Influence of fatty acid patterns on the intestinal absorption pathway of quercetin in thoracic lymph duct-cannulated rats.

Since it is known that dietary fats improve the bioavailability of the flavonol quercetin, we purposed to investigate whether this effect is due to increased lymphatic transport of quercetin. In rats with implanted catheters in the thoracic lymph duct, we administered quercetin into the duodenum with TAG emulsions containing either long-chain fatty acids (LCT) or medium-chain fatty acids (MCT). Controls received quercetin together with a glucose solution. LCT administration increased the lymphatic output of quercetin (19.1 (SEM 1.2) nmol/8 h) as well as the lymph-independent bioavailability of the flavonol, determined as area under the plasma concentration curve (1091 (SEM 142) microM x min). Compared with glucose administration, MCT neither increased the lymphatic output (12.3 (SEM 1.5) nmol/8 h) nor the bioavailability of quercetin (772 (SEM 99) microM x min) significantly (glucose group: 9.8 (SEM 1.5) nmol/8 h and 513 (SEM 55) microM x min, respectively). Because LCT are released within chylomicrons into the intestinal lymph while MCT are mainly released into the portal blood, we conclude from the present results that dietary fats that are mainly composed of LCT improve quercetin bioavailability by increasing its transport via the lymph, thereby circumventing hepatic first-pass metabolism of the flavonol. In addition, LCT could enhance quercetin absorption by improving its solubility in the intestinal tract.

Chronic quercetin feeding decreases plasma concentrations of salicylamide phase II metabolites in pigs following oral administration.

We investigated acute effects and effects after chronic intake of the orally administered flavonol quercetin on pharmacokinetics of salicylamide metabolites (SAM) after oral administration of salicylamide in pigs. Salicylamide (8 mg/kg body weight) was orally administered to seven pigs either without or with quercetin (10 mg/kg body weight). Additionally, salicylamide was administered to five pigs that had received a diet supplemented with the flavonol for 1 week. Daily quercetin intake was 10 mg/kg in these animals. Co-ingestion of quercetin with the drug did not alter area under the concentration-time curve (AUC(0→∞)), time to achieve maximum plasma concentration (t(max)), mean residence time (MRT) or half-life (t(1/2)) of SAM. However, maximum plasma concentration (c(max)) of SAM was lower when quercetin was administered concomitantly. After quercetin pre-treatment for 1 week AUC(0→∞), t(1/2) and MRT of SAM were decreased, while other parameters investigated were not affected. Co-ingestions and dietary pre-treatment with quercetin influenced SAM metabolism after oral salicylamide intake. But effects seen after acute concomitant intake are rather explained by induced salicylamide excretion from the intestinal mucosa, whereas quercetin pre-treatment seemed to induce hepatic enzymes involved in phase-II metabolism and thereby enhanced elimination of SAM.

Oral bioavailability of quercetin from different quercetin glycosides in dogs.

Although the flavonol quercetin is used as a supplement in commercial dog food, data on quercetin bioavailability in dogs are not available. Thus, we investigated quercetin bioavailability (measured as area under the concentration-time curve) in nine adult beagle dogs at an oral dose of 10 mg/kg body weight (b.w.). The major fraction (>80 %) of flavonols circulating in blood plasma were conjugated metabolites of quercetin. The absolute bioavailability of quercetin (i.e. the fraction that reaches the systemic circulation) was only about 4 %. We also compared the oral bioavailability between the aglycone quercetin and its more often used glucorhamnoside (rutin) and 3-O-glucoside (isoquercitrin) at an equimolar dose of 30 mumol/kg b.w. (corresponding to 10 mg quercetin/kg). Quercetin and isoquercitrin were mainly absorbed in the small intestine with isoquercitrin being one and a half times more bioavailable than quercetin. Maximal plasma concentration after isoquercitrin treatment was 0.89 (sem 0.07) mumol/l. Although quercetin absorption from rutin was delayed, relative bioavailability was not lower than from the aglycone itself. The latter observation is in clear contrast to findings in human subjects, pigs or rats and might indicate that rutin is a better source of quercetin in dogs than in other species. However, potential in vivo quercetin effects beyond the gastrointestinal tract are limited by the intensive metabolism as well as by the rather low bioavailability of this flavonol.

Gene Regulatory Effects of Ginkgo biloba Extract and Its Flavonol and Terpenelactone Fractions in Mouse Brain.

The standardised Ginkgo biloba extract EGb761 is known for its potential beneficial effects in the prevention and therapy of neurodegenerative disorders including Alzheimer's disease (AD). However, the molecular mechanisms and the specific role of its constituents are largely unknown. The aim of the present feeding trial was to investigate the effects of EGb761 and its major constituents on the expression of genes encoding for proteins involved in the pathogenesis of AD in mouse brain. Six month old C57B6 mice were fed semi synthetic diets enriched with either EGb761 or one of its main fractions, flavonols and terpenelactones, respectively, over a period of 4 weeks. Thereafter, mRNA of alpha-secretase, neprilysin, amyloid precursor protein (App), App binding protein-1 and acetylcholine esterase was quantified in hippocampus and cortex. EGb761 and its flavonol fraction had no effects on relative mRNA levels of the respective genes in mouse brain. However, the terpenelactone fraction significantly decreased the mRNA levels of App in the hippocampus. Taken together, a 4 week dietary treatment with EGb761 or its main fractions had only moderate effects on mRNA levels of AD related genes in cortex and hippocampus of mice.

Effects of the flavonol quercetin on the bioavailability of simvastatin in pigs.

The influence of the dietary flavonol quercetin on the pharmacokinetics of the HMG-CoA reductase inhibitor simvastatin was investigated in pigs. Simvastatin (0.25mg/kg body weight) was orally administered to six pigs either without or with quercetin (10mg/kg). In addition, simvastatin was administered to three pigs that had received a diet supplemented with the flavonol over a period of 1 week. Daily quercetin intake was 10mg/kg in these animals. Co-ingestion of quercetin with the statin did not alter area under the concentration time curve (AUC(0-->infinity)), time to achieve maximum plasma concentration (t(max)) or half-life (t(1/2)) of simvastatin. However, there was a trend towards a reduction of the maximum plasma concentration (C(max)) of simvastatin when quercetin was administered concomitantly (P=0.06). As compared to controls, AUC(0-->infinity) of simvastatin was significantly decreased after feeding the quercetin-supplemented diet for 1 week. The plasma ratio of simvastatin and its acid metabolite was neither altered by the concomitant quercetin ingestion nor by feeding of the flavonol over a period of 1 week. We conclude that chronic ingestion of high doses of the flavonol quercetin will decrease the bioavailability of simvastatin to a significant extent.

The influence of postharvest processing and storage of foodstuffs on the bioavailability of flavonoids and phenolic acids.

Postharvest processing and storage not only influence the content and composition of flavonoids and phenolic acids in foodstuffs, thereby altering the amount of potentially bioavailable bioactive compounds, but can also modify their chemical form. Moreover, due to the intensive metabolism during absorption, the metabolites circulating in blood differ from the parent compounds found in food. Thus, it is difficult to predict potential in vivo effects of phenolic compounds merely by their contents in foodstuffs. Their specific bioavailability needs to be determined. This review considers studies regarding the bioavailability of flavonoids and phenolic acids from foodstuffs that meet the following criteria: providing actual concentrations of flavonoids and phenolic acids in blood plasma, body tissues, or urine, comparing differently stored or processed foods (excluding studies that use supplements or pure substances), and considering the high interindividual variability by repeated measurements in the same individuals. Only a few studies meet all of these criteria. In conclusion, processing and storage of food can have either positive or negative effects on the bioavailability of flavonoids and phenolic acids because these treatments may not only change the content, but also the chemical form of these compounds.

Effect of a short- and long-term treatment with Ginkgo biloba extract on amyloid precursor protein levels in a transgenic mouse model relevant to Alzheimer's disease.

Several clinical trials have reported beneficial effects of the Ginkgo biloba extract EGb761 in the prevention and therapy of cognitive disorders including Alzheimer's disease (AD). The aim of the present long-term feeding trial was to study the impact of dietary EGb761 on Amyloid precursor protein (APP) metabolism in mice transgenic for human APP (Tg2576). Tg2576 mice were fed diets with and without EGb761 (300 mg/kg diet) for 1 and 16 months, respectively. Long-term treatment (16 months) with EGb761 significantly lowered human APP protein levels by approximately 50% as compared to controls in the cortex but not in the hippocampus. However, APP levels were not affected by EGb761 in young mice. Current data indicate that APP seems to be an important molecular target of EGb761 in relation to the duration of the Ginkgo biloba treatment and/or the age of the animals. Potential neuroprotective properties of EGb761 may be, at least partly, related to its APP lowering activity.

Tissue distribution of quercetin in pigs after long-term dietary supplementation.

Although the flavonol quercetin is intensively investigated, our knowledge about its bioavailability and possible target organs is far from being complete. The aim of this study was to check the potential of quercetin to accumulate in various tissues after long-term dietary treatment compared with a single treatment with flavonol. Pigs ingested either a single dose of quercetin aglycone (25 mg/kg body weight; Expt. 1) or received the flavonol twice a day at the same dose mixed into their regular meals (i.e 50 mg.kg(-1).d(-1)) for 4 wk (Expt. 2). In both experiments, we took plasma and tissue samples 90 min after the final meal and analyzed them using HPLC. Additionally, the specific activity of the enzyme beta-glucuronidase was measured in selected tissues. Higher flavonol concentrations than in plasma were found in only the liver (Expt. 1) or the intestinal wall and kidneys (Expt. 2). All tissues except blood plasma contained a variable amount of deconjugated quercetin in the range of 30-100% of total flavonols. However, the specific beta-glucuronidase activity was not correlated with the proportions of deconjugated flavonols in the various tissues. Long-term dietary intake of the flavonol did not lead to a greater accumulation in any tissue compared with the single treatment. Flavonol concentrations only exceeded the plasma concentration within organs involved in its metabolism and excretion, including liver, small intestine, and kidneys.

Effect of dietary flavonoids on pathways involved in drug metabolism.

Flavonoids are a large group of plant polyphenols with presumed beneficial effects on several common diseases. The use of dietary supplements enriched with flavonoids is becoming increasingly popular. These polyphenols are substrates of enzymes like cytochrome P450 monooxygenases and phase II conjugation enzymes, as well as of drug transporters involved in drug excretion. Thus, they share the same metabolic pathways with many therapeutic drugs. A number of studies have demonstrated inhibition of various cytochrome P450 monooxygenases and drug transporters by flavonoids. Flavonoid-induced effects on drug bioavailability were also shown. This raises concerns about the safe use of flavonoid supplements and flavonoid-containing remedies which are not subject to legal regulations. The challenge is to find a suitable way to predict harmful drug-flavonoid interactions.

Ginkgo biloba extract and its flavonol and terpenelactone fractions do not affect beta-secretase mRNA and enzyme activity levels in cultured neurons and in mice.

Numerous clinical trials have reported beneficial effects of the Ginkgo biloba extract EGb761 in the prevention and therapy of cognitive disorders including Alzheimer's disease (AD). Although neuroprotective properties of EGb761 have been consistently reported, the molecular mechanisms of EGb761 and the specific role of its major constituents, the flavonols and terpenlactones, are largely unknown. One major hallmark of AD is the deposition of amyloid-beta (A beta) as amyloid plaques in the brain. A beta is a cleavage product of amyloid precursor protein (APP). Certain proteases, called beta-secretases (BACE), are crucial in the formation of A beta. The purpose of the present study was to investigate the efficacy of EGb761 and its flavonol and terpenelactone fraction to modulate BACE-1 enzyme activity and mRNA levels in vitro and in vivo. Neither EGb761 nor its fractions affected BACE-1 activity in vitro. Furthermore, also in Neuro-2a cells and wild-type as well as transgenic (Tg2576) laboratory mice, no significant effect of EGb761 on BACE-1 enzyme activity and mRNA levels were observed. Current findings suggest that BACE-1 may not be a major molecular target of EGb761 and its flavonol and terpenelactone fraction.

The fatty acid pattern of dietary fat influences the oral bioavailability of the flavonol quercetin in pigs.

We have shown recently that dietary fat content influences the bioavailability of the flavonol quercetin. In the present study, the influence of the fatty acid pattern of dietary fats on the oral bioavailability of quercetin was investigated. Quercetin (30 micromol/kg body weight) was administered to growing pigs (n 6) in test meals consisting either of 200 g of a standard pig diet (2% crude fat) or of the same diet supplemented with 15 g fat/100 g diet using either medium-chain (MCT) or long-chain fatty acid triacylglycerols (LCT). Blood samples were drawn repeatedly over a period of 24 h and analysed by HPLC. In addition, the influence of the different diets on gastric emptying was investigated in rats. In pigs, the bioavailability of quercetin was measured by quantifying its plasma metabolites with an intact flavonol structure. Bioavailability was enhanced by 38% (P<0.05) and 12% (P>0.05) after intake with the MCT and LCT diets, respectively, compared to the standard diet. Maximum plasma concentrations of quercetin were reached significantly later with the MCT diet than with the LCT or the standard diet (P<0.05). No differences in dry matter of the gastric content were observed 60 min after intake of the experimental diets in rats. Thus, administration of quercetin together with a diet containing MCT fat enhances the bioavailability of the flavonol. Absorption of quercetin was delayed significantly with this diet. However, this was probably not due to slower gastric emptying of the MCT diet.

The potential of flavonoids to influence drug metabolism and pharmacokinetics by local gastrointestinal mechanisms.

In recent years, public and scientific interest in plant flavonoids has tremendously increased due to postulated health benefits. Whereas the amount of flavonoids ingested with the regular diet is rather low, the use of supplements enriched with these polyphenolics is becoming increasingly popular. This raises concerns about possible interactions of flavonoids with therapeutic drugs, because both are xenobiotics and, thus, share at least partially the same metabolic pathways. A number of in vitro studies have shown effects of flavonoids on enzymes involved in xenobiotic metabolism, like cytochrome P450 monooxygenases and phase II conjugation enzymes, or on membrane transporters involved in drug excretion. Several investigations have also reported changes of drug bioavailability by certain flavonoids. This article attempts to present an overview of flavonoid effects on pathways involved in drug metabolism. It focuses on phase I and phase II enzymes as well as on transporters involved in drug metabolism which are expressed in the gastrointestinal tract.

In vitro degradation of the flavonol quercetin and of quercetin glycosides in the porcine hindgut.

The present study investigated the microbial degradation of the plant flavonol quercetin and its naturally occurring glycosides isoquercitrin and rutin in the porcine hindgut. The experiments were carried out with the semicontinuous colon-simulation technique. The fluid and particle phase of pig hindgut contents from freshly slaughtered animals were used for the in vitro incubations. Following a five-day equilibration period, quercetin, isoquercitrin or rutin were administered to fermentation vessels and their turnover rate was determined. None of the flavonols affected parameters of microbial fermentation like pH, redox potential or VFA production. The turnover rate for isoquercitrin was seven times higher than the turnover for the fermentation fluid. The turnover rates for quercetin and rutin were four and twofold higher than fluid turnover, respectively. After administration of isoquercitrin or rutin, their aglycone quercetin was detected as an intermediary metabolite. Under sterile conditions using autoclaved incubation fluids and hindgut contents, turnover rates for quercetin and rutin were still higher than the fluid turnover in the fermentation vessels. This indicates a certain chemical instability of the flavonols and/or adsorption to ingesta particles. Thus, flavonols are subjected to microbial metabolism in the porcine hindgut. The glycosidic structure strongly influences the rate of metabolism.

Effect of the flavonol quercetin on membrane conductances in rat colonic crypt cells.

The plant polyphenol quercetin was shown to induce Cl- secretion in rat colon. This study was performed to investigate the alterations of membrane conductances in isolated epithelial cells induced by quercetin. Whole-cell patch-clamp recordings were performed in isolated crypts from rat distal colon. In cells of the crypt basis, quercetin significantly hyperpolarized the membrane potential at concentrations > or =3 microM and increased the K+ conductance without visibly altering the Cl- conductance. Thus, quercetin induces Cl- secretion merely by activation of K+ channels in the colon epithelium.

Quercetin glucosides inhibit glucose uptake into brush-border-membrane vesicles of porcine jejunum.

Recent experimental data point to an interaction of dietary flavonol monoglucosides with the intestinal Na-dependent glucose transporter 1 (SGLT1). To investigate this interaction in more detail, we performed experiments with SGLT1-containing brush-border-membrane vesicles (BBMV) from pig jejunum. The flavonol quercetin-3-O-glucoside (Q3G) concentration-dependently inhibited Na-dependent uptake of radioactively labelled d-glucose into BBMV. Uptake of l-leucine was not inhibited by Q3G, indicating a specific interaction of the glucoside with SGLT1. Whereas the maximal transport rate of concentration-dependent initial glucose uptake was not altered in the presence of Q3G, the constant for half-maximal glucose uptake was increased, suggesting a competitive type of inhibition of glucose uptake by Q3G. Trans-stimulation experiments suggested the transport of Q3G via SGLT1. In addition, Q3G decreased the Na-independent diffusive uptake of glucose into BBMV. Other flavonoids were also tested for their inhibitory effect on d-glucose uptake. Among the tested quercetin glycosides, only the 4'-O-glucoside (Q4G) also inhibited Na-dependent glucose uptake into BBMV, whereas the 3-O-galactoside, the 3-O-glucorhamnoside and the aglycone quercetin itself were ineffective. Glucosides of some other flavonoid classes such as naringenin-7-O-glucoside, genistein-7-O-glucoside and cyanidin-3,5-O-diglucoside were ineffective as well. Thus, dietary quercetin monoglucosides, for example, Q3G and Q4G, have an impact on intestinal nutrient transporters such as SGLT1 and related systems.

Bioavailability of quercetin in pigs is influenced by the dietary fat content.

The flavonol quercetin is one of the most prevalent flavonoids found in edible plants. In this study, the influence of dietary fat on oral bioavailability of quercetin was investigated. Quercetin (30 micromol/kg body weight) was administered either as the lipophilic aglycone or as the more hydrophilic quercetin-3-O-glucoside in test meals differing in fat content (3, 17, or 32 g fat/100 g diet) to growing pigs. Blood samples were drawn repeatedly over a 24-h period and analyzed by HPLC. The main metabolite found in plasma was always conjugated quercetin. Quercetin bioavailability from each diet was always higher from the glucoside than from the aglycone. Irrespective of the chemical form applied, the bioavailability of quercetin was higher in the 17% fat diet compared with the 3% fat diet (P < 0.05). No further effect on bioavailability was observed when the flavonols were administered with diets containing 32% fat. The elimination of quercetin was significantly delayed after its application with fat-enriched diets (P < 0.05). Thus, in addition to the chemical form of the flavonol, the fat content of the diet influences oral bioavailability of quercetin.

The bioavailability of quercetin in pigs depends on the glycoside moiety and on dietary factors.

Recent investigations suggest that the bioavailability of quercetin depends on the glycoside moiety of the quercetin glycosides present in the diet. In this study, we compared the oral bioavailability of quercetin from quercetin aglycone and two different quercetin glycosides in pigs. Pigs were equipped with permanent catheters in the jugular and portal veins. After consumption of a test meal containing the respective compounds, blood samples were drawn repeatedly over a period of 24 h and analyzed by HPLC. In a first set of experiments, pigs received a single oral dose of 148 micromol/kg body (equivalent to 50 mg/kg) provided as quercetin aglycone, quercetin-3-O-glucoside (Q3G) or quercetin-3-O-glucorhamnoside (rutin) as part of their diet. The main metabolite in plasma was always conjugated quercetin, whereas free quercetin was not detected in either the jugular or the portal blood. For Q3G and rutin, the relative total bioavailability of quercetin (i.e., conjugated quercetin and conjugated methylethers of quercetin) was 148% (P = 0.07) and 23% (P < 0.05), respectively, compared with quercetin aglycone. In another experiment with a dose of 29.6 micro mol/kg (equivalent to 10 mg/kg), the relative total bioavailability of Q3G was 167% compared with the aglycone (P < 0.05). Bioavailability of Q3G was significantly higher when the test meal was ground beef rather than the standard ration. Our results indicate that the bioavailability of quercetin from quercetin glycosides is determined by a complex interdependence between the chemical form of the flavonols and dietary factors.

Gossypol induces chloride secretion in rat proximal colon.

The effect of gossypol on electrolyte transport was investigated in rat colon mounted in Ussing chambers. The addition of gossypol to the mucosal or serosal side led to an increase in mucus secretion, which we did not quantify. Mucosally or serosally added gossypol also induced a rise in short circuit current (I(sc)) and tissue conductance (G(t)). Part of the mucosally added gossypol seemed to be bound to the mucus because the effects on I(sc) and G(t) were smaller when gossypol was added to the mucosal side. Serosally added gossypol had an effect on I(sc) at a concentration of 10 micromol l(-1). Mucus secretion was reduced in low Ca(2+) buffer. The increase in I(sc) was diminished by blockers of Cl- channels, K+ channels, of the Na+/K+ ATPase and of the Na+/K+/2 Cl- cotransporter. Measurements of unidirectional ion fluxes showed that gossypol added to the mucosal side had no effect on net Na+ transport, but increased Cl- secretion. The effect of mucosally added gossypol was significantly reduced by the use of low Cl- buffers and abolished when the buffer was additionally depleted of HCO(3)(-). Calmodulin antagonists inhibited the effect on secretion. These findings indicate that gossypol induces chloride secretion via a calmodulin-dependent mechanism. High concentrations of gossypol induced a strong increase in G(t) that could be blocked by W7, a blocker of calmodulin-dependent myosin light chain kinase. This indicates that the rise in G(t) is not due to an unspecific toxic effect, but instead, to specific opening of tight junctions.

The flavonol quercetin activates basolateral K(+) channels in rat distal colon epithelium.

1. The flavonol quercetin has been shown to activate a Cl(-) secretion in rat colon. Unlike the secretory activity of the related isoflavone genistein, quercetin's secretory activity does not depend on cyclic AMP; instead, it depends on Ca(2+). We investigated the possible involvement of Ca(2+) dependent basolateral K(+) channels using apically permeabilized rat distal colon epithelium mounted in Ussing chambers. 2. In intact epithelium, quercetin induced an increase in short-circuit current (I(sc)), which was diminished by the Cl(-) channel blockers NPPB and DPC, but not by glibenclamide, DIDS or anthracene-9-carboxylic acid. The effect of the flavonol was also inhibited by several serosally applied K(+) channel blockers (Ba(2+), quinine, clotrimazole, tetrapentylammonium, 293B), whereas other K(+) channel blockers failed to influence the quercetin-induced increase in I(sc) (tetraethylammonium, charybdotoxin). 3. The apical membrane was permeabilized by mucosal addition of nystatin and a serosally directed K(+) gradient was applied. The successful permeabilization was confirmed by experiments demonstrating the failure of bumetanide to inhibit the carbachol-induced current. 4. In apically permeabilized epithelium, quercetin induced a K(+) current (I(K)), which was neither influenced by ouabain nor by bumetanide. Whereas DPC, NPPB, charybdotoxin and 293B failed to inhibit this I(K), quinine, Ba(2+), clotrimazole and tetrapentylammonium were effective blockers of this current. 5. We conclude from these results that at least part of the quercetin-induced Cl(-) secretion can be explained by an activation of basolateral K(+) channels.