intestine. In the cecum, chitosan feeding increased the concentration of α-,β-, and ω-muricholic acids, and lithocholic acid. However, the levels of hyodeoxycholic acid and its 6β-isomer, of monohydroxy-monoketo-cholanoic acids, and of 3α, 6ξ, 7ξ-trihydroxy-cholanoic acid decreased. The data suggest that chitosan feeding affects the metabolism of intestinal bile acids in rats.     

Effects of rice bran oil on the intestinal microbiota and metabolism of isoflavones in adult mice

This study examined the effects of rice bran oil (RBO) on mouse intestinal microbiota and urinary isoflavonoids. Dietary RBO affects intestinal cholesterol absorption. Intestinal microbiota seem to play an important role in isoflavone metabolism. We hypothesized that dietary RBO changes the metabolism of isoflavonoids and intestinal microbiota in mice. Male mice were randomly divided into two groups: those fed a 0.05% daidzein with 10% RBO diet (RO group) and those fed a 0.05% daidzein with 10% lard control diet (LO group) for 30 days. Urinary amounts of daidzein and dihydrodaidzein were significantly lower in the RO group than in the LO group. The ratio of equol/daidzein was significantly higher in the RO group (p < 0.01) than in the LO group. The amount of fecal bile acids was significantly greater in the RO group than in the LO group. The composition of cecal microbiota differed between the RO and LO groups. The occupation ratios of Lactobacillales were significantly higher in the RO group (p < 0.05). Significant positive correlation (r = 0.591) was observed between the occupation ratios of Lactobacillales and fecal bile acid content of two dietary groups. This study suggests that dietary rice bran oil has the potential to affect the metabolism of daidzein by altering the metabolic activity of intestinal microbiota.     

Effect of excess dietary calcium on colon mucosal membranes and fecal lipids

The objective of the present studies was to examine the biochemical alterations in colon apical membranes upon feeding excess calcium to animals on a saturated fatty acid-rich diet. It has been suggested recently that excess dietary calcium may offer a protection to colon membranes as judged by histologic examination. Sprague-Dawley weanling male rats were fed a semisynthetic diet containing 14% beef fat plus 2% corn oil and either the calcium requirement or excess calcium in the form of calcium carbonate. Animals were fed the diets for 4 weeks. Feces were collected in the last 3 days. The results indicate that excess dietary calcium resulted in alteration in the density of 4 protein bands of colon apical membranes upon examination on SDS-gel electrophoresis. These bands contain 20% of membrane proteins. The diet had no effect on either the lipid content or fatty acid composition of the membranes. Excess dietary calcium resulted in 54% reduction in fecal water bile acids and a 44% reduction in fecal water free fatty acids. The reduction in fecal water lipids was due to alterations in the solubility of these lipids. This was not mediated through alterations in the pH of fecal water. The observed alterations in protein patterns of these membranes may be due to either the reduction of fecal water bile acids and free fatty acids or may be a direct effect of dietary calcium on membrane proteins.     

Effects of different levels of dietarytrans-octadecenoate on steroid metabolism in rats

Rats were fed semipurified diets containing olive oil or partially hydrogenated corn oil at the 5 or 20% level for ca. 30 days. These fat diets contained the same amount of octadecenoate but differed in the geometry with respect to each fat level. Contents oft-18∶1 were 26% and 41% of total fatty acids, respectively. The linoleic acid content was also made equivalent (3.8 energy %). After feeding on cholesterol-free diets, rats ontrans fat, compared to those oncis fat, showed: (a) no changes in serum cholesterol and apolipoprotein levels, (b) no effects on the bile flow and concentrations of biliary cholesterol or bile acids, (c) a trend toward increased fecal excretion of neutral and acidic steroids, (d) a lesser extent of transformation of cholesterol to coprostanol in the gut, and (e) no changes in the composition of biliary and fecal bile acids. Observations (c) and (d) were more marked with a hightrans fat regimen. These observations, except for serum apolipoproteins and fecal steroid excretion, were practically reproducible even when rats were fed cholesterol-enriched diets. A preliminary part of this study was presented at the 74th annual AOCS meeting, Chicago, 1983.     

Psyllium, not pectin or guar gum, alters lipoprotein and biliary bile acid composition and fecal sterol excretion in the hamster

Different soluble dietary fibers known to alter cholesterol metabolism were fed to golden Syrian hamsters, and their specific impact on lipoproteins, biliary bile acid profile, and fecal sterol excretion was evaluated. Semipurified diets containing 20% fat; 0.12% cholesterol; and 8% of psyllium (PSY); high (hePE) and low (lePE) esterified pectin; or high (hvGG) and low (lvGG) viscous guar gum were fed for 5 wk. Compared to control, PSY caused a significant reduction in plasma cholesterol (2.9±0.5 vs. 5.5±0.5 mmol/L), whereas hePE, lePE, hvGG, or lvGG had no apparent effect on plasma lipids. Hepatic total and esterified cholesterol were substantially decreased with PSY, pectin and guar gum, whereby PSY produced the most pronounced effect. Distinctive changes existed in the bile acid profile related to the different fibers. In contrast to pectin and guar gum, PSY caused a significant increase in the cholate:chenodeoxycholate and the glycine:taurine conjugation ratio. Pectin and guar gum did not alter daily fecal neutral sterol excretion while PSY caused a 90% increase due to a higher fecal output. Daily fecal bile acid excretion and total fecal bile acid concentration were significantly increased by PSY, whereas hePE, lePE, hvGG, and lvGG revealed no or only minor effects. Taken together, the disparate hypocholesterolemic effects of PSY, pectin, and guar gum on cholesterol and bile acid metabolism in the hamster are possibly related to different physicochemical properties, e.g., viscosity and susceptibility to fermentation, affecting the fiber-mediated action in the intestine.     

Highly hydrogenated dietary soybean oil modifies the responses to polychlorinated biphenyls in rats

The effects of dietary highly hydrogenated soybean oil (HSO) upon the changes caused by dietary polychlorinated biphenyls (PCBs) were examined in rats. Six groups of rats were fed the following diets for 30 d: a 20% soybean oil-containing diet (control diet), a diet in which a half of soybean oil was substituted with HSO (HSO-A diet), a diet in which cellulose powder was replaced with HSO (HSO-B diet) and these diets supplemented with 100 ppm PCBs (control+PCBs, HSO-A+PCBs and HSO-B+PCBs diets). Hepatic concentration of PCBs and relative liver weight were markedly decreased in rats fed with the HSO-A+PCBs diet compared with those fed with the other diets containing PCBs. Liver lipids and liver cholesterol were considerably decreased with a reciprocal increase in fecal sterol excretion by rats fed the HSO-A+PCBs and the HSO-B+PCBs diets compared with those fed with the control+PCBs diet. The fatty acid composition in hepatic phospholipids showed an independent increase of the saturated fatty acid content induced by dietary HSO and PCBs. Dietary PCBs also caused decreases in the amounts of monounsaturated and n-3 polyunsaturated fatty acids. These results suggest that dietary HSO prevents accumulation of PCBs in the liver and promotes the excretion of lipids stimulated by PCBs, accompanied by a change in fatty acid metabolism.     

Effect of cholestyramine on bile acid metabolism in conventional rats

Effects of cholestyramine on biliary secretion of cholesterol, phospholipids and bile acids and fecal excretion of sterols and bile acids were examined in Wistar male rats. Six rats were fed a basal diet, and the other six were fed a basal diet supplemented with 5% cholestyramine for eight days. Bile flow and biliary secretion of bile acids and phospholipids (per hour per rat) decreased with cholestyramine treatment, while biliary cholesterol secretion (per hour per rat) remained unchanged. In the biliary bile acid composition, a marked increase of chenodeoxycholic acid with a concomitant decrease of β-muricholic acid was observed in cholestyramine-treated rats. Fecal excretion of total sterols and bile acids increased about three-and four-fold, respectively, after cholestyramine treatment. The increase of fecal bile acids derived from cholic acid was more predominant than that derived from chenodeoxylcholic acid, resulting in an increase of the cholic acid group/chenodeoxycholic acid group ratio.     

Cholestyramine alters the lipid and energy metabolism of chicks fed dietary medium-or long-chain triacylglycerol

The effects of cholestyramine, a bile acid binding polymer, on the lipid and energy metabolism of chicks given dietary medium-chain triacylglycerol (MCT) or long-chain triacylglycerol (LCT) were investigated. Chicks (from 8 to 17 days of age) were fed diets containing MCT or LCT at 200 g oil/kg diet with or without 2% cholestyramine under equalized feeding conditions. An adjusted LCT diet was formulated in order to supply another group with daily nutrients and dietary metabolizable energy (ME) equal to MCT groups, except for corn starch. ME intakes of chicks given MCT or LCT diets were reduced by cholestyramine; consequently, fat and energy retention was reduced, though the reduction was more drastic in chicks fed LCT. This was caused by a change in amounts of the fecal excretion of fat and bile acids. Cholestyramine enhanced the excretion of octanoic acid (8∶0) in the feces, which suggests that bile acids are needed for 8∶0 absorption. Cholestyramine affects the utilization of dietary MCT and LCT by lowering fat and energy retention in chicks. However, the effect of cholestyramine on MCT utilization was smaller than its effect on utilization of LCT.     

Effects of dietaryTrans-fat on biliary and fecal steroid excretion and serum lipoproteins in rats

Rats were fed cholesterol-free or cholesterol-enriched diets containing olive oil or partially hydrogenated corn oil at the 10% level for ca. 30 days (c-18∶1, 77.0% in the former diet andc-18∶1, 24.7% andt-18∶1, 42.5% in the latter). The linoleic acid content of these fat diets was made equivalent (1.7 energy%). After feeding cholesterol-free diets,trans fat compared tocis fat showed(a) no untoward effects on growth parameters, (b) a reduction of serum cholesterol levels without influencing concentrations of serum apolipoproteins A-I, B and E, (c) no effects on the bile flow and the concentration of biliary cholesterol and bile acids, (d) an increasing trend of fecal excretion of neutral and acidic steroids, both in terms of mg/day and mg/g feces, and (e) rather equivocal change in the composition of fecal, but not biliary steroids. Similar response patterns were also observed when cholesterol-enriched diets were fed except for a decrease in serum apo B and an ineffectiveness to increase fecal acidic steroids. Together with the results obtained from experiments simultaneously performed with safflower oil and completely hydrogenated corn oil, it seems that the steroid metabolism can be specificallymodified by the geometry of dietary fats.     

Distribution of bile acids in rats

Distribution and biliary and fecal excretion of bile acids were examined in Wistar strain male rats of about 300 g body weight. The pool size of the rats on ordinary diet was 40 mg/rat, biliary secretion was 14 mg/hr, and fecal excretion was 10 mg/day. Bile acids were mainly located in the small and large intestinal contents, 87% and 10%, respectively; but a portion was found in the intestinal wall and the liver. Rats fed 2% cholesterol-supplemented diet for a week showed similar values for pool size and biliary secretion with the rats on ordinary diet, but higher values for fecal excretion and distribution ratio in the large intestinal contents. Cholic acid was a major component in the bile, small intestinal wall, small intestinal content and liver, while the bile acid composition ratios were roughly similar to each other, although a relatively large amount of α-muricholic acid was found in the intentinal wall and liver. Both the wall and content compositions of the large intestine were similar to that of the feces, in which lithocholic, deoxycholic, α- and β-muricholic acids were the main components, although the ratios of α- and β-muricholic acids in the large intestinal wall were larger than those in the intestinal contents or feces. The high concentrations of these bile acids may indicate a difference of transport velocity across the cell membrane, but the mechanism is not known.     

Effect of dietary polyunsaturated pork on plasma lipids and sterol excretion in man

Pork, enriched in linoleic acid content, was compared with conventional pork in the diet of three human subjects with respect to the plasma cholesterol concentration and the excretion in feces of neutral sterols and bile acids. Since the fatty acids in pork glyceride have an unusual positional distribution, the redistribution that might occur during the absorption and disposition of a fat meal was also studied. The plasma cholesterol was lower with polyunsaturated pork, the difference, 14 mg/100 ml plasma, being of the order expected from the change in polyunsaturated to saturated fatty acid ratio. On average, the excretion of neutral sterols was 57% greater with polyunsaturated than with conventional pork in all three subjects, and in this respect the results resembled the findings with polyunsaturated ruminant fats. During the absorption of pork fat, the high proportion of palmitate in the 2 position of lard triglyceride served as a useful marker, since human triglyceride carries mainly unsaturated fatty acids in that position. There were stepwise changes in the fatty acid composition at the 2 position of triglyceride as the fat was absorbed, transported through, and cleared from plasma, the palmitate being gradually replaced by oleate and linoleate. By contrast, the total fatty acid profile in the triglyceride changed relatively little, implying selective reacylation with palmitate at the 1 and/or 3 position. During the clearing of dietary triglyceride, the porcine triglyceride was thus converted to the form occurring in humans.     

Atorvastatin Modulates Bile Acid Homeostasis in Mice with Diet-Induced Nonalcoholic Steatohepatitis

Bile acids (BA) play a significant role in the pathophysiology of nonalcoholic steatohepatitis (NASH). The present study evaluates the modulation of bile acid metabolomics by atorvastatin, a cholesterol-lowering agent commonly used to treat cardiovascular complications accompanying NASH. NASH was induced in mice by 24 weeks of consuming a high–saturated fat, high-fructose, and high-cholesterol diet (F), with atorvastatin administered orally (20 mg/kg/day) during the last three weeks. Biochemical and histological analyses confirmed the effectiveness of the F diet in inducing NASH. Untreated NASH animals had significantly reduced biliary secretion of BA and increased fecal excretion of BA via decreased apical sodium-dependent bile salt transporter (Asbt)-mediated reabsorption. Atorvastatin decreased liver steatosis and inflammation in NASH animals consistently with a reduction in crucial lipogenic enzyme stearoyl–coenzyme A (CoA) desaturase-1 and nuclear factor kappa light chain enhancer of activated B-cell pro-inflammatory signaling, respectively. In this group, atorvastatin also uniformly enhanced plasma concentration, biliary secretion and fecal excretion of the secondary BA, deoxycholic acid (DCA). However, in the chow diet–fed animals, atorvastatin decreased plasma concentrations of BA, and reduced BA biliary secretions. These changes stemmed primarily from the increased fecal excretion of BA resulting from the reduced Asbt-mediated BA reabsorption in the ileum and suppression of synthesis in the liver. In conclusion, our results reveal that atorvastatin significantly modulates BA metabolomics by altering their intestinal processing and liver synthesis in control and NASH mice.     

Bile acid metabolism in young-old parabiotic rats

Serum cholesterol, triglyceride and phospholipid levels, liver cholesterol concentration, bile flow, biliary cholesterol, phospholipid and bile acid secretion rates, fecal sterol and bile acid levels and their bile acid compositions were examined in young-old parabiotic rats and compared with those in young and old control rats and young-young parabiotic rats. Bile acid composition was expressed in terms of the cholic acid group/chenodeoxycholic acid group (CA/CDCA) ratio. Body weight (BW) gain decreased after parabiosis especially in old rats, but the liver weight (g/100 g BW), diet-intake, feces dry weight, liver cholesterol concentration and fecal sterol level were almost the same in all the groups. The biliary bile acid secretion rate was higher and the fecal bile acid level was lower in old rats than those in young rats but both the levels became comparable with those in young rats after parabiosis of old rats with young rats. Young rats, however, showed no changes in these levels after parabiosis. The serum cholesterol level and the biliary and fecal CA/CDCA ratios in old rats were higher than those in young rats but decreased after parabiosis with young rats, although they were still higher than those in young rats. The serum cholesterol level in young rats increased after parabiosis with old rats, but not after parabiosis with young rats, and the fecal bile acid level and the CA/CDCA ratio were not changed in either case. It is concluded from these findings that the serum cholesterol level and the CA/CDCA ratio increased with age and that these increases were prevented after parabiosis with young rats, while young rats, although their serum cholesterol level was increased, showed no increase in the CA/CDCA ratio after parabiosis with old rats.     

Analysis of fecal neutral steroids and bile acids in humans on constant fat diet

A method is described for the separation and quantification of fecal neutral steroids and fecal bile acids. The fecal extract is separated into the neutral steroid fraction and bile acid fraction with ionexchange resin columns. The principal neutral steroids and bile acids are then separated and quantitated by thin-layer chromatography. Values for the fecal neutral steroids, cholesterol, coprostanol and coprostanone and fecal bile acids, deoxycholic acid and lithocholic acid of 5 subjects on a constant fat diet for a 3-week period are presented.     

Dietary fat composition influences fatty acid composition of milk fat globule membrane in lactating cows

Milk fat globule membranes are derived directly from the apical plasma membrane of mammary epithelial cells. To evaluate the effect of dietary fat on mammary membranes, we determined the fatty acid composition of the milk fat globule membrane in lactating dairy cows fed diets supplemented with fats of different fatty acid composition, or infused intravenously with soy oil emulsion. A preliminary survey, using an abbreviated preparation procedure (membranes isolated at 48,000 x g-max for 15 min), yielded about 45% of the total membrane fatty acids that could be recovered by centrifuging at the same speed for 120 min, and showed that changes in fatty acid composition of membranes reflected dietary fatty acids to some extent. Dietary palmitic acid increased the content of 16:0 in the membranes. A high corn diet increased ruminal formation of t18:1, and its level increased to 12% of membrane fatty acids. Infusion of soy oil emulsion increased 18:2 membrane content, and decreased the levels of 18:1 and 20:4. All treatments decreased the ratio of unsaturated/saturated fatty acids as compared to controls, whereas the ratio of polyunsaturated/saturated fatty acids was increased by feeding a high corn diet or by infusing soy oil. The ratio of 18:2/c18:1 increased from 0.31 to 1.0 after infusing soy oil for 4 days. The fatty acids of membranes isolated upon 120-min centrifugation were slightly more saturated. The differences were not sufficiently large, however, to affect overall results significantly.     

Influence of dietary fiber on bile acid metabolism

Fiber, when fed to animals or man, will generally cause increased excretion of bile acids. The level of bile acid excretion appears to be a function of the structure of the fiber. Fiber binds bile acids and bile salts in vitro. The extent of binding is characteristic for each type of fiber and each substrate. Bile acid binding may be one mechanism of the physiological action of fiber.     

Effects of a single ingestion of sodium taurocholate on esterified cholesterol concentration in liver and cholesterol turnover in the rat

The overall response of the rat’s cholesterol metabolism to a single ingestion of taurocholate (80 mg) was studied with the isotopic equilibrium method. The bile acid production, measured by the daily¹⁴CO₂ output of rats in isotopic equilibrium of [26-¹⁴C]-cholesterol, initially decreased and then increased. Conversely, the hepatic concentration of esterified cholesterol first increased and then decreased. Moreover, the ingestion of taurocholate increasing the intestinal absorption coefficient of dietary cholesterol increased the abosprtion and decreased the fecal excretion and the intestinal biosynthesis of cholesterol. The balance of these last effects is an excess cholesterol inflow. The classical hypothesis of negative feedback regulation of bile acid production fails to explain the observed biphasic effect of taurocholate. This compound, when its origin is exogenous, appears to stimulate the storage of esterified cholesterol in the liver, at the expense of bile acid synthesis. This accumulation rate takes into account not only the decrease in cholesterol transformation into bile acids but also the excess inflow of cholesterol. As the exogenous taurocholate was eliminated from the body, cholesteryl ester hydrolysis occurred and provided a supplementary source of free cholesterol for bile acid synthesis.