phytosterols and stigmastanol

Foods with added phytosterols/phytostanols (PS) are recommended to lower LDL cholesterol (LDL-c) concentrations. Manufacturers have incorporated PS into a variety of common foods. Understanding the cholesterol-lowering impact of the food matrix and the PS characteristics would maximize their success and increase the benefit to consumers. This review systematically examines whether the PS characteristics and the fatty acid composition of foods with added PS affects serum LDL-c. A total of 33 studies published between the years 1998 and 2011 inclusive of 66 individual primary variables (strata) were evaluated. The functional food matrices included margarine, mayonnaise, yogurt, milk, cheese, meat, grain, juice, and chocolate. Consistently, ≥10% reductions in LDL-c were reported when the characteristics of the food matrix included poly- and monounsaturated fatty acids known to lower LDL-c. Also, >10% mean reductions in LDL-c were reported when β-sitostanol and campestanol as well as stanol esters were used. These characteristics allow both low-fat and high-fat foods to successfully incorporate PS and significantly lower LDL-c.

Topics: Anticholesteremic Agents; Cholesterol, LDL; Diet; Dietary Fats; Fatty Acids, Unsaturated; Functional Food; Humans; Hypercholesterolemia; Phytosterols; Phytotherapy; Plant Extracts; Sitosterols

Sitosterolemia is a rare, autosomal recessive inherited sterol storage disease associated with high tissue and serum plant sterol concentrations, caused by mutations in the adenosine triphosphate-bind-ing cassette (ABC) transporter ABCG5 or ABCG8 genes. Markedly increased serum concentration of plant sterols. such as sitosterol and campesterol, cause premature atherosclerosis and massive xanthomas. Hitherto known treatments for sitosterolemia, including a low-sterol diet, bile-salt binding resins, ileal bypass surgery and low density lipoprotein (LDL) apheresis have not yielded sufficient reduction of serum plant sterol levels and many patients show a sustained elevation of plant sterol levels, subsequently developing premature atherosclerotic cardiovascular diseases. Ezetimibe, an inhibitor of intestinal cholesterol absorption through its binding to Niemann-Pick C1-like 1 (NPC1L1), has been widely used for decreasing serum LDL-cholesterol levels in patients with hypercholesterolemia. Ezetimibe also reduces the gastrointestinal absorption of plant sterols, thereby also lowering the serum concentrations of plant sterols. This pharmacological property of ezetimibe shows its potential as a novel effective therapy for sitosterolemia. In the current review, we discuss the current therapy for patients with sitosterolemia and present two Japanese adolescent patients with this disease, one of whom underwent percutaneous coronary intervention for accelerated coronary atherosclerosis. Ezetimibe administration in addition to conventional drug therapy successfully reduced serum sitosterol levels by 51.3% and 48.9%, respectively, in the two patients, demonstrating ezetimibe as a novel and potent treatment agent for sitosterolemia that could work additively with conventional drug therapy.

Topics: Adolescent; Anticholesteremic Agents; ATP Binding Cassette Transporter, Subfamily G, Member 5; ATP-Binding Cassette Transporters; Azetidines; Bile Acids and Salts; Cardiovascular Diseases; Ezetimibe; Female; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Ileum; Ion Exchange Resins; Lipid Metabolism, Inborn Errors; Lipoproteins; Male; Models, Biological; Mutation, Missense; Phytosterols; Sitosterols; Young Adult

Normal human diet contains small amounts of phytosterols, mainly sitosterol and campesterol. Intestinal absorption of these plant sterols is low, about one tenth of that of cholesterol, such that their serum concentrations are also low, about 0.1 to 1% of the cholesterol levels. Like cholesterol they are transported by lipoproteins, mainly by LDL, and secreted unchanged in bile. Addition of plant sterols, or especially of their delta-5 saturated derivatives plant stanols into diet as fat-soluble esters inhibit cholesterol absorption and lower serum cholesterol similarly in short-term studies. Long-term consumption of plant stanol esters lowers serum cholesterol to the extent expected to reduce clinical manifestation of coronary heart disease by over 20% without detectable side effects, cholesterol lowering being especially effective in combination with cholesterol synthesis inhibitors statins.

Topics: Anticholesteremic Agents; Cholesterol; Diet; Humans; Intestinal Absorption; Lipids; Lipoproteins; Phytosterols; Plants, Edible; Sitosterols

Topics: Anticholesteremic Agents; Cholesterol; Cholesterol, LDL; Clinical Trials as Topic; Esters; Humans; Hypolipidemic Agents; Myocardial Ischemia; Phytosterols; Sitosterols

Functional foods enriched with plant sterols and stanols are on sale in many countries. Due to their structural similarity with cholesterol, these additives lower intestinal absorption of cholesterol, resulting in a 10-15% reduction in LDL-cholesterol when their daily intakes are 2-3 g. They are also effective as part of a cholesterol-lowering diet and in combination with cholesterol-lowering drugs. Estimates for the absorption of plant sterols (sitosterol and campesterol) and of campestanol are around 10%, and for sitostanol less than 5%. Lipid-standardized plasma levels are very low, but increase when statins are used. Extensive toxicological evaluation studies have not revealed any harmful side-effects. In human studies, side-effects were comparable to placebo treatment. However, lipid-standardized levels of the hydrocarbon carotenoids may decrease, without leaving the normal range. Together, these findings indicate that these functional foods have great potential in the prevention of coronary heart disease. However, post-marketing surveillance for example for functional foods in general is necessary to monitor possible adverse effects and describe consumers and consumption patterns.

Topics: Anticholesteremic Agents; Cardiovascular Diseases; Cholesterol; Diet; Food, Organic; Humans; Intestinal Absorption; Lipid Metabolism; Phytosterols; Risk Factors; Safety; Sitosterols

To assess the association between biomarkers of thyroid status and 5α-stanols in patients with sitosterolemia treated with ezetimibe (EZE).. Eight patients with sitosterolemia (16-56 years of age) were studied during 14 weeks off EZE therapy and 14 weeks on EZE (10 mg/day). Serum thyroid biomarkers (free triiodothyronine [FT3], free thyroxine [FT4], FT3/FT4 ratio, thyroid-stimulating hormone), 5α-stanols (sitostanol and cholestanol), and cholestanol precursors (total cholesterol and its synthesis marker lathosterol, and 7α-hydroxy-4-cholesten-3-one cholestenol) were measured at baseline and during the 14 weeks off EZE and on EZE.. EZE increased FT3/FT4 (10% ± 4%; P = .02). EZE reduced plasma and red blood cells sitostanol (-38% ± 6% and -20% ± 4%; all P < .05) and cholestanol (-18% ± 6% and -13% ± 3%; all P < .05). The change in plasma cholestanol level on EZE inversely correlated with the change in FT3/FT4 (r = -0.86; P = .01). EZE lowered total cholesterol (P < .0001) and did not affect 7α-hydroxy-4-cholesten-3-one cholestanol. EZE increased (P < .0001) lathosterol initially, but the level was not sustained, resulting in similar levels at week 14 off EZE and on EZE.. In patients with STSL, 5α-stanols levels might be associated with thyroid function. EZE reduces circulating 5α-stanols while increasing FT3/FT4, implying increased conversion of T4 to T3, thus possibly improving thyroid hormone status.. ClinicalTrials.govNCT01584206.

Topics: Adolescent; Adult; Anticholesteremic Agents; Cholestanol; Cholestenones; Cholesterol; Ezetimibe; Female; Humans; Hypercholesterolemia; Intestinal Diseases; Lipid Metabolism, Inborn Errors; Male; Middle Aged; Phytosterols; Sitosterols; Thyrotropin; Thyroxine; Triiodothyronine; Young Adult

Consumption of plant stanols and plant sterols decreases LDL cholesterol level and increases serum concentrations of plant stanols/sterols, but it is practically unexplored whether also their tissue concentrations increase. Thus, the aim of this study was to assess whether consuming plant stanols/sterols increases their concentrations in stenotic aortic valves and affect the valvular structure (collagen and elastin) or inflammation (macrophages and mast cells).. In a randomized, double-blind controlled intervention patients with severe aortic stenosis consumed margarine without (n = 11) or with 2 g of plant stanols (n = 12) or sterols (n = 13) until valve replacement surgery (2.6 months, on average). The effects of sitostanol and sitosterol on the expression and secretion of proinflammatory cytokines by cultured aortic valve myofibroblasts were also assessed.. Control-related LDL-cholesterol was diminished by 16% (p < 0.05) by plant stanol and by 11% (NS) by plant sterol consumption, respectively. In the resected valves, cholesterol, plant stanol and sterol levels were similar in all groups. Consumed plant stanols or sterols had no effect on valvular structure or mast cell or macrophage numbers in valves. Incubation of cultured myofibroblasts derived from stenotic valves with sitostanol or sitosterol decreased mRNA expression of the monocyte chemotactic protein-1 (p < 0.05) and interleukin-1 beta (p < 0.05).. In this study, plant stanol/sterol consumption did not affect cholesterol, plant stanol or sterol levels in stenotic aortic valves; neither did they influence the structure or the inflammatory status of the valves. However, these findings need to be confirmed in a larger-scale intervention. ClinicalTrials.govRegister #NCT00738933.

Topics: Aged; Aged, 80 and over; Aortic Valve; Chemokine CCL2; Cholesterol, LDL; Diet; Double-Blind Method; Female; Heart Valve Prosthesis Implantation; Humans; Interleukin-1beta; Male; Margarine; Middle Aged; Myofibroblasts; Phytosterols; RNA, Messenger; Sitosterols

Clinical safety of consuming plant stanol ester spreads during pregnancy and lactation, the impact on maternal and infant serum and breast-milk cholesterol and the ratios (micromol/mmol of cholesterol) of synthesis and absorption markers were evaluated. Pregnant women (n 21) were randomised to control and dietary intervention groups, the intervention including advice to follow a balanced diet and to consume spreads enriched with plant stanol esters. Participants were followed during and after pregnancy and their infants up to 1 year of age. A mean 1.1 (sd 0.4) g consumption of plant stanols during pregnancy and 1.4 (sd 0.9) g 1 month post-partum increased sitostanol and the markers for cholesterol synthesis, lathosterol, lathosterol/campesterol and lathosterol/sitosterol, and reduced a marker for cholesterol absorption, campesterol, in maternal serum. In breast milk, desmosterol was lower in the intervention group, while no differences were detected between the groups in infants' serum. Plant stanol ester spread consumption had no impact on the length of gestation, infants' growth or serum beta-carotene concentration at 1 and 6 months of age, but the cholesterol-adjusted serum beta-carotene concentration was lowered at 1 month in the intervention group. Plant stanol ester spread consumption appeared safe in the clinical setting, except for potential lowering of infants' serum beta-carotene concentration, and was reflected in the markers of cholesterol synthesis and absorption in mothers' serum, encouraging further studies in larger settings.

Topics: Analysis of Variance; beta Carotene; Biomarkers; Child Development; Cholesterol; Desmosterol; Female; Humans; Infant; Infant, Newborn; Lactation; Margarine; Milk, Human; Phytosterols; Pregnancy; Safety; Sitosterols; Squalene

Consumption of plant sterol- or stanol-enriched margarines by statin users results in an additional LDL-cholesterol reduction of approximately 10 %, which may be larger than the average decrease of 3-7 % achieved by doubling the statin dose. However, whether this effect persists in the long term is not known. Therefore, we examined in patients already on stable statin treatment the effects of 85 weeks of plant sterol and stanol ester consumption on the serum lipoprotein profile, cholesterol metabolism, and bile acid synthesis. For this, a double-blind randomised trial was designed in which fifty-four patients consumed a control margarine with no added plant sterols or stanols for 5 weeks (run-in period). For the next 85 weeks, seventeen subjects continued with the control margarine and the other two groups with either a plant sterol (n 18) or plant stanol (n 19) (2.5 g/d each) ester-enriched margarine. Blood was sampled at the end of the run-in period and every 20 weeks during the intervention period. Compared with the control group, plant sterol and stanol ester consumption reduced LDL-cholesterol by 0.28 mmol/l (or 8.7 %; P = 0.08) and 0.42 mmol/l (13.1 %; P = 0.006) respectively after 85 weeks. No effects were found on plasma concentrations of oxysterols or 7 alpha-hydroxy-4-cholesten-3-one, a bile acid synthesis marker. We conclude that long-term consumption of both plant sterol and stanol esters effectively lowered LDL-cholesterol concentrations in statin users.

Topics: Analysis of Variance; Anticholesteremic Agents; Biomarkers; Cholestenones; Cholesterol; Cholesterol, LDL; Double-Blind Method; Esters; Female; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypercholesterolemia; Lipid Metabolism; Lipoproteins; Male; Margarine; Middle Aged; Phytosterols; Sitosterols; Stigmasterol

Plant sterol/stanol margarines are recommended as a lipid-lowering dietary supplement in the treatment of hypercholesterolemia. Parameters predicting the individual cholesterol-lowering effect have not been elucidated so far. Therefore, we investigated the responsiveness to sitostanol-supplemented margarine in a specially selected population.. From a total number of 137 male subjects with hypercholesterolemia, eight subjects with the lowest and eight subjects with the highest ratios of lathosterol to campesterol in serum were included in the study. They received 1 g sitostanol-supplemented margarine b.i.d. for four weeks. Serum lipoproteins, the cholesterol precursor lathosterol, the plant sterols campesterol and sitosterol were measured. Subjects with a low ratio of lathosterol to campesterol had a significant decrease of serum total cholesterol (-14.2%; p < 0.01) and LDL cholesterol (-13.8%; p < 0.01; responder). In subjects with a high ratio there was no significant change in total cholesterol and LDL cholesterol (2.2 and 4.3%; non-responder).. The ratio of serum lathosterol to campesterol predicts the reduction of total cholesterol and LDL cholesterol during administration of sitostanol-supplemented margarine in patients with mild hypercholesterolemia.

Topics: Adult; Anticholesteremic Agents; Cholesterol; Cholesterol, Dietary; Cholesterol, HDL; Cholesterol, LDL; Humans; Hypercholesterolemia; Male; Margarine; Middle Aged; Patient Selection; Phytosterols; Predictive Value of Tests; Sitosterols; Triglycerides

Feeding of margarines containing sitostanol (CAS 19466-47-8), sitostanol acetate (CAS 73052-08-1), sitostanol oleate (CAS 107615-79-2), or placebo (equivalent of 0.5 g of sitostanol t.i.d) on cholesterol absorption and serum lipids were studied in 10 normolipemic volunteers in a randomized double blind cross-over trial. The study was divided into an open one-week run-in phase and four one-week treatment periods. Each treatment week was followed by a two-week washout period. Measurements of cholesterol absorption was performed by the continuous isotope feeding method using stable isotope labeled cholesterol and sitostanol. Cholesterol absorption during placebo, sitostanol, sitostanol acetate and sitostanol oleate feeding averaged 41.6 +/- (SD) 8.0, 10.2 +/- 6.6, 17.0 +/- 6.7, and 20.5 +/- 5.3%, respectively (p < 0.001 for all against placebo). Low density lipoprotein (LDL) cholesterol was proportionally reduced by 22 (p < 0.001), 14 (p < 0.05), and 8% (ns). Absorption efficiency was significantly lower with free sitostanol than with sitostanol acetate or oleate (p < 0.01). Percent reduction in cholesterol absorption with all preparations compared to placebo correlated positively with the percent reduction in LDL cholesterol (r = 0.404; p < 0.03). The results indicate that unesterified sitostanol is more effective in inhibiting cholesterol absorption and reducing LDL cholesterol than the acetate or oleate esters.

Topics: Acetates; Adult; Cholesterol, Dietary; Cross-Over Studies; Double-Blind Method; Humans; Intestinal Absorption; Lipoproteins; Male; Margarine; Oleic Acids; Phytosterols; Sitosterols

In a randomized, double-blind, placebo-controlled trial we evaluated the effect of dietary chocolates enriched with a wood-based phytosterol-phytostanol mixture, containing 18 % (w/w) sitostanol, compared with placebo dietary chocolates in seventy subjects with primary hypercholesterolaemia (total cholesterol levels below 8 mmol/l). For 4 weeks, participants consumed three servings of the phytosterol-enriched chocolate/d that provided 1.8 g unesterified phytosterols/d or a placebo chocolate in conjunction with a low-fat, low-cholesterol diet. Plasma total and LDL-cholesterol levels were statistically significantly reduced by 6.4 % (-0.44 mmol/l) and 10.3 % (-0.49 mmol/l), respectively, after 4 weeks of phytosterol-enriched-chocolate treatment. Plasma HDL-cholesterol and triacylglycerol levels were not affected. Consumption of phytosterol-enriched chocolates significantly increased plasma lathosterol concentration (+20.7 %), reflecting an increased endogenous cholesterol synthesis in response to phytosterol-induced decreased intestinal cholesterol absorption. Furthermore, the chocolates enriched with phytosterols significantly increased both plasma sitosterol (+95.8 %) and campesterol (+64.1 %) levels, compared with the placebo chocolate group. However, the absolute values of plasma sitosterol and campesterol remained within the normal range, that is, below 10 mg/l. The chocolates with phytosterols were palatable and induced no clinical or biochemical side effects. These findings indicate that dietary chocolate enriched with tall oil-derived phytosterols (1.8 g/d) is effective in lowering blood total and LDL-cholesterol levels in subjects with mild hypercholesterolaemia and thus may be helpful in reducing the risk of CHD in these individuals.

Topics: Adult; Apolipoproteins B; Cacao; Chi-Square Distribution; Cholesterol; Cholesterol, LDL; Double-Blind Method; Female; Humans; Hypercholesterolemia; Lipids; Male; Middle Aged; Phytosterols; Plant Oils; Sitosterols; Statistics, Nonparametric

Plant stanols lower intestinal cholesterol absorption. This causes a decrease in serum low-density lipoprotein (LDL)-cholesterol, despite a compensatory increase in cholesterol synthesis. We therefore hypothesized that plant stanols also change LDL-cholesterol-standardized concentrations of ubiquinol-10 (a side product of the cholesterol synthesis cascade) and of those fat-soluble antioxidants that are mainly carried by LDL. To examine this, 112 nonhypercholesterolemic subjects consumed low erucic acid rapeseed oil (LEAR)-based margarine and shortening for 4 weeks. For the next 8 weeks, 42 subjects consumed the same products, while the other subjects received products with vegetable oil-based stanols (2.6 g sitostanol plus 1.2 g campestanol daily, n = 36) or wood-based stanols (3.7 g sitostanol plus 0.3 g campestanol daily, n = 34). Consumption of both plant stanol ester mixtures increased cholesterol synthesis and lowered cholesterol absorption, as indicated by increased serum cholesterol-standardized lathosterol and decreased plant sterol concentrations, respectively. Compared with the control group, absolute plasma ubiquinol-10 concentrations were lowered by 12.3% +/- 18.9% (-0.14 microg/mL v. the control group; P =.004; 95% confidence interval [CI] for the difference in changes, -0.05 to -0.22 microg/mL) in the vegetable oil-based group and by 15.4% +/- 13.0% (-0.17 microg/mL v. the control group; P <.001; 95% CI for the difference, -0.08 to -0.27 microg/mL) in the wood-based group. Changes in LDL-cholesterol-standardized ubiquinol-10 concentrations were not significantly changed. The most lipophylic antioxidants, the hydrocarbon carotenoids (beta-carotene, alpha-carotene, and lycopene), decreased most, followed by the less lipophylic oxygenated carotenoids (lutein/zeaxanthin and beta-cryptoxanthin) and the tocopherols. These reductions were related to the reduction in LDL, which carry most of these antioxidants. The decrease in the hydrocarbon carotenoids, however, was also significantly associated with a decrease in cholesterol absorption. LDL-cholesterol-standardized antioxidant concentrations were not changed, except for beta-carotene, which was still, although not significantly, lowered by about 10%. We conclude that the increase in endogenous cholesterol synthesis during plant stanol ester consumption does not result in increased LDL-cholesterol-standardized concentrations of ubiquinol-10, a side product of the cholesterol synthesis cascade. Fur

Topics: Absorption; Adolescent; Adult; Antioxidants; Carotenoids; Cholesterol; Cholesterol, LDL; Diet; Erucic Acids; Fats; Fatty Acids, Monounsaturated; Female; Humans; Male; Margarine; Middle Aged; Phytosterols; Plant Oils; Plants, Edible; Rapeseed Oil; Sitosterols; Solubility; Ubiquinone; Vitamin A; Vitamin E; Wood

A pine wood based stanol ester mixture-composed of sitostanol (92%) and campestanol (8%) effectively lowers cholesterol absorption and consequently LDL-cholesterol concentrations. It has been postulated that the less absorbable plant sterols reduce cholesterol absorption more effectively. As sitostanol is absorbed less than campestanol, we decided to examine if a vegetable oil based stanol ester mixture with 68% sitostanol and 32% campestanol is less effective than the wood based stanol ester mixture. For this, 112 non-hypercholesterolemic men and women consumed for 4 weeks a rapeseed oil (LEAR) based margarine and shortening. For the next 8 weeks, 42 subjects continued with these products, while the other subjects received products with a vegetable oil (n=36) or a pine wood based stanol ester mixture (n=34). Consumption of 3.8 g vegetable oil based stanols (2.6 g sitostanol plus 1.2 g campestanol) lowered LDL cholesterol 14.6+/-8.0% (-0.37 mmol/l; vs. the control group; P<0.001; 95% CI for the difference, -0.22 to -0. 51 mmol/l). Four grams pine wood based stanols (3.7 g sitostanol plus 0.3 g campestanol) showed a comparable decrease of 12.8+/-11.2% (-0.34 mmol/l; P<0.001; 95% CI-0.18 to-0.51 mmol/l). Decreases in LDL cholesterol were not different between the two experimental groups (P=0.793), while apoE genotype did not have a major impact on this hypocholesterolemic response. Serum HDL cholesterol and triacylglycerol concentrations were not changed. The decreases in apo B in both experimental groups differed significantly (P<0.001) from changes in the control group. Coagulation and fibrinolytic parameters were not affected. We therefore conclude that vegetable oil and wood based stanol ester mixtures, with a different sitostanol/campestanol ratio, have similar LDL cholesterol lowering effects in a non-hypercholesterolemic population.

Topics: Adult; Anticholesteremic Agents; Apolipoproteins E; Blood Coagulation; Body Weight; Cholesterol; Drug Combinations; Fatty Acids, Monounsaturated; Female; Fibrinolysis; Hemostasis; Humans; Lipids; Male; Middle Aged; Phytosterols; Plant Oils; Polymorphism, Genetic; Rapeseed Oil; Reference Values; Sitosterols

Plant sterols are natural dietary components with serum cholesterol-lowering properties. The lowering of serum cholesterol by plant sterols is believed to be the result of an inhibition of cholesterol absorption in the small bowel, although increased bile acid excretion has also been suggested. The difference in effect of saturated and unsaturated plant sterols on cholesterol absorption needs to be elucidated further.. The primary aim of this study was to measure small-bowel cholesterol absorption and sterol excretion in addition to hepatic cholesterol synthesis after intake of soy sterol esters and beta-sitostanol ester corresponding to 1.5 g plant sterols/d.. Seven ileostomy subjects were studied during a control period and 2 intervention periods when either soy sterol esters or beta-sitostanol ester was added to a basal diet. Ileostomy bags were collected every other hour and frozen immediately for analysis of nutrients and sterols.. Cholesterol absorption was 56% (43-65%) in the control period and decreased to 38% (32-46%) in the soy sterol ester period (P = 0.00) and to 39% (30-48%) in the beta-sitostanol ester period (P = 0.00).. Esterified soy sterols and beta-sitostanol inhibited cholesterol absorption equally, despite the different structures of the plant sterols.

Topics: Adult; Aged; Anticholesteremic Agents; Cholesterol; Colitis, Ulcerative; Esters; Female; Glycine max; Humans; Ileostomy; Intestinal Absorption; Intestine, Small; Liver; Male; Middle Aged; Phytosterols; Sitosterols; Sterols

The effect of plant stanol ester on serum cholesterol is dose-dependent. However, it is not clear what the dose is beyond which no additional benefit can be obtained. Therefore, we determined the dose-response relationship for serum cholesterol with different doses of plant stanol ester in hypercholesterolemic subjects. In a single-blind design each of 22 men or women consumed five different doses of plant stanol [target (actual) intake 0 (0), 0.8 (0.8), 1.6 (1.6), 2.4 (2.3), 3.2 (3.0) g/d] added as plant stanol esters to margarine for 4 wk. The order of dose periods was randomly determined. Serum total cholesterol concentration decreased (calculated in reference to control) by 2.8% (P = 0.384), 6.8% (P < 0.001), 10.3% (P < 0.001) and 11.3% (P < 0.001) by doses from 0.8 to 3.2 g. The respective decreases for LDL cholesterol were 1.7% (P = 0. 892), 5.6% (P < 0.05), 9.7% (P < 0.001) and 10.4% (P < 0.001). Although the decreases were numerically greater with 2.4 and 3.2 g doses than with the 1.6 g dose, these differences were not significant (P = 0.054-0.516). Serum plant stanols rose slightly, but significantly with the dose (P < 0.001). Apolipoprotein B concentration was decreased significantly already at the dose of 0.8 g (8.7%, P < 0.001). Apolipoprotein E genotype did not affect the lipid responses. We conclude that significant reduction of serum total and LDL cholesterol concentrations is reached with the 1.6-g stanol dose, and increasing the dose from 2.4 to 3.2 g does not provide clinically important additional effect.

Topics: Adult; Aged; Anticholesteremic Agents; Carotenoids; Child, Preschool; Cholesterol; Dose-Response Relationship, Drug; Esters; Female; Humans; Hypercholesterolemia; Lipids; Lipoproteins; Male; Margarine; Middle Aged; Osmolar Concentration; Phytosterols; Single-Blind Method; Sitosterols; Vitamins

To examine in humans the effects on serum lipids, lipoproteins and fat-soluble antioxidants of a daily consumption of 2.5 g plant stanols, consumed either once per day at lunch or divided over the three meals.. A randomized, double-blind, placebo-controlled, cross-over design.. Thirty-nine healthy normocholesterolemic or mildly hypercholesterolemic subjects participated.. Each subject consumed in random order; no plant stanols; 2.5 g plant stanols at lunch; and 2.5 g plant stanols divided over the three meals (0.42 g at breakfast, 0.84 g at lunch and 1.25 g at dinner, which is proportional to dietary cholesterol intake). Each period lasted 4 weeks. Plant stanols were esterified with fatty acids from low erucic rapeseed oil (LEAR) and incorporated into margarines or shortenings.. Consumption of 2.5 g plant stanols at lunch results in a similar low-density lipoprotein (LDL)-cholesterol-lowering efficacy compared to consumption of 2.5 g plant stanols divided over the three meals (-0. 29 mmol/l compared with the control period (P<0.001; 95% CI, -0.19 to -0.39 mmol/l) for the once per day diet and -0.31 mmol/l (P<0. 001; 95% CI, -0.20 to -0.41 mmol/l)) for the three times per day period). High-density Lipoprotein (HDL) cholesterol and triacylglycerol concentrations did not change. After standardization for LDL cholesterol, the sum of the most lipophylic hydrocarbon carotenoids (ie alpha-carotene, beta-carotene and lycopene) in particular was slightly, though not significantly, lowered by -0. 017+/-0.018 micromol/mmol LDL cholesterol (P=0.307) after the once per day period and by -0.032+/-0.016 micromol/mmol LDL cholesterol (P=0.049) after the three times per day period.. Our findings suggest that for lowering LDL cholesterol concentrations it is not necessary to consume products rich in plant stanol ester at each meal or simultaneously with dietary cholesterol.. Raisio Group, Raisio, Finland.

Topics: Adolescent; Adult; Anticholesteremic Agents; Antioxidants; Cholesterol, LDL; Cross-Over Studies; Dietary Fats; Double-Blind Method; Esters; Female; Humans; Intestinal Mucosa; Lipids; Lipoproteins; Male; Margarine; Middle Aged; Netherlands; Phytosterols; Plants; Sitosterols; Surveys and Questionnaires; Time Factors

To investigate cholesterol-lowering effects of stanol ester (STAEST) and sterol ester (STEEST)-enriched margarines as part of a low-fat diet.. According to a Latin square model randomized double-blind repeated measures design with three test margarines and three periods.. Outpatient clinical trial with free-living subjects.. Thirty-four hypercholesterolaemic subjects completed the study.. Subjects consumed three rapeseed oil-based test margarines (STAEST, STEEST and control (no added stanols or sterols)) as part of a low-fat diet each for 4 weeks.. Mean daily intake of total plant sterols plus stanols was 2.01-2.04 g during the two test margarine periods. In reference to control, serum total cholesterol was reduced by 9.2 and 7.3% with the STAEST and STEEST margarine, respectively (P<0.001 for both). The respective reductions for low-density lipoprotein (LDL) cholesterol were 12.7 and 10.4% (P<0. 001). The cholesterol-lowering effects of the test margarines did not differ significantly. The presence of apolipoprotein E4 allele had a significant effect on LDL cholesterol response during the STAEST margarine only. Serum sitosterol and campesterol increased by 0.83 and 2.77 mg/l with the STEEST (P<0.001), respectively and decreased by 1.18 and 2.60 mg/l with the STAEST margarine (P<0.001). Increases of serum sitostanol and campestanol were 0.11 and 0.19 mg/l with the STAEST margarine (P<0.001), repsectively. No significant changes were found in serum fat-soluble vitamin and carotenoid concentrations when related to serum total cholesterol.. STAEST and STEEST margarines reduced significantly and equally serum total and LDL cholesterol concentrations as part of a low-fat diet.. Grant to the University of Kuopio by Raisio Benecol Ltd, Raisio, Finland.

Topics: Adult; Aged; Anticholesteremic Agents; Antioxidants; Carotenoids; Cholesterol; Diet, Fat-Restricted; Double-Blind Method; Esters; Female; Humans; Hypercholesterolemia; Male; Margarine; Middle Aged; Phytosterols; Plant Oils; Sitosterols; Vitamin E

We investigated the changes of cholesterol and non-cholesterol sterol metabolism during plant stanol ester margarine feeding in 153 hypercholesterolemic subjects. Rapeseed oil (canola oil) margarine without (n = 51) and with (n = 102) stanol (2 or 3 g/day) ester was used for 1 year. Serum sterols were analyzed with gas-liquid chromatography. The latter showed a small increase in sitostanol peak during stanol ester margarine eating. Cholestanol, campesterol, and sitosterol proportions to cholesterol were significantly reduced by 5-39% (P < 0.05 or less for all) by stanol esters; the higher their baseline proportions the higher were their reductions. The precursor sterol proportions were significantly increased by 10- 46%, and their high baseline levels predicted low reduction of serum cholesterol. The decrease of the scheduled stanol dose from 3 to 2 g/day after 6-month feeding increased serum cholesterol by 5% (P < 0. 001) and serum plant sterol proportions by 8-13% (P < 0.001), but had no consistent effect on precursor sterols. In twelve subjects, the 12-month level of LDL cholesterol exceeded that of baseline; the non-cholesterol sterol proportions suggested that stimulated synthesis with relatively weak absorption inhibition contributed to the non-responsiveness of these subjects. In conclusion, plant stanol ester feeding lowers serum cholesterol in about 88% of subjects, decreases the non-cholesterol sterols that reflect cholesterol absorption, increases the sterols that reflect cholesterol synthesis, but also slightly increases serum plant stanols. Low synthesis and high absorption efficiency of cholesterol results in the greatest benefit from stanol ester consumption.

Topics: Anticholesteremic Agents; Body Mass Index; Cholestanol; Cholesterol; Dietary Fats, Unsaturated; Esters; Fatty Acids, Monounsaturated; Humans; Hypercholesterolemia; Kinetics; Margarine; Phytosterols; Rapeseed Oil; Sitosterols; Sterols

Our aim was to investigate (1) whether different campestanol/sitostanol mixtures in margarine differ in reducing serum cholesterol, and (2) whether sitostanol ester in butter decreases serum cholesterol and alters cholesterol absorption and metabolism. Twenty-three postmenopausal women replaced 25 g dietary fat with (1) sitostanol ester-rich (campestanol to sitostanol ratio 1:11) and (2) campestanol ester-rich (campestanol to sitostanol ratio 1:2) rapeseed oil margarine, (3) butter, and (4) sitostanol ester-rich (campestanol to sitostanol ratio 1:13) butter. The respective scheduled stanol intake was 3.18, 3.16, and 2.43 g/d. The 6-week margarine periods and, after an 8-week washout, 5-week butter periods were double-blind and in random order. Serum cholesterol precursor sterols (indicators of cholesterol synthesis) and plant sterols (indicators of cholesterol absorption) were quantified with gas-liquid chromatography (GLC). Low-density lipoprotein (LDL) cholesterol was reduced by 8% and 10% with the sitostanol and campestanol ester-rich margarines versus baseline (P < .05 for both) and high-density lipoprotein (HDL) cholesterol was increased by 6% and 5% (P < .05), so the LDL/HDL cholesterol ratio was reduced by 15% (P < .05 for both). Sitostanol ester-rich butter decreased LDL cholesterol 12% and the LDL/HDL cholesterol ratio 11% (P < .05 for both) versus the butter period. The serum proportions of plant sterols and cholestanol were similarly reduced and those of cholesterol precursor sterols were similarly increased during all periods (P < .05 for all). Serum proportions of sitostanol and campestanol were slightly increased, indicating that their absorption related to their dietary intake. During all stanol interventions, serum vitamin D and retinol concentrations and alpha-tocopherol to cholesterol ratios were unchanged, whereas those of alpha- and beta-carotenes were significantly reduced. We conclude that varying the campestanol to sitostanol ratio from 1:13 to 1:2 in margarine and in butter similarly decreased cholesterol absorption, LDL cholesterol, and the LDL/HDL cholesterol ratio such that the serum lipids became less atherogenic.

Topics: Absorption; Anticholesteremic Agents; Butter; Cholesterol; Cohort Studies; Dietary Fats; Double-Blind Method; Drug Combinations; Fatty Acids, Monounsaturated; Female; Humans; Male; Margarine; Middle Aged; Phytosterols; Plant Extracts; Plant Oils; Rapeseed Oil; Sitosterols; Sterols

Dietary plant sterols (phytosterols) have been shown to lower plasma lipid concentrations in animals and humans. However, the effect of phytosterol intake from tall oil on cholesterol and phytosterol metabolism has not been assessed in subjects fed precisely controlled diets.. Our objective was to examine the effects of sitostanol-containing phytosterols on plasma lipid and phytosterol concentrations and de novo cholesterol synthesis rate in the context of a controlled diet.. Thirty-two hypercholesterolemic men were fed either a diet of prepared foods alone or a diet containing 1.7 g phytosterols/d for 30 d in a parallel study design.. No overall effects of diet on total cholesterol concentrations were observed, although concentrations were lower with the phytosterol-enriched than with the control diet on day 30 (P < 0.05). LDL-cholesterol concentrations on day 30 had decreased by 8.9% (P < 0.01) and 24.4% (P < 0.001) with the control and phytosterol-enriched diets, respectively. HDL-cholesterol and triacylglycerol concentrations did not change significantly. Moreover, changes in circulating campesterol and beta-sitosterol concentrations were not significantly different between phytosterol-fed and control subjects. In addition, there were no significant differences in fractional (0.091 +/- 0.028 and 0.091 +/- 0.026 pool/d, respectively) or absolute (0.61 +/- 0.24 and 0.65 +/- 0.23 g/d, respectively) synthesis rates of cholesterol observed between control and phytosterol-fed subjects.. Addition of blended phytosterols to a prudent North American diet improved plasma LDL-cholesterol concentrations by mechanisms that did not result in significant changes in endogenous cholesterol synthesis in hypercholesterolemic men.

Topics: Adult; Anticholesteremic Agents; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Chromatography, Gas; Humans; Hyperlipidemias; Male; Middle Aged; Phytosterols; Sitosterols; Triglycerides

To determine the efficacy of stanol esters in lowering cholesterol in a US population.. After a run-in phase, 318 subjects were randomized to receive one of the following margarine-like spreads containing stanol ester or placebo for 8 weeks: EU 3 G: 1 g of stanol (ester form) per 8-g serving of a European formula 3 times a day; US 3 G: 1 g of stanol (ester form) per 8-g serving of a US reformulation 3 times a day; US 2 G: 0.67 g of stanol (ester form) per 8-g serving of a US reformulation 3 times a day; or placebo spread.. Mean +/- SD baseline total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) levels were 233+/-20 and 153+21 mg+/-dL, respectively. In the US 3 G group, 3 g daily of stanol esters lowered TC and LDL-C levels by 6.4% and 10.1%, respectively. There was a dose-dependent response compared with 2 g daily (US 2 G). Triglyceride and high-density lipoprotein cholesterol levels were unchanged. The incidence of adverse effects was not different from placebo. Serum vitamin A and 25-hydroxyvitamin D levels were not affected.. Stanol esters lowered TC and LDL-C levels in a mildly hypercholesterolemic US population without evidence of adverse effects. It may be a useful dietary adjunct to lower cholesterol.

Topics: Adult; Anticholesteremic Agents; beta Carotene; Cholestanols; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Dietary Fats; Dose-Response Relationship, Drug; Double-Blind Method; Esters; Female; Humans; Hypercholesterolemia; Male; Middle Aged; Phytosterols; Sitosterols; Treatment Outcome; Triglycerides; United States; Vitamin A; Vitamin D

To compare effects on plasma total-, LDL-, and HDL-cholesterol concentrations of margarines enriched with different vegetable oil sterols or sitostanol-ester.. A randomized double-blind placebo-controlled balanced incomplete Latin square design with five treatments and four periods of 3.5 weeks. Margarines enriched with sterols from soybean, sheanut or ricebran oil or with sitostanol-ester were compared to a non-enriched control margarine. Sterol intake was between 1.5-3.3 g/d. Two thirds of the soybean oil sterols were esterified to fatty acids.. Unilever Research Laboratory, Vlaardingen, The Netherlands.. One hundred healthy non-obese normocholesterolaemic and mildly hypercholesterolaemic volunteers aged 45+/-12.8 y, with plasma total cholesterol levels below 8 mmol/L at entry.. Plasma lipid, carotenoid and sterol concentrations, blood clinical chemistry and haematology, fatty acid composition of plasma cholesterylesters and food intake.. Ninety-five volunteers completed the study. None of the margarines induced adverse changes in blood clinical chemistry, serum total bile acids or haematology. Plasma total- and LDL-cholesterol concentrations were significantly reduced by 8-13% (0.37-0.44 mmol/L) compared to control for margarines enriched in soybean oil sterol-esters or sitostanol-ester. No effect on HDL-cholesterol concentrations occurred. The LDL- to HDL-cholesterol ratio was reduced by 0.37 and 0.33 units for these margarines, respectively. Effects on blood lipids did not differ between normocholesterolaemic and mildly hypercholesterolaemic subjects. Plasma sitosterol and campesterol levels were significantly higher for the soybean oil sterol margarine and significantly lower for the sitostanol-ester margarine compared to control. Dietary intake was very similar across treatments. The fatty acid composition of plasma cholesterylesters confirmed the good compliance to the treatment. All sterol enriched margarines reduced lipid-standardized plasma alpha- plus beta-carotene levels. Plasma lycopene levels were also reduced but this effect was not significant for all products.. A margarine with sterol-esters from soybean oil, mainly esters from sitosterol, campesterol and stigmasterol, is as effective as a margarine with sitostanol-ester in lowering blood total- and LDL-cholesterol levels without affecting HDL-cholesterol concentrations. Incorporation in edible fat containing products of such substances may substantially reduce the risk of cardiovascular disease in the population.

Topics: Adult; Carotenoids; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Dietary Fats, Unsaturated; Double-Blind Method; Humans; Hypercholesterolemia; Margarine; Middle Aged; Phytosterols; Placebos; Plant Oils; Sitosterols; Soybean Oil

Dietary plant sterols, especially sitostanol, reduce serum cholesterol by inhibiting cholesterol absorption. Soluble sitostanol may be more effective than a less soluble preparation. We tested the tolerability and cholesterol-lowering effect of margarine containing sitostanol ester in a population with mild hypercholesterolemia.. We conducted a one-year, randomized, double-blind study in 153 randomly selected subjects with mild hypercholesterolemia. Fifty-one consumed margarine without sitostanol ester (the control group), and 102 consumed margarine containing sitostanol ester (1.8 or 2.6 g of sitostanol per day).. The margarine containing sitostanol ester was well tolerated. The mean one-year reduction in serum cholesterol was 10.2 percent in the sitostanol group, as compared with an increase of 0.1 percent in the control group. The difference in the change in serum cholesterol concentration between the two groups was -24 mg per deciliter (95 percent confidence interval, -17 to -32; P < 0.001). The respective reductions in low-density lipoprotein (LDL) cholesterol were 14.1 percent in the sitostanol group and 1.1 percent in the control group. The difference in the change in LDL cholesterol concentration between the two groups was -21 mg per deciliter (95 percent confidence interval, -14 to -29; P < 0.001). Neither serum triglyceride nor high-density lipoprotein cholesterol concentrations were affected by sitostanol. Serum campesterol, a dietary plant sterol whose levels reflect cholesterol absorption, was decreased by 36 percent in the sitostanol group, and the reduction was directly correlated with the reduction in total cholesterol (r = 0.57, P < 0.001).. Substituting sitostanol-ester margarine for part of the daily fat intake in subjects with mild hypercholesterolemia was effective in lowering serum total cholesterol and LDL cholesterol.

Topics: Adult; Anticholesteremic Agents; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Dietary Fats; Double-Blind Method; Female; Humans; Hypercholesterolemia; Male; Margarine; Middle Aged; Phytosterols; Sitosterols; Triglycerides

Topics: Cholesterol; Chromatography, High Pressure Liquid; Flow Injection Analysis; Ion Mobility Spectrometry; Limit of Detection; Mass Spectrometry; Phytosterols; Sitosterols; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Lowering of LDL cholesterol levels by plant sterols and stanols is associated with decreased risk of cardiovascular disease in humans. Plant sterols and stanols also lower triacylglycerol (TG). However, it is not fully understood how reduction in TG is achieved and what the full potential of plant sterols and stanols is on whole-body metabolism. We here hypothesize that high levels of plant sterols and stanols stimulate whole-body energy expenditure, which can be attributed to changes in mitochondrial function of brown adipose tissue (BAT), skeletal muscle and liver.. Phytosterolemic mice were fed chow diets for 32 weeks to examine whole-body weight gain. In vitro, 24-h incubation were performed in adipocytes derived from human BAT, human myotubes or HepG2 human hepatocytes using sitosterol or sitostanol. Following mitochondrial function was assessed using seahorse bioanalyzer.. Chow feeding in phytosterolemic mice resulted in diminished increase in body weight compared to control mice. In vitro, sitosterol or sitostanol did not change mitochondrial function in adipocytes derived from human BAT or in cultured human myotubes. Interestingly, maximal mitochondrial function in HepG2 human hepatocytes was decreased following sitosterol or sitostanol incubation, however, only when mitochondrial function was assessed in low glucose-containing medium.. Beneficial in vivo effects of plant sterols and stanols on lipid and lipoprotein metabolism are well recognized. Our results indicate that alterations in human mitochondrial function are apparently not involved to explain these beneficial effects.

Topics: Adipocytes, Brown; Animals; Hepatocytes; Humans; Mice; Mitochondria; Muscle Fibers, Skeletal; Phytosterols; Respiration; Sitosterols

Sewage pollution is a principal factor of decreasing water quality, although it has not been considered a real impact in Amazonia that is still considered a pristine environment around the world. Thus, this study aimed to assess the levels of sewage contamination in sediments from three streams crossing Manaus - a Brazilian city of 2,403,796 inhabitants in the heart of the Amazon rain forest. Cholesterol, cholestanol, brassicasterol, ergosterol, stigmasterol, β-sitosterol, campesterol, stigmastanol, coprostanol, and epicoprostanol levels were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS). The fecal indicator, coprostanol, was found in high concentrations (509-12 830 ng g

Topics: Biomarkers; Brazil; Cholestadienols; Cholestanol; Cholestanols; Cholesterol; Chromatography, Liquid; Drug Contamination; Environmental Monitoring; Feces; Geologic Sediments; Phytosterols; Rivers; Sewage; Sitosterols; Sterols; Tandem Mass Spectrometry; Water Pollutants; Water Pollution; Water Quality

There has been an increasing interest during recent years in the role of the gut microbiome on health and disease. Therefore, metabolites in human feces related to microbial activity are attractive surrogate marker to track changes of microbiota induced by diet or disease. Such markers include 5α/β-stanols as microbiome-derived metabolites of sterols. Currently, reliable, robust, and fast methods to quantify fecal sterols and their related metabolites are missing. We developed a liquid chromatography-high-resolution mass spectrometry (LC-MS/HRMS) method for the quantification of sterols and their 5α/β-stanols in human fecal samples. Fecal sterols were extracted and derivatized to N, N-dimethylglycine esters. The method includes cholesterol, coprostanol, cholestanol and sitosterol, 5α/β-sitostanol, campesterol and 5α/β-campestanol. Application of a biphenyl column permits separation of isomeric 5α- and 5β-stanols. Sterols are detected in parallel reaction monitoring (PRM) mode and stanols in full scan mode. HRMS allows differentiation of isobaric β-stanols and the [M + 2] isotope peak of the coeluting sterol. Performance characteristics meet the criteria recommended by Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines. Analysis of fecal samples from healthy volunteers revealed high interindividual variability of sterol and stanol fractions. Interestingly, cholesterol and sitosterol showed similar fractions of mainly 5β-stanols. In contrast, campesterol is substantially converted to 5α-campestanol and might be a poorer substrate for bacterial metabolism. Robust and fast quantification of fecal sterols and their related stanols by LC-MS/HRMS offers great potential to find novel microbiome-related biomarker in large-scale studies.

Topics: Cholesterol; Chromatography, Liquid; Feces; Gastrointestinal Microbiome; Humans; Limit of Detection; Phytosterols; Sitosterols; Sterols; Tandem Mass Spectrometry

Sterols are components present in the fat fraction of infant formulas (IFs). Their characterization is therefore of interest, though there are no official reference methods for their analysis in these matrices.. To validate a gas chromatographic method with flame ionization detection for the determination of animal (cholesterol and desmosterol) and plant sterols (brassicasterol, campesterol, stigmasterol, β-sitosterol and sitostanol) found in IFs. All correlation coefficients obtained for the calibration curves of sterols studied were >0.99. Limits of detection (<1 μg/100 mL) and quantification (<4 μg/100 mL) are suitable for sterols determination in IFs. The within-assay precision ranged from 1.6% to 8.8%, while the between-assay precision was <10% for most of sterols. Accuracy was satisfactory and was calculated by recovery assays (ranging 93-108%). The analytical parameters obtained showed the suitability of the proposed method for the determination of sterols in IFs.

Topics: Calibration; Cholestadienols; Cholesterol; Chromatography, Gas; Desmosterol; Flame Ionization; Infant Formula; Limit of Detection; Phytosterols; Reproducibility of Results; Sitosterols; Stigmasterol

The bioaccessibility (BA) of total and individual plant sterols (PS) of four commercial PS-enriched fermented milk beverages (designated as A to D) was evaluated using in vitro gastrointestinal digestion including the formation of mixed micelles. The fat content of the samples ranged from 1.1 to 2.2% (w/w), and PS enrichment was between 1.5 and 2.9% (w/w). β-Sitosterol, contained in all samples, was higher in samples A and B (around 80% of total PS). The campesterol content was C (22%) > A (7%) > B (5%). Sitostanol was the most abundant in sample D (85%). Stigmasterol was only present in sample C (33%). The greatest BA percentage for total PS corresponded to samples A and B (16-17%), followed by sample D (11%) and sample C (9%). The total BA was not related to the protein, lipid or PS content of the beverages, whereas samples with higher carbohydrates and fiber contents showed lower BA. The BA of the individual PS differed according to the sample considered, and was not related to the PS profile of the sample, thus indicating strong dependency upon the matrix (PS ingredient and other components). Although in vivo studies should be carried out to better assess the functionality of PS in functional foods such as enriched fermented milk beverages, our in vitro study is a useful preliminary contribution to evaluation of the efficacy of these products.

Topics: Biological Availability; Cholesterol; Cultured Milk Products; Dietary Carbohydrates; Dietary Fats; Dietary Fiber; Digestion; Food, Fortified; Functional Food; Gastrointestinal Tract; Micelles; Models, Biological; Phytosterols; Sitosterols; Stigmasterol

A mixture of phytosterols/-stanols, consisting of 75% β-sitosterol, 12% sitostanol, 10% campesterol, 2% campestanol, and 1% others, was esterified with linoleic acid. The resulting mixture of phytosteryl/-stanyl linoleates was subjected to thermal oxidation at 180 °C for 40 min. A silica solid-phase extraction was applied to separate a fraction containing the nonoxidized linoleates and nonpolar degradation products (heptanoates, octanoates) from polar oxidation products (oxo- and hydroxyalkanoates). In total, 15 sitosteryl, sitostanyl, and campesteryl esters, resulting from oxidation of the acyl chain, could be identified by GC-FID/MS. Synthetic routes were described for authentic reference compounds of phytosteryl/-stanyl 7-hydroxyheptanoates, 8-hydroxyoctanoates, 7-oxoheptanoates, 8-oxooctanoates, and 9-oxononanoates, which were characterized by GC-MS and two-dimensional NMR spectroscopy. The study provides data on the formation and identities of previously unreported classes of acyl chain oxidation products upon thermal treatment of phytosteryl/-stanyl fatty acid esters.

Topics: Cholesterol; Esters; Hot Temperature; Linoleic Acids; Molecular Structure; Oxidation-Reduction; Phytosterols; Sitosterols

Ross Lake lies within the City of Flin Flon (Manitoba, Canada), a mining community originally formed by the Hudson Bay Mining and Smelting Company (now Hudbay Minerals Inc.) in 1927. At the time of this investigation, a continuous effluent stream from Hudbay Minerals (approximately 80 years) and a discontinuous and unknown amount of raw and minimally treated municipal sewage (>20 years, likely ending in 1951) was discharged into the north basin of the lake. Maximum concentrations of fecal sterols, such as coprostanol and terrestrial phytosterols, such as: β-sitosterol, campesterol, stigmastanol were measured in vertical sections of sediment cores, collected from Ross Lake, in the 15-16-cm section, which likely corresponds to the 1930s. Concentrations of coprostanol increased from <1 μg g(-1) in older sediments, to 252.3 μg g(-1) organic carbon at the peak. Observed changes in concentrations of sterols, in combination with radiometric dating and changes to sediment physicochemical characteristics, support the conclusion that sediments of a depth of less than 17.5-cm depth were deposited during the post-industrial era from approximately 1930 onwards. Ratios of coprostanol to cholesterol>1, peaking at 3.6 are consistent with anecdotal information that municipal sewage was discharged into Ross Lake during the early years of urbanization, prior to changes in treatment of sewage and discharge practices that began in 1951. Finally, historical concentrations of terrestrial phytosterols followed trends similar to those of coprostanol and cholesterol and may possibly be the result of an increase in the flux of terrestrial organic matter into Ross Lake as the result of regional deforestation due to logging and fire.

Topics: Cholesterol; Environmental Monitoring; Feces; Geologic Sediments; History, 20th Century; History, 21st Century; Lakes; Manitoba; Phytosterols; Sewage; Sitosterols; Waste Disposal, Fluid; Water Pollutants; Water Purification

This study has determined oil, fatty acid (FA) and phytosterols content during the ripening of the Tunisian Onopordum acanthium L. seeds. In total, nine FAs and six phytosterols were identified. The main FAs were linoleic acid (0.18-8.06 mg/g of seed) followed by oleic acid (0.051-2.45 mg/g of seed), palmitic acid and stearic acid. Pentadecanoic acid was detected, for the first time, in unripe fruits and the two last stages of development were characterised by a relative abundance of erucic acid. Overall, β-sitosterol (34.5-77.79% of total sterols) was the major 4-desmethylsterols during maturation. The first episodes of growth were characterised by the best amounts of stigmasterol and campesterol, while stigmastanol and Δ7 sitosterol had quoted the semi-ripe and fully ripe fruits; however, cholesterol was absent. These findings are useful in understanding a potential new source of important natural compounds (Phytosterols and USFA) found in this fruit and when harvest should be undertaken to optimise desired FA and phytosterols content.

Topics: Cholesterol; Fatty Acids; Fruit; Linoleic Acid; Oleic Acid; Onopordum; Phytosterols; Plant Oils; Seeds; Sitosterols; Stigmasterol; Tunisia

The present study was to evaluate the cholesterol-lowering effect of two novel plant stanol derivatives and its potential molecular mechanism in hyper-cholesterol mice induced by a high-cholesterol diet. Results showed that oral administration of plant stanyl hemisuccinate (2×, 5×) and plant stanyl sorbitol succinate (2×, 5×) effectively attenuated the serum total cholesterol and low density lipoprotein cholesterol levels, while had no effect on the serum triacylglycerol and high density lipoprotein cholesterol. And plant stanol derivatives decreased liver cholesterol concentration and increased faecal cholesterol output. Meanwhile, both plant stanyl hemisuccinate and plant stanyl sorbitol succinate could remarkably promote liver X receptor alpha (LXRα) expression, and increased cholesterol 7α-hydroxylase (CYP7A1) expression and faecal total bile acid output to varying degrees. These results suggested two novel plant stanol derivatives possessed hypocholesterolemic effect, and the cholesterol-lowering action of plant stanol derivatives may be through activating the potential LXRα-CYP7A1-bile acid excretion pathway.

Topics: Animals; Cholesterol; Cholesterol 7-alpha-Hydroxylase; Gene Expression; Hydroxymethylglutaryl CoA Reductases; Liver; Liver X Receptors; Male; Mice; Orphan Nuclear Receptors; Phytosterols; Plant Extracts; Sitosterols; Sterol Regulatory Element Binding Protein 1

The rate of APC mutations in the intestine increases in middle-age. At the same period of life, plant sterol and stanol enriched functional foods are introduced to diet to lower blood cholesterol. This study examined the effect of plant stanol enriched diet on intestinal adenoma formation in the Apc(Min) mouse. Apc(Min) mice were fed 0.8% plant stanol diet or control diet for nine weeks. Cholesterol, plant sterols and plant stanols were analyzed from the caecum content and the intestinal mucosa. Levels of β-catenin, cyclin D1, epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase 1/2 (ERK1/2) were measured from the intestinal mucosa by Western blotting. Gene expression was determined from the intestinal mucosa using Affymetrix and the data were analyzed for enriched categories and pathways. Plant stanols induced adenoma formation in the small intestine, however, the adenoma size was not affected. We saw increased levels of nuclear β-catenin, phosphorylated β-catenin (Ser675 and Ser552), nuclear cyclin D1, total and phosphorylated EGFR and phosphorylated ERK1/2 in the intestinal mucosa after plant stanol feeding. The Affymetrix data demonstrate that several enzymes of cholesterol synthesis pathway were up-regulated, although the cholesterol level in the intestinal mucosa was not altered. We show that plant stanols induce adenoma formation by activating Wnt and EGFR signaling. EGFR signaling seems to have promoted β-catenin phosphorylation and its translocation into the nucleus, where the expression of cyclin D1 was increased. Up-regulated cholesterol synthesis may partly explain the increased EGFR signaling in the plant stanol-fed mice.

Topics: Adenoma; Animals; beta Catenin; Cecum; Cholesterol; Cyclin D1; ErbB Receptors; Female; Gene Expression Regulation; Genes, APC; Intestinal Mucosa; Intestinal Neoplasms; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Mitogen-Activated Protein Kinase 3; Phytosterols; Proto-Oncogene Proteins c-akt; Serine; Sitosterols; Sterol Regulatory Element Binding Protein 2; Up-Regulation; Wnt Signaling Pathway

Improving the microbiological quality of coastal and river waters relies on the development of reliable markers that are capable of determining sources of fecal pollution. Recently, a principal component analysis (PCA) method based on six stanol compounds (i.e. 5β-cholestan-3β-ol (coprostanol), 5β-cholestan-3α-ol (epicoprostanol), 24-methyl-5α-cholestan-3β-ol (campestanol), 24-ethyl-5α-cholestan-3β-ol (sitostanol), 24-ethyl-5β-cholestan-3β-ol (24-ethylcoprostanol) and 24-ethyl-5β-cholestan-3α-ol (24-ethylepicoprostanol)) was shown to be suitable for distinguishing between porcine and bovine feces. In this study, we tested if this PCA method, using the above six stanols, could be used as a tool in "Microbial Source Tracking (MST)" methods in water from areas of intensive agriculture where diffuse fecal contamination is often marked by the co-existence of human and animal sources. In particular, well-defined and stable clusters were found in PCA score plots clustering samples of "pure" human, bovine and porcine feces along with runoff and diluted waters in which the source of contamination is known. A good consistency was also observed between the source assignments made by the 6-stanol-based PCA method and the microbial markers for river waters contaminated by fecal matter of unknown origin. More generally, the tests conducted in this study argue for the addition of the PCA method based on six stanols in the MST toolbox to help identify fecal contamination sources. The data presented in this study show that this addition would improve the determination of fecal contamination sources when the contamination levels are low to moderate.

Topics: Animals; Cattle; Cholestanes; Cholestanol; Cholestanols; Feces; Fresh Water; Humans; Phytosterols; Principal Component Analysis; Rivers; Seawater; Sitosterols; Swine; Water Microbiology; Water Pollutants, Chemical

An in vitro lipolysis model was utilized to study the effect of stigmastanol (lipophilic phytosterol) and disodium ascorbyl phytostanol phosphate (DAPP) (modified hydrophilic phytostanol) on intestinal processing of cholesterol to gain further understanding of their cholesterol lowering mechanism. Lipolysis results showed that stigmastanol, if given in powder alone, had no effect on cholesterol processing probably due to its poor solubility. Stigmastanol suspension formulation re-distributed cholesterol from aqueous phase to oil and sediment phases. The water soluble DAPP has changed cholesterol distribution even more significantly by transferring cholesterol from aqueous phase to sediment phase. Moreover, the results provided evidence that DAPP inhibited triglyceride digestion in vitro. Considering DAPP as a surfactant with the same lipophilic sterol ring as bile salt, its ability to inhibit triglyceride lipolysis may be due to its competition with bile salt for the substrate surface, thereby hindering the lipolysis of triglyceride and inhibiting cholesterol solubilization with the lipolysis products. It can be speculated that the cholesterol lowering mechanism of DAPP during intestinal digestion is related to its ability to act as a surfactant closely resembling bile salt.

Topics: Cholesterol; Intestinal Mucosa; Lipolysis; Models, Biological; Phytosterols; Sitosterols

Triterpenoid compounds found in free and ester forms in extracts of entire fruits and leaves and in fruit and leaf cuticular waxes of bilberry (Vaccinium myrtillus L.) collected in Finland and Poland were identified and quantitated by gas chromatography-mass spectrometry coupled to a flame ionization detector (GC-MS/FID). The main bilberry triterpenoid profile consisted of α- and β-amyrin, α- and β-amyrenone, campesterol, cholesterol, citrostadienol (in berries), cycloartanol, erythrodiol, lupeol, 24-methylenecycloartanol, sitosterol, sitostanol, stigmasterol, stigmasta-3,5-dien-7-one, uvaol, oleanolic and ursolic aldehydes, and oleanolic, ursolic, 2α-hydroxyoleanolic, and 2α-hydroxyursolic acids. Friedelin and D:A-friedooleanan-3β-ol were found only in Finnish plants, whereas D:C-friedours-7-en-3β-ol and taraxasterol were found only in Polish plants. To our knowledge, this is the first thorough description of triterpenoid compounds in this species. The presented results revealed that the triterpenoid profile of bilberry varied considerably between different organs of the plant, regardless of the plant origin, as well as between plant samples obtained from the two geographical locations.

Topics: Anthocyanins; Cholesterol; Finland; Fruit; Gas Chromatography-Mass Spectrometry; Molecular Structure; Oleanolic Acid; Phytosterols; Plant Extracts; Plant Leaves; Poland; Sitosterols; Triterpenes; Vaccinium myrtillus

Plant sterols may induce a Th1 shift in humans. However, whether plant stanols have similar effects as well as the underlying mechanism are unknown. We have now shown that (like sitosterol) sitostanol, both 4-desmethylsterols, induces a Th1 shift when added in vitro at physiological concentrations to human PBMCs. This conclusion was based on a higher IFNgamma production, with no change in the production of IL-4 and IL-10. alpha-Amyrin, a 4.4-dimethylsterol, had comparable effects. Because 4.4-dimethylsterols cannot activate transcription factor LXR, this finding indicates that LXR activation was not involved. Sitosterol and sitostanol did not alter the production of IL-12 and IL-18 in PBMCs as well as in monocyte-derived U937 cells, suggesting that plant sterols directly affect T-helper cells, without activating APCs. However, in PBMCs treated with a TLR2 blocker (T2.5), IFNgamma production was completely inhibited, whereas blocking TLR4 with HTA125 had no such effect. To confirm these findings, PBMCs from TLR2(-/-) mice were cultured in the presence of sitosterol and sitostanol. In these cells, no Th1 shift was observed. Our results, therefore, indicate that TLR2 activation is essential to induce a Th1 shift in human PBMCs by plant stanols and plant sterols.

Topics: Animals; Humans; Interferon-gamma; Interleukin-12; Interleukin-18; Leukocytes, Mononuclear; Liver X Receptors; Mice; Mice, Inbred C57BL; Mice, Transgenic; Orphan Nuclear Receptors; Phytosterols; Plants; Sitosterols; Th1 Cells; Toll-Like Receptor 2; U937 Cells

The unsaponifiable fraction of six Tunisian monovarietal virgin olive oils from the region of Medenine was evaluated within a single chromatographic run by using HPLC-APCI-tandem MS. Separation of the compounds under study was achieved by the RP-LC method, giving a reasonable analysis time and good resolution. Detection was done by an ion trap (working alternatively in MS and MS/MS modes), the fact which made our method suitable to unequivocally identify a high number of compounds belonging to different families of the unsaponifiable fraction of oil and to carry out their reliable and sensitive quantification. A great amount of qualitative information was generated in every analysis, although we focused on the quantification of sterols, tocopherols, and triterpenic dialcohols since their standards were commercially available. The limits of detections achieved were within the range of 1.21 and 10.31 microg/kg for sitostanol and beta-sitosterol, respectively. Significant differences were observed in the composition of the studied olive cultivars. Jemri Ben Guerdane oil was the richest one in terms of all of the sterols under study. alpha-Tocopherol was the main vitamin E isomer in all samples, ranging from 70.14 to 130.72 mg/kg. Principal component analysis (PCA) and cluster analysis were applied to the whole data set in order to explore the distribution of the olive cultivars according to their oil composition.

Topics: Alcohols; Chromatography, High Pressure Liquid; Olive Oil; Phytosterols; Plant Oils; Saponins; Sitosterols; Species Specificity; Tandem Mass Spectrometry; Tocopherols; Triterpenes; Tunisia

Much interest has focused on the cholesterol-lowering effects of phytosterols (plant sterols) but limited data suggests they may also possess anti-carcinogenic activity. Conjugated linoleic acids (CLA), sourced from meat and dairy products of ruminant animals, has also received considerable attention as a potential anti-cancer agent. Therefore, the aims of this project were to (i) examine the effects of phytosterols and CLA on the viability and growth of human intestinal Caco-2 cells and (ii) determine their potential genoprotective (comet assay), COX-2 modulatory (ELISA) and apoptotic (Hoechst staining) activities. Caco-2 cells were supplemented with the phytosterols campesterol, beta-sitosterol, or beta-sitostanol, or a CLA mixture, or individual CLA isomers (c10t12-CLA, t9t11-CLA) for 48 h. The three phytosterols, at the highest levels tested, were found to reduce both the viability and growth of Caco-2 cells while CLA exhibited isomer-specific effects. None of the phytosterols protected against DNA damage. At a concentration of 25 microM, both c10t12-CLA and t9t11-CLA enhanced (P<0.05) oxidant-induced, but not mutagen-induced, DNA damage. Neither the phytosterols nor CLA induced apoptosis or modulated COX-2 production. In conclusion, campesterol, beta-sitosterol, beta-sitostanol, c10t12-CLA, and t9t11-CLA were not toxic to Caco-2 cells, at the lower levels tested, and did not exhibit potential anti-carcinogenic activity.

Topics: Caco-2 Cells; Cell Membrane; Cell Survival; Cholesterol; Comet Assay; Cyclooxygenase 2; DNA Damage; Enzyme-Linked Immunosorbent Assay; Humans; Hydrogen Peroxide; L-Lactate Dehydrogenase; Linoleic Acid; Methylnitronitrosoguanidine; Mutagens; Phytosterols; Protective Agents; Sitosterols

To analyze the phytosterol content in food plant materials and Chinese traditional herbal medicines commonly used in China.. 18 kinds of food plant materials and 32 kinds of Chinese traditional herbal medicines, which were commonly used in functional food, were chosen as samples. The contents of beta-sitosterol, campesterol, stigmasterol, beta-sitostanol were analyzed by GC methods and the percent of each ingredient were calculated.. The contents of phytosterols in 18 kinds of food plant materials were from 14.8 mg/100 g to 208.3 mg/100 g, while the content of phytosterols in 32 Chinese traditional herbal medicines were from 9.4 mg/100 g to 280.3 mg/100 g. In most samples, beta-sitosterol is the largest part of total phytosterol.. Phytosterols were existed in 50 kinds of food plant materials and Chinese traditional herbal medicines commonly used in functional food, maybe phytosterol is an important functional ingredient in some plant materials.

Topics: Cholesterol; Chromatography, Gas; Drugs, Chinese Herbal; Phytosterols; Plants, Medicinal; Sitosterols; Stigmasterol; Vegetables

Rapeseed oil (RSO) is a novel source of plant sterols, containing the unique brassicasterol in concentrations higher than allowed for plant sterol blends in food products in the European Union. Effects of RSO sterols and stanols on aortic atherosclerosis were studied in cholesterol-fed heterozygous Watanabe heritable hyperlipidaemic (Hh-WHHL) rabbits. Four groups (n 18 per group) received a cholesterol-added (2 g/kg) standard chow or this diet with added RSO stanol esters (17 g/kg), RSO stanol esters (34 g/kg) or RSO sterol esters (34 g/kg) for 18 weeks. Feeding RSO stanol esters increased plasma campestanol (P < 0.001) and sitostanol (P < 0.001) and aortic campestanol (P < 0.05) compared with controls. Feeding RSO sterol esters increased concentrations of plasma campesterol (P < 0.001), sitosterol (P < 0.001) and brassicasterol (P < 0.001) and aortic campesterol (P < 0.01). Significantly lower plasma cholesterol (P < 0.001) was recorded in the treated groups after 3 weeks and throughout the study. LDL-cholesterol was reduced 50 % in the high-dose RSO sterol ester (P < 0.01) and high-dose RSO stanol ester (P < 0.001) groups compared with controls. Atherosclerotic lesions were found in three rabbits in each of the RSO stanol ester groups and in one in the RSO sterol ester group. Aortic cholesterol was decreased in the treated groups (P < 0.001) in response to lowering of plasma cholesterol induced by RSO sterol and stanol esters. In conclusion, RSO stanol and sterol esters with a high concentration of brassicasterol were well tolerated. They were hypocholesterolaemic and inhibited experimental atherosclerosis in cholesterol-fed Hh-WHHL rabbits. A significant uptake of plant sterols into the blood and incorporation of campesterol and campestanol into aortic tissue was recorded.

Topics: Animals; Aorta; Atherosclerosis; Cholestadienols; Cholesterol; Cholesterol, Dietary; Fatty Acids, Monounsaturated; Female; Heterozygote; Hyperlipidemias; Lipids; Male; Phytosterols; Plant Oils; Rabbits; Rapeseed Oil; Sitosterols

We elucidated the molecular mechanism(s) underlying sterol trafficking by investigating alterations in gene expression in response to increased retention of dietary phytosterols and phytostanols in stroke-prone spontaneously hypertensive (SHRSP) and normotensive Wistar Kyoto (WKY) inbred rats.. SHRSP and WKY inbred rats were fed a control diet or a diet supplemented with phytosterols or phytostanols (2 g/kg diet).. Intake of phytosterols and phytostanols increased their incorporation in plasma, red blood cells, liver, aorta and kidney, but decreased cholesterol levels in liver and aorta in both rat strains. Phytosterol intake up-regulated mRNA expression of intestinal Npc1l1 and Abcg8, and hepatic Abcg5, Abca1, Cyp27a1 and Hmgcr. Phytostanol intake up-regulated Npc1l1 and Srebp2, but down-regulated Abcg5 mRNA expression in small intestine. Phytostanols also up-regulated Abca1 expression in SHRSP rats, but down-regulated Abca1 expression in WKY inbred rats. Compared to phytosterols, dietary phytostanols reduced phytosterol levels in plasma, red blood cells, and kidney, as well as altered mRNA levels of hepatic Abca1,Cyp27a1, and Hmgcr and intestinal Abcg5/8, Hmgcr and Srebp2.. Altered expression of multiple sterol-regulatory genes may contribute to the incorporation and cholesterol-lowering actions of phytosterols and phytostanols. Phytosterols and phytostanols may act through different mechanism(s) on cholesterol and phytosterol/phytostanol trafficking.

Topics: Animals; Anticholesteremic Agents; Cholestadienols; Cholesterol; Gene Expression Regulation; Hypolipidemic Agents; Jejunum; Liver; Male; Organ Specificity; Phytosterols; Rats; Rats, Inbred SHR; Rats, Inbred WKY; RNA, Messenger; Sitosterols; Sterols

To investigate the dietary phytosterol intake of elderly women in three different cities of China, and to compare the main dietary sources, so that to discuss the relationship of dietary phytosterol intake and serum lipids.. Based on the dietary pattern, women more than 50 years old from Beijing, Hefei and Urumchi were chosen as testers, 80 - 100 people for each city respectively. The dietary survey was done by continues 24 hours review of two days, the plant food were collected and the phytosterol content (include beta-sitosterol, campesterol, stigmasterol, sitostanol) were analyzed by GC methods, the total phytosterols content were calculated. The dietary phytosterol intake were calculated and serum lipids were also analyzed in all the testers.. Testers from Beijing, Hefei and Urumchi were 100, 101 and 84 respectively. The average dietary phytosterol intake of people in Beijing and Hefei were 340.3 mg/d and 313.5 mg/d, the main sources were plant oil and cereals, while the average dietary phytosterol intake of people in Urumchi were 550.4 mg/d, higher than the other two cities (t values were 9.369, 10.420, respectively, both P values < 0.01), the main source in Urumchi was cereal (provide 53.1% of the total phytosterol intake). The laboratory results showed, testers in Urumchi had significantly lower serum TC content ((4.04 +/- 0.78) mmol/L) than that in Beijing ((4.89 +/- 0.91) mmol/L) and Hefei ((4.71 +/- 0.83) mmol/L) (t value were 6.766 and 5.401 respectively, both P values < 0.01); serum TG content in Urumchi((1.01 +/- 0.48) mmol/L) was also lower than that in Beijing ((1.31 +/- 0.53) mmol/L) and Hefei ((1.66 +/- 0.75) mmol/L) (t values were 3.343 and 7.293 respectively, both P values < 0.01); the serum glucose is also lower in testers in Urumchi ((5.02 +/- 2.18) mmol/L) compared with testers in Beijing ((5.69 +/- 1.53) mmol/L, t = 2.561, P < 0.05) and Hefei ((5.78 +/- 1.53) mmol/L, t = 2.934, P < 0.01).. Different dietary pattern result in significantly different dietary phytosterol intake in elder women in three cities, higher, phytosterol intake seemed to contribute to lower serum lipids.

Topics: Aged; Aged, 80 and over; China; Cholesterol; Cholesterol, Dietary; Female; Humans; Lipids; Middle Aged; Phytosterols; Sitosterols; Urban Population

We investigated the difference between the molecular structures of plant sterols and stanols that affect the solubilization of cholesterol in bile salt micelles (in vitro study). First, the aqueous solubility of beta-sitosterol, beta-sitostanol, and campesterol was determined by considering the specific radioactivity by using a fairly small quantity of each radiolabeled compound. The order of their aqueous solubilities was as follows: cholesterol > campesterol > beta-sitostanol > beta-sitosterol. The maximum solubility of cholesterol and the above mentioned sterol/stanol in sodium taurodeoxycholate and sodium taurocholate solutions (single solubilizate system) was measured. Moreover, the preferential solubilization of cholesterol in bile salt solutions was systematically studied by using different types of plant sterols/stanols. The solubilization results showed that the cholesterol-lowering effect was similar for sterols and stanol. Thermodynamic analysis was applied to these experimental results. The Gibbs energy change (Delta G degrees ) for the solubilization of plant sterols/stanols showed a negative value larger than that for cholesterol.

Topics: Cholesterol; Micelles; Phytosterols; Sitosterols; Solubility; Taurocholic Acid; Taurodeoxycholic Acid; Thermodynamics

Functional foods with supplementation of plant sterols are already used by millions of people. However, at the same time it is current scientific thinking that elevation of plant sterols in the circulation causes coronary heart disease. Therefore, this study aimed to define the risk for coronary heart disease associated with moderately high plant sterol plasma levels in a cohort of elderly. In this study, we evaluated the association between plant sterols and coronary heart disease in a cohort of 1242 subjects older than 65 years, participating at the Longitudinal Aging Study Amsterdam (LASA). Concentrations of sitosterol, campesterol, brassicasterol and stigmasterol were assessed using highly sensitive and specific gas chromatography-mass spectrometry-selected ion-monitoring. Plant sterol concentrations (and their ratios to cholesterol) were slightly, however, significantly lower in patients with coronary heart disease. Moreover, high plasma concentrations of a marker plant sterol, sitosterol, were associated with a markedly reduced risk for coronary heart disease (OR 0.78, CI 0.62-0.98, p<0.05). In contrast neither plant stanols (sitostanol or campestanol) nor the cholesterol synthesis markers (lathosterol, lanosterol and desmosterol) nor their ratios to cholesterol were significantly different in the study groups. These data suggest that plant sterols could have neutral or even protective effects on development of coronary heart disease, which have to be confirmed in interventional trials.

Topics: Aged; Aged, 80 and over; Cholesterol; Coronary Disease; Cross-Sectional Studies; Female; Humans; Logistic Models; Male; Peripheral Vascular Diseases; Phytosterols; Risk Factors; Sitosterols

The aim of this study was to investigate the cholesterol-lowering mechanisms of corn fiber oil (CFO), ferulate phytostanyl esters (FPEs) and parent compounds of FPE, including sitostanol and ferulic acid, in hamsters.. Seventy male Golden Syrian hamsters were randomly assigned to six experimental diets for 4 weeks: (1) cornstarch-casein-sucrose-based control diet (control); and (2) control diet plus 0.1% (wt/wt) cholesterol (cholesterol-control). The remaining four groups were given cholesterol-control diet with: (3) 10% (wt/wt) CFO; (4) 0.5% (wt/wt) sitostanol; (5) 0.23% (wt/wt) ferulic acid; and (6) 0.73% (wt/wt) FPE. At the end of dietary intervention, total plasma cholesterol, high-density lipoprotein cholesterol and triglyceride concentrations were determined. Parameters of cholesterol kinetics, including cholesterol absorption and synthesis, as well as mRNA expression of sterol transporters such as Niemann-Pick C1 like 1 (NPC1L1), ATP-binding cassette G5 (ABCG5) and ABCG8, were assessed.. Supplementation with CFO decreased (P<.0001) plasma total cholesterol levels by 29% as compared with the cholesterol-control group, while FPE and sitostanol reduced (P<.02) cholesterolemia by 15% and 14%, respectively. CFO and sitostanol decreased (P<.05) cholesterol absorption by 24% compared to the cholesterol-control group. Dietary intervention did not alter the intestinal gene expression of ABCG5, ABCG8 and NPC1L1.. The present results show that the CFO-induced and sitostanol-induced decrease in cholesterol absorption is independent of intestinal enterocyte sterol transporters such as ABCG5, ABCG8 and NPC1L1 in hamsters.

Topics: Animals; Anticholesteremic Agents; ATP-Binding Cassette Transporters; Cholesterol; Corn Oil; Coumaric Acids; Cricetinae; Intestinal Absorption; Intestinal Mucosa; Male; Membrane Transport Proteins; Phytosterols; Sitosterols; Sterols

The objective of the present study was to determine the beneficial effects and the safety of oral administration of the combination of berberine (BBR) and plant stanols (PS) on plasma lipid profiles in male Sprague-Dawley rats. Four groups of animals were fed a cornstarch-casein-sucrose-based high-cholesterol (2%, w:w) and high-fat (27.5%) diet. Three treatment groups were supplemented with either BBR (100mgkg(-1)bodyweightd(-1)), PS (1% in diet, w:w), or the combination of both (BBRPS). After 6 wk, animals were sacrificed and followed immediately with the collection of blood and organ samples. Lipid analysis revealed that PS lowered plasma total cholesterol (T-C) by 18% (p=0.067) and non-HDL-cholesterol (non-HDL-C) by 29% (p=0.013) as compared with the control, while BBR had no effect on both T-C and non-HDL-C. The combination treatment of BBRPS reduced plasma T-C by 41% (p=0.0002) and non-HDL-C by 59% (p<0.0001) compared to the control group. BBR reduced plasma TG levels by 31% at a marginal significance relative to the control (p=0.054), whereas PS had no effect. BBRPS showed an additive effect of BBR and PS on plasma TAG. PS and BBRPS both decreased liver cholesterol (p=0.0027 and 0.0002, respectively). BBR and PS, either alone or in combination, did not show any toxic effects as assessed by plasma concentration of hepatic biochemical parameters. These results demonstrate that BBR and PS, when combined, synergistically lower plasma cholesterol levels and significantly reduce liver cholesterol, without the observation of any toxic effects.

Topics: Administration, Oral; Animals; Anticholesteremic Agents; Berberine; Cholesterol; Diet, Atherogenic; Drug Therapy, Combination; Lipoproteins; Male; Phytosterols; Rats; Rats, Sprague-Dawley; Sitosterols; Triglycerides

The plant sterols campesterol, beta-sitosterol and beta-sitostanol were investigated for potential immunomodulatory effects in Jurkat T cells. Treatments involved supplementing cells with or without concanavalin A (ConA) or phorbol-12-myristate-13-acetate plus ionomycin (PMA+IoM) in the presence or absence of increasing concentrations (10-100 microM) of each plant sterol for 24 h. None of the plant sterols significantly affected mitogen-stimulated IL-4, IL-10 or IFN-gamma production. However, campesterol, beta-sitosterol and beta-sitostanol significantly suppressed mitogen-induced IL-2 production in a dose-dependent manner. Both bisindolylmaleimide-I (BIM-I), a specific protein kinase C (PKC) inhibitor, and the immunosuppressant drug known as Tacrolimus (FK506), an IL-2 inhibitor, prevented mitogen-stimulated IL-2 production in Jurkat cells. Treatment with PMA+IoM alone significantly increased PKC activity and the presence of BIM-I prevented PKC activation by PMA+IoM. Following 24 h treatments, the plant sterols did not affect PMA+IoM-enhanced PKC activity, cellular calcium content or calcineurin activity. Intracellular cyclic 3',5'-adenosine monophosphate (cAMP) levels were significantly reduced by PMA+IoM. The presence of FK506 prevented a PMA+IoM-induced reduction of intracellular cAMP. Likewise the plant sterols behaved in a similar manner as FK506. Our findings suggest that the suppression of IL-2 by the plant sterols was not mediated via PKC inhibition and that their effects occurred possibly via cAMP modulation and/or a calcium/calcineurin-independent pathway.

Topics: Cell Division; Cell Survival; Cholesterol; Concanavalin A; Cytokines; Enzyme Inhibitors; Humans; Immunologic Factors; Immunosuppressive Agents; Indoles; Interleukin-2; Jurkat Cells; Lymphocyte Activation; Maleimides; Phytosterols; Protein Kinase C; Sitosterols; T-Lymphocytes; Tacrolimus; Tetradecanoylphorbol Acetate

To analyze the phytosterol content in plant food commonly consumed in China, and to estimate the intake of phytosterols in Chinese people.. More than 160 types of plant food in 7 kinds were chosen as samples. The contents of beta-sitosterol, campesterol, stigmasterol, beta-sitostanol, campestanol were analyzed by GC methods and the total phytosterols were calculated. The intake of phytosteols in Chinese people was estimated using the data of "Survey on the Status of Nutrition and Health of the Chinese People" in 2002.. The contents of phytosterols in edible oils, nuts, and soybeans were higher than those in other plant food. In cereals, phytosterol contents of wheat flour were much higher than those of rice, the refinements of cereals may decrease the phytosterol contents. The phytosterol contents in vegetables and fruits were lower. The total intake of phytosterols in Chinese people was estimated to be 322.41mg/day, in which 40% may be of edible oil origin and 40% may be of cereal origin.. The results indicated that in the current dietary pattern, increase the intake of wheat, soybean, vegetable and fruit would enhance the phytosterol intake in Chinese.

Topics: China; Fabaceae; Food Analysis; Humans; Oryza; Phytosterols; Sitosterols; Triticum; Vegetables

Plant stanols and sterols of the 4-desmethyl family (e.g., sitostanol and sitosterol) effectively decrease LDL cholesterol concentrations, whereas 4,4-dimethylsterols (alpha-amyrin and lupeol) do not. Serum carotenoid concentrations, however, are decreased by both plant sterol families. The exact mechanisms underlying these effects are not known, although effects on micellar composition have been suggested. With a liver X receptor (LXR) coactivator peptide recruitment assay, we showed that plant sterols and stanols from the 4-desmethylsterol family activated both LXRalpha and LXRbeta, whereas 4,4-dimethyl plant sterols did not. In fully differentiated Caco-2 cells, the functionality of this effect was shown by the increased expression of ABCA1, one of the known LXR target genes expressed by Caco-2 cells in measurable amounts. The LXR-activating potential of the various plant sterols/stanols correlated positively with ABCA1 mRNA expression. Reductions in serum hydrocarbon carotenoids could be explained by the effects of the 4-desmethyl family and 4,4-dimethylsterols on micellar carotenoid incorporation. Our findings indicate that the decreased intestinal absorption of cholesterol and carotenoids by plant sterols and stanols is caused by two distinct mechanisms.

Topics: Antioxidants; ATP Binding Cassette Transporter 1; ATP-Binding Cassette Transporters; Caco-2 Cells; Carotenoids; Cholesterol; Cholesterol, LDL; DNA-Binding Proteins; Humans; Hydrocarbons; Intestinal Absorption; Intestines; Liver X Receptors; Micelles; Models, Chemical; Oleanolic Acid; Orphan Nuclear Receptors; Pentacyclic Triterpenes; Peptides; Phytosterols; Plant Extracts; Receptors, Cytoplasmic and Nuclear; Receptors, Steroid; RNA, Messenger; Sitosterols; Sterol Regulatory Element Binding Protein 2; Triterpenes

The formation of mixed water-insoluble poorly absorbable crystals between cholesterol (CH) and phytosterols (PS) or phytostanols (PSS) in the intestinal lumen has been considered for a long time as a plausible mechanism of the PS/PSS-induced reduction of serum CH concentration. In this report, we demonstrated with the use of the powder X-ray diffraction (XRD) and the differential scanning calorimetry (DSC) techniques that mixed CH:beta-sitosterol (SI) crystals can be formed by recrystallization of corresponding mixtures from melts and also from mixed CH:SI solutions in triglyceride oil. Formation of mixed CH:SI crystals takes place in a wide interval of CH:SI ratios, from approximately 10 up to approximately 75 wt.% of SI in the mixture. Formation of mixed CH:sitostanol (SS) crystals from melts and solutions in triglyceride oil was also detected, but in a more narrow interval of CH:SS ratios. However, during the lipolysis of model dietary emulsions under in vitro conditions, the formation of crystalline material was not detected due to the relatively high solubility of free sterols/stanols in products of fat hydrolysis. We found that the solubility of free CH, SI, and SS raises upon the increase in the solvent polarity, i.e. free fatty acid > diglycerideoil > triglyceride oil. Therefore, we believe that the cocrystallization mechanism of phytosterol-induced serum CH lowering has relatively low importance, unless the diet is specially designed to include relatively little amounts of dietary fats. The presented experimental evidence demonstrates that it is unlikely that the formation of poorly absorbable mixed crystals largely affects the intestinal absorption of CH and, therefore, that this is a prime mechanism by which PS and PSS effect CH absorption.

Topics: Absorption; Calorimetry, Differential Scanning; Cholesterol; Crystallization; Lipase; Phytosterols; Sitosterols; Solubility; Triglycerides; X-Ray Diffraction

Factors contributing to the oxidative stability of phytosterols were studied. Unsaturated stigmasterol and saturated sitostanol were used as model compounds and were heated at different temperatures in different lipid matrices for various periods of time. Accumulations of the major secondary oxidation products were used as a marker of the stability of heated compounds, and the products were analyzed by gas chromatography-mass spectrometry. The results showed that both temperature and heating time, as well as sterol structure and lipid matrix composition, affected phytosterol oxidation. In particular, the interactions between different lipid matrices and temperatures had drastic effects on the total contents of the phytosterol oxides formed and also on the reaction pathways of oxidation. During heating at high temperatures for prolonged periods, >20% of stigmasterol was oxidized. At moderate temperatures the oxidation of stigmasterol was rather slow. Sitostanol oxide contents were low under all heating conditions studied.

Topics: Drug Stability; Food Handling; Gas Chromatography-Mass Spectrometry; Hot Temperature; Lipids; Oxidation-Reduction; Phytosterols; Sitosterols; Stigmasterol; Time Factors

The recent identification of the aberrant transport proteins ABCG5 and ABCG8 resulting in sitosterolemia suggests that intestinal uptake of cholesterol is an unselective process, and that discrimination between cholesterol and plant sterols takes place at the level of sterol efflux from the enterocyte. Although plant sterols are structurally very similar to cholesterol, differing only in their side chain length, they are absorbed from the intestine to a markedly lower extent. In order to further evaluate the process of discrimination, three different sterols (cholesterol, campesterol, sitosterol) and their corresponding 5 alpha-stanols (cholestanol, campestanol, sitostanol) were compared concerning their concentration in the proximal small intestine, in serum, and in bile after a single oral dose of deuterated compounds. The data obtained support the hypothesis that i) the uptake of sterols and stanols is an extremely rapid process, ii) discrimination probably takes place on the level of reverse transport back into the gut lumen, iii) plant stanols are taken up, but not absorbed to a measurable extent, and iv) the process of discrimination probably also exists at the level of biliary excretion. The range of structural alterations that decrease intestinal absorption and increase biliary excretion is: 1) campesterol, 2) cholestanol-sitosterol, and 3) campestanol-sitostanol.

Topics: Animals; Bile; Cholestanols; Cholesterol; Cholesterol, Dietary; Intestinal Absorption; Intestine, Small; Male; Mice; Phytosterols; Plant Extracts; Sitosterols

Plant sterol and stanol esters were separated on a Luna hexyl-phenyl column using a gradient of acetonitrile (90-100%) in water. The eluted compounds were detected by atmospheric pressure chemical ionization (APCI)-mass spectroscopy (MS) in the positive mode. Sterol and stanol esters produced [M + H - HOOCR](+) ions. Application of the hyphenated technique-LC-MS-allowed differentiation between a number of esters of sitosterol, campesterol, stigmasterol, and (tentatively) avenasterol, as well as sitostanol and campestanol esters. With cholesteryl decanoate used as the internal standard, the method showed good linearity, precision, and reproducibility. The method required minimal sample pretreatment and can be applied to samples with high water content (juices) as well as samples with high oil content (margarine spreads). The method could be useful for the analysis of sterol and stanol esters in fortified food products.

Topics: Anticholesteremic Agents; Beverages; Cholesterol; Chromatography, High Pressure Liquid; Citrus; Esters; Fruit; Margarine; Mass Spectrometry; Phytosterols; Sensitivity and Specificity; Sitosterols; Stigmasterol

Campesterol is present in all the phytosterol-containing dietary hypocholesterolemic agents in current use. Campesterol is absorbed more efficiently than sitosterol, and the question of its possible atherogenicity has been raised. To test this possibility, rabbits were fed either a semipurified, cholesterol-free diet that has been shown to be atherogenic for this species or the same diet augmented with 0.5 g of phytosterol-rich diet preparations (spreads) containing either sitosterol or sitostanol. The diets contained 295 mg phytosterol per 100 g. After 60 d, serum cholesterol levels in the two phytosterol groups were 78 +/- 4 mg/dL (sitosterol) and 76 +/- 4 mg/dL (sitostanol), respectively. The serum cholesterol level of rabbits fed the control diet was 105 +/- 8 mg/dL. Serum campesterol (microg/mL) levels were higher than sitosterol or sitostanol levels in all groups. Aortic phytosterols were present in nanogram quantities compared to cholesterol, which was present in microgram quantities. The ratio of campesterol/sitosterol/sitostanol in the aortas was: control, 1.00:0.43:0.02; sitosterol, 1:00:0.32:0.01; sitostanol, 1:00:0.34:0.11. Aortic campesterol was present at 4% the concentration of aortic cholesterol, sitosterol at 1.4%, and sitostanol at 0.14%. Aortic lesions were not present in any of the animals.

Topics: Animals; Aorta; Cholesterol; Chromatography, Gas; Diet; Esters; Male; Phytosterols; Rabbits; Sitosterols; Weight Gain

Cholesterol and other sterols exit the body primarily by secretion into bile. In patients with sitosterolemia, mutations in either of two ATP-binding cassette (ABC) half-transporters, ABCG5 or ABCG8, lead to reduced secretion of sterols into bile, implicating these transporters in this process. To elucidate the roles of ABCG5 and ABCG8 in the trafficking of sterols, we disrupted Abcg5 and Abcg8 in mice (G5G8(-/-)). The G5G8(-/-) mice had a 2- to 3-fold increase in the fractional absorption of dietary plant sterols, which was associated with an approximately 30-fold increase in plasma sitosterol. Biliary cholesterol concentrations were extremely low in the G5G8(-/-) mice when compared with wild-type animals (mean = 0.4 vs. 5.5 micromol ml) and increased only modestly with cholesterol feeding. Plasma and liver cholesterol levels were reduced by 50% in the chow-fed G5G8(-/-) mice and increased 2.4- and 18-fold, respectively, after cholesterol feeding. These data indicate that ABCG5 and ABCG8 are required for efficient secretion of cholesterol into bile and that disruption of these genes increases dramatically the responsiveness of plasma and hepatic cholesterol levels to changes in dietary cholesterol content.

Topics: Animal Feed; Animals; ATP Binding Cassette Transporter, Subfamily G, Member 5; ATP Binding Cassette Transporter, Subfamily G, Member 8; ATP-Binding Cassette Transporters; Bile; Biological Transport; Chimera; Cholestanol; Cholesterol; Cholesterol, Dietary; Crosses, Genetic; Dietary Fats; Female; Gene Targeting; Intestinal Absorption; Intestinal Mucosa; Lipoproteins; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Phytosterols; Sitosterols

The purpose of this project was to assess the plasma pharmacokinetics of [(3)H]cholesterol following coadministration of a novel vegetable stanol mixture composed of sitostanol and campestanol (FCP-3P4) to fasting rats. Following an overnight fast (12-16 h) and 48 h post-surgery, adult male Sprague Dawley rats were divided into six treatment groups and received a single-dose oral gavage at 0700 h of either: [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (5 mg/kg) + [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (12.5 mg/kg) + [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (25 mg/kg) + [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (50 mg/kg) + [(3)H]cholesterol (25 microCi/mL), or FCP-3P4 (100 mg/kg) + [(3)H]cholesterol (25 microCi/mL). Intralipid (10%) was the vehicle used to solubilize and coadminister [(3)H]cholesterol and FCP-3P4. Liquid chromatography-mass spectrometry analysis confirmed minimal cholesterol and vegetable stanol content within 10% Intralipid. Analysis of plasma pharmacokinetics was initiated by sampling 0.5 mL of blood prior to and 0.25, 0.5 1.0, 2.0, 4.0, 6.0, 8.0, 10, 24, 28, 32, and 48 h post-oral gavage. Plasma samples were obtained by centrifugation of the blood samples and analyzed for [(3)H]cholesterol radioactivity. Pharmacokinetics analysis was performed by standard noncompartmental methods using statistical moment theory. Thin-layer chromatography was used to confirm that the majority of radioactivity measured in plasma was cholesterol (in the form of esterified or unesterified cholesterol). Greater than 90% of the radioactivity measured in all plasma samples was cholesterol-associated (in the form of either esterified or unesterified cholesterol). The coadministration of FCP-3P4 significantly decreased the area under the curve of [(3)H]cholesterol concentration versus time from 0 to 48 h (AUC(0-48h)) and maximum concentration (C(max)) in a dose-dependent manner. However, coadministration of FCP-3P4 at 25, 50, and 100 mg/kg resulted in a significant increase in apparent total body clearance (CL/F, where F is the bioavailability constant), apparent volume of distribution (V(d)/F), and oral absorption rate constant (k(a)) of [(3)H]cholesterol compared with controls. These findings suggest that the novel vegetable stanol mixture, FCP-3P4, modifies the plasma pharmacokinetics of [(3)H]cholesterol in fasting rats on oral coadministration.

Topics: Administration, Oral; Animals; Anticholesteremic Agents; Cholesterol; Drug Interactions; Fasting; Intestinal Absorption; Male; Phytosterols; Rats; Rats, Sprague-Dawley; Sitosterols; Tritium

Plant sterols in vegetable foods might prevent colorectal cancer.. The objective was to study plant sterol intakes in relation to colorectal cancer risk in an epidemiologic study.. The study was performed within the framework of the Netherlands Cohort Study on Diet and Cancer in 120852 subjects who completed a baseline questionnaire in 1986. After 6.3 y of follow-up, 620 colon and 344 rectal cancer cases were detected. A case-cohort approach was used to calculate confounder-adjusted rate ratios (RRs) and their 95% CIs for quintiles of plant sterol intake.. The total mean (+/-SD) intake of campesterol, stigmasterol, beta-sitosterol, campestanol, and beta-sitostanol was 285 +/- 97 mg/d. Major contributors to plant sterol intake were bread (38%), vegetable fats (26%), and fruit and vegetables (21%). For men, there was no clear association between intake of any of the plant sterols and colon cancer risk when age, smoking, alcohol use, family history of colorectal cancer, education level, and cholecystectomy were controlled for. Adjustment for energy did not alter the result. For rectal cancer, adjustment for energy resulted in positive associations between risk and campesterol and stigmasterol intakes. For women, there was no clear association between intake of any of the plant sterols and colorectal cancer risk.. A high dietary intake of plant sterols was not associated with a lower risk of colon and rectal cancers in the Netherlands Cohort Study on Diet and Cancer.

Topics: Aged; Bread; Case-Control Studies; Cholesterol; Cohort Studies; Colorectal Neoplasms; Confounding Factors, Epidemiologic; Dietary Fats; Female; Follow-Up Studies; Fruit; Humans; Hypolipidemic Agents; Male; Middle Aged; Netherlands; Phytosterols; Prospective Studies; Rectal Neoplasms; Risk Factors; Sitosterols; Stigmasterol; Surveys and Questionnaires; Vegetables

The aim of this study was to study the inhibitory effect of dietary stanols (campestanol and sitostanol) fatty acid esters (SE) on intestinal cholesterol absorption. New Zealand white rabbits were fed regular chow alone or enriched with 0.2% cholesterol, 0.33% SE + cholesterol, 0.66% SE + cholesterol, 1.2% SE + cholesterol, 2.4% SE + cholesterol, and 1.2% SE alone. After 2 weeks, plasma cholesterol levels increased 3.6 times in the cholesterol group and did not decrease after addition of 0.33% or 0.66% SE to the cholesterol-enriched diets. However, after addition of 1.2% SE to the cholesterol diet, plasma cholesterol concentration decreased 50% (P <.001), but it did not decrease further after doubling of SE to 2.4%. Percent cholesterol absorption measured by the plasma dual-isotope ratio method was 73.0% +/- 8.1 % in the cholesterol group, which was similar to untreated baseline control. The percent absorption of cholesterol did not decrease significantly after addition of 0.33% or 0.66% SE to the cholesterol diet but decreased 43.8% (P <.001) in the 1.2% SE + cholesterol group, a finding similar to those in rabbits fed 1.2% SE alone. Increasing SE to 2.4% in the cholesterol diet did not further decrease absorption. Hepatic hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase activity reflecting cholesterol synthesis and low-density lipoprotein receptor-mediated binding unexpectedly decreased 67% (P <.01) and 57% (P <.05) in rabbits fed 1.2% SE alone. Increasing dietary SE intake to 1.2% reduced cholesterol absorption and plasma levels. Dietary SE intake below 1.2% was ineffective and above 2.4% did not further decrease percent absorption or plasma cholesterol levels. These results support the hypothesis that dietary SEs competitively displace cholesterol from intestinal micelles to reduce cholesterol absorption and decrease plasma cholesterol levels.

Topics: Animals; Anticholesteremic Agents; Bile; Cholestanetriol 26-Monooxygenase; Cholesterol; Cholesterol 7-alpha-Hydroxylase; Cholesterol, Dietary; Cytochrome P-450 Enzyme System; Dietary Supplements; Dose-Response Relationship, Drug; Enzyme Activation; Hydroxymethylglutaryl CoA Reductases; Intestinal Absorption; Liver; Male; Phytosterols; Rabbits; Receptors, LDL; Sitosterols; Steroid Hydroxylases

Phytosterols and phytostanols are known to lower low-density lipoprotein-cholesterol (LDL-C) levels in humans by up to 15%, and at least two products, Benecol and Take Control, are now on the market as naturally derived fatty acid esters of phytostanols (stanol esters) and phytosterols (sterol esters), respectively. A synthetic process was developed to synthesize gram quantities of trans-feruloyl-beta-sitostanol from ferulic acid and beta-sitostanol, with high purity and yields of approximately 60%. The process involves (a) condensation of trans-4-O-acetylferulic acid with the appropriate phytostanol or phytostanol mixture in the presence of N,N-dicyclohexylcarbodiimide and 4-(dimethylamino)pyridine, (b) separation of the trans-4-O-acetylferuloyl products by preparative liquid chromatography, (c) selective deacetylation of the feruloyl acetate, and (d) chromatographic purification of the feruloylated phytostanols. The process was successfully applied to synthesize stanol trans-feruloyl esters from "Vegetable Stanols", a mixture of approximately 70:30 beta-sitostanol and beta-campestanol, in comparable purity and yield.

Topics: Anticholesteremic Agents; Antioxidants; Coumaric Acids; Phytosterols; Sitosterols

As part of an extensive safety evaluation programme, a series of studies has been conducted to determine the fate of phytosterols in the rat. Rats were dosed by oral gavage with 14C-labelled samples of cholesterol, beta-sitosterol or beta-sitostanol or (3)H-labelled samples of beta-sitostanol, campesterol, campestanol or stigmasterol dissolved in sunflower seed oil. Urine and faeces were collected for up to 96 hours after dosing. There was no quantification of biliary excreted material in these studies. Animals were sacrificed and either prepared for whole body autoradiography or tissues and carcass remains were assayed for 14C or (3)H. The overall absorption of phytosterols was low as judged by tissue and carcass levels of radioactivity. Elimination from the body was mainly in the faeces and was initially very rapid, but traces of material were still being excreted at 4 days after dosing. While total absorption of the phytosterols could not be fully quantified without biliary excretion data, it was clear that cholesterol was absorbed to the greatest extent (27% of the dose in females at 24 hours). Campesterol (13%) was absorbed more than beta-sitosterol and stigmasterol (both 4%) which were absorbed more than beta-sitostanol and campestanol (1-2%). The absorption of phytosterols was slightly greater in females than males. For each test material, the overall pattern of tissue distribution of radioactivity was similar, with the adrenal glands, ovaries and intestinal epithelia showing the highest levels and the longest retention of radioactivity.

Topics: Animals; Autoradiography; Cholesterol; Female; Intestinal Absorption; Male; Phytosterols; Rats; Sitosterols; Stigmasterol; Tissue Distribution

We measured the percent absorption, turnover, and distribution of campestanol (24-methyl-5alpha-cholestan-3beta-ol) in a sitosterolemic homozygote, her obligate heterozygous mother, and three healthy human control subjects. For reasons relating to sterol hyperabsorption, the homozygote consumed a diet low in plant sterols that contained campestanol at about 2 mg/day. The heterozygote and three control subjects were fed a diet supplemented with a spread that contained campestanol at 540 mg/day and sitostanol (24-ethyl-5alpha-cholestan-3beta-ol) at 1.9 g/day as fatty acid esters. Plasma campestanol concentrations determined by capillary gas-liquid chromatography were 0.72 +/- 0.03 mg/dl in the homozygote, 0.09 +/- 0.04 mg/dl in the heterozygote, and 0.05 +/- 0.03 mg/dl for the control mean. After simultaneous pulse labeling with [3alpha-(3)H]campestanol intravenously and [23-(14)C]campestanol orally, the maximum percent absorption measured by the plasma dual-isotope ratio method as a single time point was 80% in the homozygote, 14.3% in the heterozygote, and 5.5 +/- 4.3% as the mean for three control subjects. Turnover (pool size) values estimated by mathematical analysis of the specific activity versus time [3alpha-(3)H]campestanol decay curves were as follows: 261 mg in the homozygote, 27.3 mg in the heterozygote, and 12.8 +/- 7.6 mg in the three control subjects (homogygote vs. controls, P < 0.001). The calculated production rate (mg/24 h) equivalent to actual absorption in the presence of dietary sterols and stanols was 0.67 mg/day or 31% of intake in the homozygote, 2.1 mg/day or 0.3% of intake in the heterozygote, and 0.7 +/- 0.3 mg/day or 0.1% of intake in the three control subjects. However, the excretion constant from pool A (K(A)) was prolonged markedly in the homozygote, but was 100 times more rapid in the heterozygote and three control subjects.Thus, campestanol, like other noncholesterol sterols, is hyperabsorbed and retained in sitosterolemic homozygotes. However, campestanol absorption was only slightly increased in the sitosterolemic heterozygote and removal was as rapid as in control subjects.

Topics: Adolescent; Adult; Carbon Radioisotopes; Cholesterol; Diet; Female; Half-Life; Heterozygote; Homozygote; Humans; Intestinal Absorption; Kinetics; Lipid Metabolism, Inborn Errors; Male; Middle Aged; Phytosterols; Sitosterols; Tritium

Campestanol (24-methyl-5alpha-cholestan-3beta-ol) is a naturally occurring plant stanol, structurally similar to cholesterol (5-cholesten-3beta-ol) and widely distributed in vegetable oils consumed in human diets. We measured the absorption and turnover of campestanol by the plasma dual-isotope ratio method and mathematical analysis of specific activity versus time decay curves after simultaneous oral and intravenous pulse-labeling with [3alpha-3H]- and [23-14C]-labeled campestanol, respectively, in New Zealand White (NZW) rabbits: six fed chow and six fed chow with 125 mg/d campestanol and 175 mg/d sitostanol (24-ethyl-5alpha-cholestan-3beta-ol). Plasma concentrations increased insignificantly from 0.08+/-0.01 to 0.09+/-0.01 mg/dL with dietary stanols. The percent campestanol absorption measured by the plasma dual-isotope ratio method after the rabbits were fasted for 6 hours yielded the percent absorption in the absence of competing intestinal sterols and stanols and declined insignificantly from 11.6%+/-3.5% in controls to 8.1%+/-3.7% in the treated rabbit groups. In contrast, the turnover, which measured actual absorption averaged over 24 hours, increased from 0.12+/-0.05 to 0.37+/-0.05 mg/d (P < .05) with campestanol and sitostanol added to the diet. However, the actual percent absorption declined from 3% to 0.3% of dietary intake with the campestanol and sitostanol-enriched diet. Campestanol pool sizes, although remaining small, increased slightly from 1.1+/-0.4 to 2.5+/-1.5 mg. The removal constant (KA) from pool A (MA) did not change significantly with added dietary campestanol and sitostanol (KA= -0.040+/-0.005 v -0.037+/-0.007 d(-1)). The results demonstrate small campestanol plasma concentrations and body pools even when the rabbits consumed substantial amounts because (1) intestinal absorption was limited and (2) was further reduced by competing dietary sitostanol, and (3) campestanol was removed rapidly from the body. Thus, campestanol, which shares the same basic structure and intestinal absorption pathway with cholesterol, does not accumulate when fed, and may be incorporated into the diet to block cholesterol absorption.

Topics: Administration, Oral; Animals; Anticholesteremic Agents; Biotransformation; Diet; Humans; Injections, Intravenous; Intestinal Absorption; Phytosterols; Rabbits; Sitosterols; Tissue Distribution

The aim of this study was to examine the efficacy of variable dietary sitostanol (SI) concentrations on cholesterol absorption, synthesis and excretion rates in two animal models. Hamsters and rabbits were fed semi-purified diets supplemented with cholesterol and 1% (w/w) phytosterols containing either 0.007, 0.17, 0.8 or 1% (w/w) SI. The control (0% (w/w) SI) groups consumed the same diets but no phytosterols were added. The dual-isotope plasma ratio of [13C]- and [18O]cholesterol and deuterium incorporation methods were applied to measure simultaneously cholesterol absorption and fractional synthesis, respectively. Plasma total cholesterol levels were lower in rabbits and hamsters fed 0.8 and 1% (w/w) SI, respectively, as compared to their controls. Percent cholesterol absorption was lower (P = 0.03) in hamsters fed 1% (w/w) SI (42.5 +/- 13.3%) than control (65.1 +/- 13.4%). Moreover, cholesterol excretion in the feces was 77 and 57% higher (P = 0.017) in the 1% (w/w) SI- relative to control- and 0.17% (w/w) SI-fed groups, respectively. In rabbits, cholesterol excretion was 64% higher (P = 0.018) in 0.8% (w/w) SI- compared with control-fed groups. Fractional synthesis rate was higher (P = 0.033) in hamsters fed 1% (w/w) SI (0.116 +/- 0.054 pool day(-1)) as compared to control (0.053 +/- 0.034 pool day(-1)). However, cholesterol synthesis rates did not vary among groups fed variable concentrations of SI. In rabbits, percent cholesterol absorption and its fractional synthesis rate varied but did not reach significance. Fractional synthesis rate in hamsters was correlated (r = -0.32, P = 0.03) with percent cholesterol absorption. In conclusion, dietary SI exhibited a dose-dependent action in inhibiting cholesterol absorption while increasing cholesterol excretion and upregulating cholesterogenesis in hamsters resulting in lower circulating lipid levels.

Topics: Analysis of Variance; Animals; Anticholesteremic Agents; Cholesterol; Cholesterol, Dietary; Cricetinae; Disease Models, Animal; Dose-Response Relationship, Drug; Feces; Lipids; Male; Phytosterols; Rabbits; Reference Values; Sitosterols; Species Specificity

Topics: Administration, Oral; Anticholesteremic Agents; Cross-Over Studies; Double-Blind Method; Drug Administration Schedule; Female; Humans; Male; Margarine; Phytosterols; Randomized Controlled Trials as Topic; Sitosterols

Total lipid extraction, solid-phase extraction, saponification, derivatization to trimethylsilyl ether derivatives, then capillary gas-liquid chromatography were used for quantitative analysis of sitosterol, campesterol, stigmasterol, sitostanol, campestanol, lathosterol, desmosterol, and lanosterol in human serum. Details of quality control integral to the accuracy and precision of analyses are included. The method limits of detection and quantitation, respectively, ranged from 0.05 microg/ml and 0.2 microg/ml for sitostanol to 0.4 microg/ml and 1.2 microg/ml for campesterol and campestanol. Analytes were measured at concentrations of 120 ng/ml to 6 microg/ml with standard deviations of 0.02 to 0.12 microg/ml for 55 analyses of a control serum sample conducted over a 2-month period.

Topics: Cholesterol; Chromatography, Gas; Desmosterol; Humans; Phytosterols; Quality Control; Reference Standards; Sitosterols; Time Factors

To examine how variable sitostanol (SI) levels in phytosterol-supplemented diets influence plasma and hepatic lipid concentrations, fifty hamsters were divided into five groups and fed semipurified diets containing 0.25% (w/w) cholesterol for 45 days ad libitum. Four groups were fed this diet with 1% (w/w) phytosterol mixtures which contained 0.01% (w/w) SI derived from soybean, 0.2% (w/w) SI derived from tall oil, 0.2% (w/w) synthetic SI mixture (SIM) and 1% (w/w) pure SI, respectively. A control group did not receive phytosterols. Dietary SI supplementation at 1% (w/w) decreased total and non-apolipoprotein-A cholesterol levels in plasma by 34% (P=0.001) and 55% (P=0.04), respectively, whereas mean plasma total cholesterol level in the 0.2% (w/w) SI group was 23% (P=0.001) lower than that of the control group. Conversely, plasma lipid profile in hamsters fed 1 or 0.2% (w/w) SI did not differ from the 0.01% (w/w) SI group. Liver weights were 15 and 20% (P=0.012) higher in the control group compared with those fed 0.01% and 1% (w/w) SI, respectively, while the hepatic cholesterol content in the control group was greater (P<0.0001) than that of all other groups. Plasma campesterol levels were higher (P=0.04) in the 0.01% and 0.2% (w/w) SI fed groups than in the control, 0.2% (w/w) SIM and 1% (w/w) SI groups. Hepatic sitosterol content was elevated (P=0.002) in the SIM fed group compared to other groups. We conclude that dietary SI effect is proportional to its concentration in phytosterol mixtures and in the diet. Dietary SI lowered plasma cholesterol levels at concentrations higher than 0.2% (w/w) in hamsters. (c) 1998 Elsevier Science B.V.

Topics: Animals; Anticholesteremic Agents; Body Weight; Cholesterol; Cholesterol, HDL; Cricetinae; Diet; Eating; Lipids; Liver; Organ Size; Phytosterols; Plant Oils; Sitosterols

The effects of graded amounts of dietary sitostanol (0.01, 0.2 and 0.8% (w/w)) were examined on plasma lipid-profile, coronary artery plaque development and lecithin:cholesterol acyl transferase activity in male New Zealand White rabbits given semi-purified diets for 10 weeks. All diets provided < 10% energy in the form of fat and contained 0.5% (w/w) cholesterol (C). Rabbits fed the semi-purified diet with 0.8% (w/w) (0.64 g/day) sitostanol had lower plasma total cholesterol (TC) (p = 0.006) (15.2 +/- 4.80 mmol/l) and very low-density lipoprotein-cholesterol (VLDL-C) (p = 0.007) (6.31 +/- 3.11 mmol/l) levels compared to the atherogenic control group (n = 6) (29.6 +/- 5.52 and 17.16 +/- 7.43 mmol/l, respectively). Dietary sitostanol at 0.8% (w/w) depressed plaque accretion in coronary arteries (p = 0.0014) and ascending aorta (p = 0.0004) compared with the atherogenic control, 0.01 and 0.2% (w/w) sitostanol-fed groups. No differences (p = 0.24) in the activity of lecithin:cholesterol acyl transferase (LCAT) were observed across groups, although plasma cholesterol fractional esterification rate was higher (p = 0.004) in the 0.8% (w/w) sitostanol fed animals compared with the atherogenic control. Significant negative correlations were demonstrated between sitostanol intake and plasma TC, LDL-C and VLDL-C levels. Hepatic campesterol levels were correlated (r = 0.3, p = 0.03) with plasma but not hepatic TC concentrations. These results demonstrate that dietary sitostanol at a concentration of 0.8% (w/w) or 0.64 g/day lowered plasma cholesterol levels and depressed atherosclerosis development in rabbits, but did not alter LCAT activity.

Topics: Animals; Anticholesteremic Agents; Arteriosclerosis; Cholesterol; Coronary Vessels; Diet; Esterification; Male; Phosphatidylcholine-Sterol O-Acyltransferase; Phytosterols; Rabbits; Sitosterols

Unesterified cholesterol (UC) that is taken up by the liver from lipoproteins is rapidly mixed by exchange with liver UC. Thus, it is not possible to quantitate the transport of UC from different lipoproteins into bile using radiolabeled UC. However, plant sterols do not exchange with UC and are secreted in bile with the same kinetics as UC. To compare the contribution to bile of sterols from different lipoproteins, we perfused isolated rat livers with VLDL, LDL, and HDL that were obtained from patients with hereditary phytosterolemia and were rich in plant sterols. After 30-min recirculating perfusions, hepatic concentrations of plant sterols were not different after different lipoproteins were perfused. However, biliary plant sterol secretion was markedly different: with the perfusion of either VLDL or LDL there was no increase in plant sterols in bile, but with perfusion of HDL, the secretion of plant sterols was increased two- to threefold (P = 0.0005). The increase in biliary plant sterols was detected 5-10 min after HDL was added to perfusates and was similarly large for each of three individual plant sterols that was tracked. Results show that when sterol transport from lipoproteins into bile can be determined, only HDL provides a vehicle for UC elimination in bile that is consistent with its putative function in reverse cholesterol transport.

Topics: Animals; Bile; Biological Transport; Cholesterol; Chromatography, High Pressure Liquid; Humans; Hypolipoproteinemias; Lipoproteins, HDL; Lipoproteins, LDL; Lipoproteins, VLDL; Liver; Male; Perfusion; Phytosterols; Rats; Rats, Sprague-Dawley; Sitosterols

Absorption of dietary cholesterol, campesterol, and sitosterol, cholesterol balance, and fecal excretion of plant sterols were determined in three unrelated patients with phytosterolemia and three healthy volunteers during constant intake of cholesterol and plant sterols using accurate gas-liquid chromatography-mass spectrometry techniques. Each subject received a mixture of [26,26,26,27,27,27-2H6]cholesterol, [6,7,7-2H3]sitostanol, and [6,7,7-2H3]campesterol together with two non-absorbable markers, [5,6,22,23-2H4]sitostanol and chromic oxide. Feces were collected from days 5 to 7 and absorption of different sterols was calculated from the intestinal disappearance of the different sterols relative to [5,6,22,23-2H4]sitostanol and chromic oxide. The results obtained by the two markers were not different and the absorption of cholesterol averaged 53 +/- 4% for the patients (mean +/- SD) and 43 +/- 3% for the volunteers. Campesterol absorption averaged 24 +/- 4% in patients and 16 +/- 3% in healthy volunteers, whereas sitosterol absorption averaged 16 +/- 1% and 5 +/- 1%, respectively. Cholesterol synthesis expressed by body weight varied considerably in the two groups but appeared to be about 5 times lower in patients than in controls. Administration of a high dose of sitostanol (0.5 g t.i.d.) to two patients was followed by a reduction in cholesterol absorption by 24% and 44%, an increase in fecal output of cholesterol and steroids derived from cholesterol and plant steroids, and a marked reduction of serum cholesterol, campesterol, and sitosterol. Under the conditions used, inhibition of cholesterol absorption by sitostanol was not followed by a significant rise in cholesterol synthesis. The time of observation was, however, too short to allow final conclusion on this. The results show that the absolute difference in absorption rate of different sterols between the patients and healthy volunteers was about the same. As a consequence, increasing hydrophobicity causes a relative decrease of absorption rates. Thus, patients with phytosterolemia seem to have a generally increased absorption of sterols rather than a loss of a specific discriminatory mechanism, and oral administration of sitostanol seems to be an interesting new approach for treatment of phytosterolemia.

Topics: Absorption; Adult; Anticholesteremic Agents; Bile Acids and Salts; Cholesterol; Cholesterol, Dietary; Deuterium; Dietary Fats; Feces; Female; Humans; Intestinal Mucosa; Lipid Metabolism, Inborn Errors; Lipoproteins; Male; Middle Aged; Phytosterols; Sitosterols; Steroids; Sterols

To investigate the action and mechanism of a dietary phytosterol mixture naturally containing sitostanol, derived from tall-oil, on circulating cholesterol and lipoprotein levels, five groups of rats were fed a control elemental diet (group 1), a control elemental diet with 1% cholesterol alone (group 2) or with sitostanol mixtures or a sitostanol-free mixture supplemented at 0.2% (group 3), 0.5% (group 4) or 1% (group 5) of dietary levels. One per cent supplementation of sitostanol (21%) compared with sitostanol-free mixtures decreased (P < 0.02) total serum cholesterol. Dietary sitostanol (16% or 21%) mixture at 1% dietary levels decreased (P < 0.05) low density lipoprotein (LDL) cholesterol and increased (P < 0.05) high density lipoprotein (HDL) cholesterol levels. The decrease of LDL and increase of HDL cholesterol were correlated (P < 0.01) with the level of sitostanol mixture in the diet. Consumption of the sitostanol-containing mixture (1% dietary levels) caused a compensatory increase in cholesterol synthesis as indicated by elevated (P < 0.05) lathosterol/ cholesterol ratios in plasma and hepatic cholesterol fractional synthesis rate (FSR) (P < 0.02). Both sitostanol and sitostanol-free mixtures at 0.5% or 1% dietary intake levels increased plasma campesterol and beta-sitosterol levels, while plasma sitostanol levels were negligible. The absence of sitostanol in plasma and the increase in cholesterol synthesis induced by dietary sitostanol mixtures in addition to elevation of plasma campesterol and beta-sitosterol by sitostanol or sitostanol-free mixtures suggest that sitostanol mixtures effectively modify circulating lipoprotein cholesterol concentrations at the level of the intestine, rather than internally at the level of cholesterogenesis.

Topics: Animals; Anticholesteremic Agents; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Dietary Fats, Unsaturated; Drug Interactions; Eating; Glycine max; Liver; Male; Phytosterols; Plant Oils; Rats; Rats, Wistar; Sitosterols; Structure-Activity Relationship

This study was undertaken to compare the ability of two plant sterols to reduce serum levels of lipids and to compare their mechanism of action in nine children with severe familial hypercholesterolemia (total and low-density lipoprotein cholesterol concentrations averaged 9.57 mmol/L (370 mg/dl) and 7.87 mmol/L (301 mg/dl)). After a 3-month strict diet, the children were given sitosterol pastils (2 gm three times a day) for 3 months, followed by a 7-month course of sitostanol (0.5 gm three times a day). Serum lipoprotein levels and serum concentrations of campesterol and sitosterol were determined in all nine children, and the fecal excretion of neutral and acidic sterols were determined in seven children at the end of each therapeutic regimen. Sitosterol reduced low-density lipoprotein cholesterol levels by 20% (p < 0.01); sitostanol reduced low-density lipoprotein cholesterol levels by 33% after 3 months and 29% after 7 months (p < 0.01 compared with diet; p < 0.05 compared with sitosterol). Although sitosterol did not alter serum concentrations of campesterol and sitosterol, a significant reduction did occur during sitostanol therapy (-47% and -51%, respectively; p < 0.01). Fecal excretion of neutral sterols increased from 6.7 mg/kg per day during the control period to 9.7 mg/kg per day during sitosterol administration (p < 0.05), and to 12.6 mg/kg per day during sitostanol administration (p < 0.05 compared with diet and sitosterol periods), indicating an increase in the inhibition of intestinal cholesterol absorption. All children completed the study and no obvious side effects occurred. The data indicate that sitostanol, even with a dose four-fold lower than that of sitosterol, was significantly more effective in reducing elevated levels of low-density lipoprotein cholesterol, and the reduction in serum lipid levels was of the same magnitude as that observed with systemic lipid-lowering drugs. These results suggest that sitostanol, a nonabsorbable plant sterol, could be the drug of choice for treating familial hypercholesterolemia in childhood.

Topics: Adolescent; Alanine Transaminase; Alkaline Phosphatase; Apolipoproteins B; Carotenoids; Child; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Feces; Female; Heterozygote; Humans; Hyperlipoproteinemia Type II; Male; Phytosterols; Sitosterols; Sterols

Rapeseed oil fed to 24 hypercholesterolemic patients (50 g/day) reduced serum cholesterol (-8.5%) and cholestanol concentrations but increased those of campesterol and sitosterol. Continuation of rapeseed oil alone or with added sitosterol (625 mg/day) or sitostanol (630 mg/day) had no further effect on serum cholesterol. Rapeseed oil with sitosterol increased further its own proportion to cholesterol in serum but reduced that of campesterol while rapeseed oil with sitostanol reduced the proportions of both sitosterol and campesterol proportionately to the pretreatment values. The changes in the campesterol and sitosterol proportions were negatively and positively related to each other during the sitosterol and sitostanol additions, respectively. Thus, concentrations of unsaturated plant sterols in serum reflect their dietary intakes, saturated plant sterols are virtually not absorbed, plant sterols interfere with absorption of unsaturated structurally different plant sterols and cholestanol, and plant sterol-induced reduction of sterol absorption may be positively related to absorption efficiency of sterols.

Topics: Adult; Body Weight; Brassica; Cholesterol; Dietary Fats; Female; Humans; Hypercholesterolemia; Male; Middle Aged; Phytosterols; Plant Oils; Sitosterols