Daily Fecal Bile Acid Excretion Research Articles

Screening for Cholesterol-Lowering Probiotics from Lactic Acid Bacteria Isolated from Corn Silage Based on Three Hypothesized Pathways.

A total of 85 strains of lactic acid bacteria were isolated from corn silage in this study and analyzed in vitro for their cholesterol removal, NPC1L1 protein down-regulation and bile salt deconjugation ability, respectively. Nineteen strains were selected for further analysis for their probiotic potential. Finally, 3 strains showing better probiotic potential were evaluated for their cholesterol-lowering activity in hamsters. The strains showing the greater cholesterol removal and NPC1L1 protein down-regulation activity had no significant effects on serum and hepatic cholesterol levels in hamsters (p > 0.05). However, Lactobacillus plantarum CAAS 18008 (1 × 109 CFU/d) showing the greater bile salt deconjugation ability significantly reduced serum low-density lipoprotein cholesterol, total cholesterol, and hepatic total cholesterol levels by 28.8%, 21.7%, and 30.9%, respectively (p < 0.05). The cholesterol-lowering mechanism was attributed to its bile salt hydrolase activity, which enhanced daily fecal bile acid excretion levels and thereby accelerated new bile acid synthesis from cholesterol in liver. This study demonstrated that the strains showing greater cholesterol removal and NPC1L1 protein down-regulation activity in vitro hardly reveal cholesterol-lowering activity in vivo, whereas the strains showing greater bile salt deconjugation ability in vitro has large potential to decrease serum cholesterol levels in vivo.

Effect of esterified 4-desmethylsterols and -stanols or 4,4'-dimethylsterols on cholesterol and bile acid metabolism in hamsters.

4-Desmethylsterols and -stanols reduce plasma total cholesterol (TC) and LDL cholesterol by inhibition of intestinal cholesterol absorption, while the cholesterol-lowering potential of 4,4'-dimethylsterols is less well defined. The present study aimed to compare the effects of 4-desmethylsterols, -stanols, and 4,4'-dimethylsterols on plasma and hepatic cholesterol, sterol excretion and bile acid metabolism. Male golden Syrian hamsters were fed diets containing 13 g/100 g fat, 008 g/100 g cholesterol and 0 (control), 0.24 or 0.48% (w/w) esterified 4-desmethylsterols (sterols) and esterified hydrogenated 4-desmethylsterols (stanols) from common vegetable oils or esterified 4,4'-dimethylsterols from rice bran oil for 5 weeks. Sterol and stanol esters at the dose of 0.24% were equally effective and significantly (P<0.05) lowered TC by 15%, while 0.24% 4,4-dimethylsterols reduced TC by 10%. Liver total and esterified cholesterol concentrations were significantly (P<0.05) lowered by 40, 22, 43 and 31% in hamsters fed 0.48% sterols, 0.24% stanols, 0.48% stanols or 0.48% dimethylsterols, respectively. Daily faecal bile acid excretion and hepatic cholesterol 7alpha-hydroxylase activity were not altered, indicating that sterols, stanols and dimethylsterols had no effect on the intestinal re-absorption of bile acids or on hepatic bile acid synthesis. Daily excretion of cholesterol was significantly higher in hamsters fed esterified sterols and stanols, but was only slightly increased in those fed dimethylsterols. The results indicate that esterified sterols and stanols were equally effective in lowering plasma TC and LDL cholesterol, while dimethylsterol esters caused a weaker cholesterol-lowering effect. Sterols and stanols achieve their cholesterol-lowering effect by stimulating faecal cholesterol excretion through inhibiting intestinal cholesterol absorption, but do not affect bile acid excretion. Other mechanisms need to be considered to explain the effect on plasma and hepatic cholesterol of dimethylsterols.

Effect of esterified 4-desmethylsterols and -stanols or 4,4′-dimethylsterols on cholesterol and bile acid metabolism in hamsters

4-Desmethylsterols and -stanols reduce plasma total cholesterol (TC) and LDL cholesterol by inhibition of intestinal cholesterol absorption, while the cholesterol-lowering potential of 4,4′-dimethylsterols is less well defined. The present study aimed to compare the effects of 4-desmethylsterols, -stanols, and 4,4′-dimethylsterols on plasma and hepatic cholesterol, sterol excretion and bile acid metabolism. Male golden Syrian hamsters were fed diets containing 13 g/100 g fat, 0·08 g/100 g cholesterol and 0 (control), 0·24 or 0·48 % (w/w) esterified 4-desmethylsterols (sterols) and esterified hydrogenated 4-desmethylsterols (stanols) from common vegetable oils or esterified 4,4′-dimethylsterols from rice bran oil for 5 weeks. Sterol and stanol esters at the dose of 0·24 % were equally effective and significantly (P&lt;0·05) lowered TC by 15 %, while 0·24 % 4,4-dimethylsterols reduced TC by 10 %. Liver total and esterified cholesterol concentrations were significantly (P&lt;0·05) lowered by 40, 22, 43 and 31 % in hamsters fed 0·48 % sterols, 0·24 % stanols, 0·48 % stanols or 0·48 % dimethylsterols, respectively. Daily faecal bile acid excretion and hepatic cholesterol 7α-hydroxylase activity were not altered, indicating that sterols, stanols and dimethylsterols had no effect on the intestinal re-absorption of bile acids or on hepatic bile acid synthesis. Daily excretion of cholesterol was significantly higher in hamsters fed esterified sterols and stanols, but was only slightly increased in those fed dimethylsterols. The results indicate that esterified sterols and stanols were equally effective in lowering plasma TC and LDL cholesterol, while dimethylsterol esters caused a weaker cholesterol-lowering effect. Sterols and stanols achieve their cholesterol-lowering effect by stimulating faecal cholesterol excretion through inhibiting intestinal cholesterol absorption, but do not affect bile acid excretion. Other mechanisms need to be considered to explain the effect on plasma and hepatic cholesterol of dimethylsterols.

Cholesterol Reduction by Glucomannan and Chitosan Is Mediated by Changes in Cholesterol Absorption and Bile Acid and Fat Excretion in Rats

Glucomannan, a viscous polysaccharide, and chitosan, a derivative of chitin, have both been demonstrated to lower cholesterol in animals. However, the mechanism of cholesterol lowering has not been established for either material. This study was conducted to determine the effect of glucomannan (G), chitosan (CH), or an equal mixture of the two (G + CH) on cholesterol absorption and fat and bile acid excretion. Rats were fed a modified AIN-93G diet for 18 d containing 0.125 g/100 g cholesterol and initially 10 g/100 g of the test materials or cellulose (C) as the control. However, the concentration of test materials and cellulose was reduced to 7.5 g/100 g after 1 wk due to lower weight gain compared with controls. Total liver cholesterol was significantly reduced in G, CH and G + CH groups compared with the C group. The intestinal contents supernatant viscosity of the C and the CH groups was negligible, whereas both G and G + CH produced high viscosities. Cholesterol absorption, measured by the fecal isotope ratio method, was significantly reduced from 37.5% in the C group to 20.2% in G, 18.2% in G + CH and 9.4% in CH. Daily fecal fat excretion did not differ between the C and G groups, but was significantly greater in G + CH and CH compared with the C and G groups. Daily fecal bile acid excretion was significantly greater in the CH and G + CH groups compared with the C and G groups. These results suggest that G lowered liver cholesterol by a viscosity-mediated interference of cholesterol absorption. In contrast, CH appears to lower cholesterol through a different mechanism.

Increased Fecal Bile Acid Excretion and Changes in the Circulating Bile Acid Pool Are Involved in the Hypocholesterolemic and Gallstone-Preventive Actions of Psyllium in Hamsters

The lipid-lowering effect of psyllium (PSY) is well established. Enhanced fecal bile acid excretion and a stimulation of hepatic bile acid synthesis are discussed as primary mechanisms of this action. To further examine the effect of bile acid excretion and specifically of compositional alterations in the bile acid pool on the cholesterol-lowering and gallstone-preventing action of PSY, male golden Syrian hamsters were fed lithogenic diets containing 5 g/100 g fat, 0.4 g/100 g cholesterol and 0 (control), 4 or 6% PSY or 1% cholestyramine (CHY). PSY significantly lowered plasma total cholesterol and triacylglycerol at a magnitude comparable to that induced by CHY. Although hepatic cholesteryl ester accumulation was completely inhibited by CHY, PSY did not prevent the hepatic storage of esterified cholesterol. PSY and CHY caused distinct alterations in the bile acid profile. PSY caused a selective reduction of taurine-conjugated bile acids, especially of taurochenodeoxycholate. As a result, the glycine:taurine conjugation and the cholate:chenodeoxycholate ratios were significantly higher in PSY-fed hamsters. PSY and CHY normalized the lithogenic index and prevented cholesterol gallstone formation compared with controls. Daily fecal bile acid excretion was ∼400% greater in hamsters fed 6% PSY, whereas CHY caused an 11-fold increase. Daily neutral sterol excretion did not differ in PSY-fed hamsters but was >100% greater in those fed CHY than in controls. These data emphasize the potent lipid-lowering effect of PSY. Increased fecal bile acid excretion and alterations of the circulating bile acid pool by removal of dihydroxy bile acids (e.g., taurochenodeoxycholate) appear to be main modulators of the hypocholesterolemic action of PSY by leading to an up-regulation of hepatic bile acid synthesis.

Dietary Inulin Lowers Plasma Cholesterol and Triacylglycerol and Alters Biliary Bile Acid Profile in Hamsters

The mechanisms by which inulin may elicit its lipid-lowering effect are not well elucidated. To examine the lipid-lowering potential of inulin and especially its effect on bile acid metabolism, male golden Syrian hamsters were fed semipurified diets containing 20 g/100 g fat, 0.12 g/100 g cholesterol and 0 (control), 8, 12 or 16% inulin for 5 wk. Plasma total cholesterol concentrations were significantly lowered by 18, 15 and 29% in hamsters fed 8, 12 and 16% inulin, respectively. Dietary inulin specifically decreased VLDL cholesterol, which was significantly lower in hamsters fed 16% inulin compared with controls (1.1 ± 0.3 vs. 2.9 ± 0.6 mmol/L). LDL and HDL cholesterol were not significantly affected by dietary inulin. Plasma triacylglycerol was significantly reduced by 40 and 63% in hamsters fed 12 and 16% inulin, respectively. Hepatic total cholesterol and particularly esterified cholesterol accumulation were significantly lower in hamsters fed 8% inulin compared with controls. All three levels of dietary inulin caused distinct alterations in the bile acid profile of gallbladder bile. Taurochenodeoxycholic acid was significantly lower, whereas glycocholic and glycodeoxycholic acid were greater in hamsters fed inulin. Daily fecal bile acid excretion (μmol/d) tended to be greater (P = 0.056) in inulin-fed hamsters compared with controls, whereas daily neutral sterol excretion was not affected. These data demonstrate that the lipid-lowering action of inulin is possibly due to several mechanisms, including altered hepatic triacylglycerol synthesis and VLDL secretion and impaired reabsorption of circulating bile acids.

Dietary Stearic Acid Reduces Plasma and Hepatic Cholesterol Concentrations without Increasing Bile Acid Excretion in Cholesterol-Fed Hamsters1–4

Although there is general agreement that saturated fatty acids elevate plasma cholesterol concentrations, the relative effects of individual fatty acids on cholesterol and bile acid metabolism are less clear. In this study, cholesterol and bile acid responses to diets enriched in different saturated fatty acids were investigated in hamsters. The six diets examined were as follows: 5% fat (g/100 g) enriched in palmitic acid (16:0) with no cholesterol, 5% fat 16:0-enriched, 0.05% cholesterol (wt/wt), and four diets containing 0.05% cholesterol and 15% fat with each diet enriched in lauric (12:0), myristic (14:0), palmitic (16:0), or stearic acid (18:0). Total plasma cholesterol concentration was significantly greater in hamsters fed the 14:0-enriched diet relative to those fed the 18:0-enriched diet (P < 0.05). Both plasma and liver cholesterol concentrations of hamsters fed 18:0 did not differ from those of the group fed no dietary cholesterol. In all instances, differences in total plasma cholesterol were accounted for within the HDL fraction; no significant treatment differences in VLDL or LDL cholesterol were found. Total daily fecal bile acid excretion was higher in hamsters fed the 15% fat 16:0 diet compared with those fed no dietary cholesterol (P < 0.05), but not significantly different from other treatment groups. There was greater deoxycholic acid excretion (P < 0.05) from hamsters fed the 14:0 and 16:0 diets compared with those fed the 18:0-enriched diet. Small intestinal + gallbladder bile acids, an index of pool size, did not differ significantly among the groups. The observed relative hypocholesterolemic effect of stearic acid was not mediated by increased bile acid excretion.

Metabolic and morphometric changes in small and large intestine in rats fed high-fiber diets.

Structural modifications of the intestine following fiber intake may play an important role in the physiological effects induced by dietary fiber. The aim of this study was to examine possible mechanisms for the hypocholesterolemic effects of dietary fibers by evaluating both biochemical and morphometric parameters. Thirty male Sprague-Dawley rats were divided into 3 groups and fed either a fiber-free, 15% fiber high methoxy pectin, or 15% cellulose diet for a 4-wk period. Plasma cholesterol levels, hepatic enzyme activity, and fecal bile acid excretion were determined, and changes in the intestinal structure were evaluated. Daily fecal bile acid excretion was significantly higher in rats fed the pectin diet accompanied by elevated HMG-CoA reductase activity. Pectin feeding led to significant elongation of both the small and large intestine whereas cellulose only affected colon length. Computerized image analysis of intestinal cross-sections showed enlarged muscle area in the ileum and midcolon of pectin-fed rats and greater mucosal area in the colon. Circular muscle cell hypertrophy was also observed in the colon length. Computerized image analysis of intestinal cross-sections showed enlarged muscle area in the ileum and midcolon of pectin-fed rats and greater mucosal area in the colon. Circular muscle cell hypertrophy was also observed in the colon. In contrast, the effects of cellulose feeding were limited to elongation of the colon accompanied by muscle cell hypertrophy, and little effect on ileal morphology was documented. These data indicate that the hypocholestrolemic effects of pectin may be due to increased bile acid secretion accompanied by significant morphological changes in the ileum and colon.

Faecal bile acid excretion during detoxification in patients with alcohol abuse.

We wanted to ascertain whether alcohol abuse is associated with a changed faecal bile acid excretion. Faecal bile acid excretion was studied in seven chronic alcoholic subjects when admitted to hospital for detoxification. Bile acids in faeces and serum were quantified by gas-liquid chromatography and identified by mass spectrometry. Daily faecal bile acid excretion was on average 948 mumol in the patients at admission, compared with 400 mumol in eight healthy controls (P < 0.01). The daily faecal bile acid excretion decreased slowly during the detoxification period but was still higher in the patients than in controls after 9 days of detoxification (P < 0.05). Faecal bile acid excretion varied greatly between the different patients, with poor correlation to diarrhoea. In one patient the faecal bile acid excretion was remarkably high, amounting to an average of 6800 mumol day-1 during the first 3 days. The percentage metabolites of total faecal bile acids in the patients did not differ significantly from that of controls. The faecal wet weight was higher in the patients than in the controls, although significantly so only after the first 3 days of detoxification. Faecal bile acid excretion is increased after alcohol abuse. A gradual decrease in faecal bile acid excretion was observed during the 9-day observation period when the subjects abstained from alcohol.

Serum lipid and fecal bile acid changes with cereal, vegetable, and sugar-beet fiber feeding

Thirty-four subjects consumed six controlled formula diets for 3 wk each, supplemented with 0 g added fiber, 10 and 30 g dietary fiber as wheat bran (WB), 10 and 30 g dietary fiber as mixed vegetable fiber (VF), and 30 g dietary fiber as sugar-beet fiber (SBF). Serum cholesterol changes for fiber free, 10 g WB, 30 g WB, 10 g VF, 30 g VF, and 30 g SBF (-0.13, -0.18, -0.05, -0.17, -0.24, and -0.70 mmol/L, respectively) were significant for 30 g VF and 30 g SBF. Reduction in total cholesterol with SBF was largely due to significant lowering of low-density-lipoprotein cholesterol. Total fecal bile acid concentrations were significantly higher with the fiber-free diet than with 30 g WB, VF, and SBF (P less than 0.001) and were also higher with 30 g SBF than with 30 g WB and 30 g VF (P less than 0.005). Daily fecal bile acid excretion was not different on 30 g SBF compared with 30 g WB and 30 g VF. Differences in cholesterol reduction across the diets could not be explained by differences in fecal bile acid excretion.

Experimental colonic carcinogenesis: changes in faecal bile acids after promotion of intestinal tumours by small bowel resection in the rat.

Small bowel resection promotes the development of colonic tumours in azoxymethane treated rats. As high faecal bile acid concentrations are associated with colonic cancer and may be altered by resection, we have studied changes in faecal bile acid concentrations during promotion of colonic carcinogenesis by increasing small bowel resection. Twenty rats in each group underwent either jejunal transection or 20%, 50%, or 80% proximal small bowel resection. Tumours were induced with azoxymethane 10 mg/kg by 12 weekly subcutaneous injections, and faecal bile acid concentrations were measured at six and 16 weeks. Colonic tumour number rose from 0.6 per rat in the transection group to 1.6 per rat in the 50% resection group (p less than 0.01) but were not significantly different to transection values at 0.8 per rat in the 80% resection group. Total daily faecal bile acid excretion and bile acid concentrations fell with increasing resection from 14.2 (1.6) mg/rat/day and 5.8 (0.7) mg/g dry faeces respectively in the transection group to 6.5 (0.5) mg/rat/day and 2.9 (0.2) mg/g respectively in the 80% resection group (p less than 0.001). The greatest reduction was seen in the concentration of deoxycholic acid which has been particularly associated with the aetiology of colonic cancer. The promotion of colonic tumours following small bowel resection in carcinogen treated rats is unlikely to be mediated by changes in faecal bile acid concentration or composition.

Effect of bile acid conjugation pattern on bile acid metabolism in normal humans

Six male subjects were fed taurine 0.5 g six times daily for 2 wk to determine the effect of a shift in bile acid conjugation pattern upon bile acid metabolism. Duodenal bile acids were analyzed, and bile acid pool size, daily fecal excretion, and biliary excretion rate were quantified. In addition, daily biliary excretion rate of cholesterol and phospholipid were quantified, and biliary saturation with cholesterol was estimated. The dose of taurine caused reversal of the bile acid glycine-to-taurine conjugation ratio. Total bile acid pool size decreased, as did the pool size of chenodeoxycholic acid. Pool sizes of cholic and deoxycholic acids did not change. Daily fecal bile acid excretion decreased slightly. Biliary secretion rates of cholesterol, phospholipid, and bile acids did not change, nor did biliary cholesterol saturation. Pool size can decrease because of increased bile acid catabolism or decreased synthesis. The fact that bile acid excretion failed to increase, and actually decreased slightly, suggests that the effect is upon bile acid synthesis. In normal humans, the effect is small and probably physiologically unimportant. In special cases, however, such as during ursodeoxycholic acid therapy, the effect of shifting conjugation pattern may become important.

A modified assay of total faecal bile acid excretion for clinical studies

Faecal bile acid excretion was estimated in a clinical laboratory using gas liquid chromatography. The original method of Grundy et al was simplified so that quantitative thin layer chromatography was not required. Rigid dietary control or inpatient facilities were unnecessary for clinical studies. Control patients studied under these conditions had a daily faecal bile acid excretion of 0.37 +/- 0.13 g/day. Patients with ileal resections varied markedly with bile acid excretions ranging from 0.25--6.32 g/day (mean 2.36 +/- 1.11).

Fecal excretion of bile acids: a new technique for studying bile acid kinetics in patients with ileal resection.

The fecal elimination and enterohepatic circulation of bile acid was studied in 11 patients. 10 patients with varying degrees of ileal disease or resection and 1 patient with pancreatic insufficiency and no ileal disease. A new technique was employed which involved the nearly simultaneous administration of cholic acid-(14)C and a nonabsorbable marker. (51)CrCl(3). Each individual stool specimen was collected for 36-96 hr and analyzed separately. Assay of the radioactivity of each isotope allowed the accurate determination of an excretion rate for both cholic acid and (51)Cr. The difference between these rates was used to calculate an absorption coefficient for cholic acid. In addition, bile acid concentration measured by the steroid dehydrogenase technique, and the water content of each stool was determined. THE PATIENTS WERE DIVIDED INTO GROUPS DEPENDING UPON HOW MUCH SMALL INTESTINE WAS RESECTED OR DISEASED: six patients with less than 100 cm of ileal resection or disease (group A), and five patients with more than 100 cm of ileal disease or resection (group B). The (51)Cr excretion rate was similar in the two groups, but cholic acid-(24)C excretion rates were significantly more rapid in group B than in group A. The cholic acid absorption coefficient was essentially normal in the patient with pancreatic insufficiency, moderately decreased in group A patients, and extremely low or zero in group B patients. It was inversely related to the length of intestine diseased or resected. Daily fecal bile acid excretion was normal to twice normal in group A patients and 2-8 times normal in group B patients. In all patients with ileal disease or resection, there was a direct correlation between fecal bile acid, fecal mass, and fecal water. Each millimole of additional bile acid in the stool was associated with an increase in stool water of 11 moles (P < 0.01). These studies show that the kinetics of bile acids in the enterohepatic circulation can be accurately studied in patients with extensive ileal resection. The regular relationship between fecal bile acid and fecal mass and water suggests, but does not prove, a critical role of bile acid in determining stool water.