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Exploring and disentangling the production of potentially bioactive phenolic catabolites from dietary (poly)phenols, phenylalanine, tyrosine and catecholamines
Abstract
Following ingestion of fruits, vegetables and derived products, (poly)phenols that are not absorbed in the upper gastrointestinal tract pass to the colon, where they undergo microbiota-mediated ring fission resulting in the production of a diversity of low molecular weight phenolic catabolites, which appear in the circulatory system and are excreted in urine along with their phase II metabolites. There is increasing interest in these catabolites because of their potential bioactivity and their use as biomarkers of (poly)phenol intake. Investigating the fate of dietary (poly)phenolics in the colon has become confounded as a result of the recent realisation that many of the phenolics appearing in biofluids can also be derived from the aromatic amino acids, l-phenylalanine and l-tyrosine, and to a lesser extent catecholamines, in reactions that can be catalysed by both colonic microbiota and endogenous mammalian enzymes. The available evidence, albeit currently rather limited, indicates that substantial amounts of phenolic catabolites originate from phenylalanine and tyrosine, while somewhat smaller quantities are produced from dietary (poly)phenols. This review outlines information on this topic and assesses procedures that can be used to help distinguish between phenolics originating from dietary (poly)phenols, the two aromatic amino acids and catecholamines.
... These metabolites result from the metabolism of a wide variety of different phenolic compounds that occur in multiple dietary sources, including crops and forage [49,50]. They can also be produced endogenously from other substrates, including the aromatic amino acids phenylalanine and tyrosine [51]. ...
... For instance, in a recent study, the most discriminating compounds responsible for distinguishing milk samples based on animal feeding were hippuric acid, hydroxyhippuric acid, enterolactone, and isoflavone metabolites like equol and O-desmethylangolensin [61]. However, the use of hippuric acid in this context might be questionable in view of a recent publication by Clifford et al. [51]. In practice, determining the relationships among milk types, sterilization treatments, fat and lactose contents, and phenolic metabolite concentrations, is not straightforward. ...
Milk holds a high nutritional value and is associated with diverse health benefits. The understanding of its composition of (poly)phenolic metabolites is limited, which necessitates a comprehensive evaluation of the subject. This study aimed at analyzing the (poly)phenolic profile of commercial milk samples from cows and goats and investigating their sterilization treatments, fat content, and lactose content. Fingerprinting of phenolic metabolites was achieved by using ultra-high-performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UHPLC-QqQ-MS/MS). Two hundred and three potential microbial and phase II metabolites of the main dietary (poly)phenols were targeted. Twenty-five metabolites were identified, revealing a diverse array of phenolic metabolites in milk, including isoflavones and their microbial catabolites equol and O-desmethylangolensin, phenyl-γ-valerolactones (flavan-3-ol microbial catabolites), enterolignans, urolithins (ellagitannin microbial catabolites), benzene diols, and hippuric acid derivates. Goat’s milk contained higher concentrations of these metabolites than cow’s milk, while the sterilization process and milk composition (fat and lactose content) had minimal impact on the metabolite profiles. Thus, the consumption of goat’s milk might serve as a potential means to supplement bioactive phenolic metabolites, especially in individuals with limited production capacity. However, further research is needed to elucidate the potential health effects of milk-derived phenolics.
... This activity is a key factor in the browning of fruits and vegetables when they are damaged or cut. PPO binds to phenolic compounds such as catechol tyrosine [2][3][4] that is present in the plant tissues. The enzyme then catalyzes the oxidation of these phenolic compounds into quinones where oxygen is used as co-substrate. ...
Polyphenol oxidase(PPO) is a food enzyme which holds a very important place in the food industry mainly because of its impairing effects. This enzyme leads to the darkening of tissue when fruits are cut or any damage occurs which leads to the depreciation of nutritional value and food acceptability. It is not clear which roles this compound may have in cell metabolism. In plants, PPO contributes to defence mechanisms against pests and pathogens by producing toxic quinones that deter herbivory and inhibit microbial growth. In this report, the possible antifungal effects were examined. Analyses are carried out by properties of standard polyphenolic substances oxidized by PPO were assayed on Red delicious apple extract whose phenolics were previously isolated.
The metabolomics of Newhall navel oranges with geographical indication (GI) was measured by LC-MS/MS from three areas Ganzhou, Fengjie, and Zigui in China. Orthogonal partial least squares-discriminant analysis was performed for samples classification and important metabolites identification. This approach identified the best markers (i.e. tricarboxylic acid, amino acids, and phenolics) of the geographical origin able to discriminated Fengjie, Ganzhou, and Zigui organge samples. The results showed that peeled-samples from Zigui were richest and those from Fengjie lowest in all of the top ten most important markers. In whole samples, the highest levels of seven important markers occurred in those from Zigui, and the other three important markers appeared the highest levels in those from Ganzhou. According to the variation of the best markers, and the highest value of titratable acidity in those from Zigui, the samples planted in Ganzhou have the best balance between taste and nutrition. This work confirms the approach of metabolomics combined with OPLS-DA is an effective way for origin tracing and overall quality evaluation.
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Free or conjugated aromatic/phenolic acids arise from the diet, endogenous metabolism of catecholamines (adrenaline, noradrenaline, dopamine), protein (phenylalanine, tyrosine), pharmaceuticals (aspirin, metaprolol) plus gut microbiota metabolism of dietary (poly)phenols and undigested protein. Quantitative data obtained with authentic calibrants for 112 aromatic/phenolic acids including phase-2 conjugates in human plasma, urine, ileal fluid, feces and tissues have been collated and mean/median values compared with in vitro bioactivity data in cultured cells. Ca 30% of publications report bioactivity at ≤1 μmol/L. With support from clinical studies, it appears that the greatest benefit might be produced in vascular tissues by C6-C3 metabolites, including some of gut microbiota origin and some phase-2 conjugates, 15 of which are 3',4'-disubstituted with multiple sources including caffeic acid and hesperetin, plus one unsubstituted and two mono-substituted examples which can originate from protein. There is an unexamined potential for synergy. Free-living and washout plasma data are scarce. Some metabolites have been overlooked, notably phenyl-lactic, phenyl-hydracrylic and phenyl-propanoic acids, especially those from amino acids plus glycine, hydroxy-glycine and glutamine conjugates. Phenolic acids and conjugates from multiple sources exhibit biological activities, some of which are likely relevant in vivo and link to biomarkers of health. Further targeted studies are justified.
The present study attempted for the first time to investigate the metabolic fate of (poly)phenolic compounds provided by a hull-less and purple grain barley genotype biofortified in anthocyanins. Balb/c mice...
A bergamot-based beverage does not affect glucose, lipid and inflammatory biomarkers of cardiometabolic risk in healthy subjects.
Amino acids have been extensively studied in nutrition, mainly as key elements for maintaining optimal protein synthesis in the body as well as precursors of various nitrogen-containing compounds. However, it is now known that amino acid catabolism is an important element for the metabolic control of different biological processes, although it is still a developing field to have a deeper understanding of its biological implications. The mechanisms involved in the regulation of amino acid catabolism now include the contribution of the gut microbiota to amino acid oxidation and metabolite generation in the intestine, the molecular mechanisms of transcriptional control, and the participation of specific miRNAs involved in the regulation of amino acid degrading enzymes. In addition, molecules derived from amino acid catabolism play a role in metabolism as they are used in the epigenetic regulation of many genes. Thus, this review aims to examine the mechanisms of amino acid catabolism and to support the idea that this process is associated with the immune response, abnormalities during obesity, in particular insulin resistance, and the regulation of thermogenesis.
It is well established that p-Hydroxycinnamic acids (HCAs), including ferulic, caffeic, sinapic, and p-coumaric acids, possess a characteristic phenylpropanoid C6-C3 backbone and account for about one-third of the phenolic compounds in our diet. HCAs are typically associated with various plant cell wall components, including mono-, di-, and polysaccharides, sterols, polyamines, glycoproteins, and lignins. Interestingly, enzymes produced by intestinal microbes liberate HCAs from these associations. HCAs are completely absorbed in their free form upon ingestion and undergo specific reactions upon absorption in the small intestine or liver. The gut epithelium, composed of intestinal epithelial cells (IECs), acts as a physical barrier against harmful bacteria and a site for regulated interactions between bacteria and the gut lumen. Thus, maintaining the integrity of the epithelial barrier is essential for establishing a physiochemical environment conducive to homeostasis. This review summarizes the protective effects of HCAs on the intestinal barrier, achieved through four mechanisms: preserving tight junction proteins (TJPs), modulating pro-inflammatory cytokines, exerting antioxidant activity, and regulating the intestinal microbiota.
Background:
Dietary polyphenols, including flavan-3-ols (F3O), are associated with better health outcomes. The relationship of plasma phenyl-γ-valerolactones (PVLs), the products of colonic bacterial metabolism of F3O, with dietary intakes is unclear.
Objective:
To investigate whether plasma PVLs are associated with self-reported intakes of total F3O and procyanidins+(epi)catechins.
Design:
We measured 9 PVLs by uHPLC-MS-MS in plasma from adults (>60y) in the TUDA study (2008-2012; n=5,186) and a follow-up subset (2014-2018) with corresponding dietary data (n=557). Dietary (poly)phenols collected by FFQ were analyzed using Phenol-Explorer.
Results:
Mean (95% CI) intakes were estimated as 2283 (2213, 2352) mg/d for total (poly)phenols, 674 (648, 701) for total F3O, and 152 (146, 158) for procyanidins+(epi)catechins. Two PVL metabolites were detected in plasma from the majority of participants, 5-(hydroxyphenyl)-γ-VL-sulfate (PVL1) and 5-(4'-hydroxyphenyl)-γ-VL-3'-glucuronide (PVL2). The 7 other PVLs were detectable only in 1-32% of samples. Self-reported intakes (mg/d) of F3O (r=0.113, P=0.017) and procyanidin+(epi)catechin (r=0.122, P=0.010) showed statistically significant correlations with the sum of PVL1 and PVL 2 (PVL1+2). With increasing intake quartiles (Q1-Q4), mean (95% CI) PVL1+2 increased; from 28.3 (20.8, 35.9) nmol/L in Q1 to 45.2 (37.2, 53.2) nmol/L in Q4; P=0.025, for dietary F3O, and from 27.4 (19.1, 35.8) nmol/L in Q1 to 46.5 (38.2, 54.9) nmol/L in Q4; P=0.020, for procyanidins+(epi)catechins.
Conclusions:
Of 9 PVL metabolites investigated, two were detected in the majority of samples and were weakly associated with intakes of total F3O and procyanidins+(epi)catechins. Future controlled feeding studies are required to validate plasma PVLs as biomarkers of these dietary polyphenols.
Protocatechuic acid (PCA) is a natural component with multiple biological activities. However, the underlying mechanisms of the effects of PCA on anti-ulcerative colitis (UC) are unclear. A UC mouse model was established by allowing the mice to freely drink a dextran sulfate sodium solution. The mice were administered PCA intragastrically for 7 days. Histological pathology, intestinal flora, and ferroptosis regulators were determined in vivo. Additionally, ferroptotic Caco-2 cells were modeled to investigate the role of PCA in ferroptosis. Our results showed that PCA reduced the levels of the disease activity index, inflammatory factors, and histological damage in UC mice. We also found that the regulation of intestinal flora, especially Bacteroidetes, was one of the potential mechanisms underlying the protective effects of PCA anti-UC. Moreover, PCA downregulated the level of ferroptosis in the colon tissue, as evidenced by a reduced iron overload, decreased glutathione depletion, and a lower level of malondialdehyde production compared with the model group. Similar effects of PCA on ferroptosis were observed in Erastin-treated Caco-2 cells. The results obtained using reactive oxygen species assays and the changes in mitochondrial structure observed via scanning electron microscopy also support these results. Our findings suggested that PCA protected against UC by regulating intestinal flora and ferroptosis.
Background: Although widely used, there is limited understanding on the suitability of different dietary assessment tools to estimate (poly)phenol intake. This study aims to compare the agreement between a food...
Flavan-3-ols, including the flavan-3-ol monomer (-)-epicatechin, are dietary bioactives known to mediate beneficial cardiovascular effects in humans. Recent studies showed that flavan-3-ols could interact with methylxanthines, evidenced by an increase in flavan-3-ol bioavailability with a concomitant increase in flavan-3-ol intake-mediated vascular effects. This study aimed at elucidating flavan-3-ol-methylxanthine interactions in humans in vivo by evaluating the specific contributions of theobromine and caffeine on flavan-3-ol bioavailability. In ileostomists, the effect of methylxanthines on the efflux of flavan-3-ol metabolites in the small intestine was assessed, a parameter important to an understanding of the pharmacokinetics of flavan-3-ols in humans. In a randomized, controlled, triple cross-over study in volunteers with a functional colon (n = 10), co-ingestion of flavan-3-ols and cocoa methylxanthines, mainly represented by theobromine, increased peak circulatory levels (Cmax) of flavan-3-ols metabolites (+21 ± 8%; p < 0.05). Conversely, caffeine did not mediate a statistically significant effect on flavan-3-ol bioavailability (Cmax = +10 ± 8%, p = n.s.). In a subsequent randomized, controlled, double cross-over study in ileostomists (n = 10), cocoa methylxanthines did not affect circulatory levels of flavan-3-ol metabolites, suggesting potential differences in flavan-3-ol bioavailability compared to volunteers with a functional colon. The main metabolite in ileal fluid was (-)-epicatechin-3'-sulfate, however, no differences in flavan-3-ol metabolites in ileal fluid were observed after flavan-3-ol intake with and without cocoa methylxanthines. Taken together, these results demonstrate a differential effect of caffeine and theobromine in modulating flavan-3-ol bioavailability when these bioactives are co-ingested. These findings should be considered when comparing the effects mediated by the intake of flavan-3-ol-containing foods and beverages and the amount and type of methylxanthines present in the ingested matrixes. Ultimately, these insights will be of value to further optimize current dietary recommendations for flavan-3-ol intake. CLINICAL TRIAL REGISTRATION NUMBER: This work was registered at clinicaltrials.gov as NCT03526107 (study part 1, volunteers with functional colon) and NCT03765606 (study part 2, volunteers with an ileostomy).
c department of Food science and Health, instituto andaluz de investigacion y Formacion agraria Pesquera alimentaria y de la Produccion ecologica, sevilla, spain ABSTRACT ω-Phenyl-alkenoic acids are abundant in coffee, fruits, and vegetables. Along with ω-phenyl-alkanoic acids, they are produced from numerous dietary (poly)phenols and aromatic amino acids in vivo. This review addresses how phenyl-ring substitution and flux modulates their gut microbiota and endogenous β-oxidation. 3′,5′-Dihydroxy-derivatives (from alkyl-resorcinols, flavanols, proanthocyanidins), and 4′-hydroxy-phenolic acids (from tyrosine, p-coumaric acid, naringenin) are β-oxidation substrates yielding benzoic acids. in contrast, 3′,4′,5′-tri-substituted-derivatives, 3′,4′-dihydroxy-derivatives and 3′-methoxy-4′-hydroxy-derivatives (from coffee, tea, cereals, many fruits and vegetables) are poor β-oxidation substrates with metabolism diverted via gut microbiota dehydroxylation, phenylvalerolactone formation and phase-2 conjugation, possibly a strategy to conserve limited pools of coenzyme A. 4′-Methoxy-derivatives (citrus fruits) or 3′,4′-dimethoxy-derivatives (coffee) are susceptible to hepatic "reverse" hydrogenation suggesting incompatibility with enoyl-CoA-hydratase. Gut microbiota-produced 3′-hydroxy-4′-methoxy-derivatives (citrus fruits) and 3′-hydroxy-derivatives (numerous (poly)phenols) are excreted as the phenyl-hydracrylic acid β-oxidation intermediate suggesting incompatibility with hydroxy-acyl-CoA dehydrogenase, albeit with considerable inter-individual variation. Further investigation is required to explain inter-individual variation, factors determining the amino acid to which C 6-C 3 and C 6-C 1 metabolites are conjugated, the precise role(s) of l-carnitine, whether glycine might be limiting, and whether phenolic acid-modulation of β-oxidation explains how phenolic acids affect key metabolic conditions, such as fatty liver, carbohydrate metabolism and insulin resistance.
Evidence from in vitro, animal, and human studies links citrus fruit consumption with several health‐promoting effects. However, many in vitro studies disregard bioavailability data, a key factor determining responses in humans. Citrus (poly)phenol metabolism and bioavailability follow specific pathways that vary widely among individuals and are affected by several intrinsic (age, sex, gut microbiota, metabolic state, genetic polymorphisms) and extrinsic (food matrix, co‐consumed food, (poly)phenol solubility, dose, food processing, lifestyle) factors. The gut microbiota is crucial to both absorption of citrus (poly)phenols and the production of catabolites, and absorption of both takes place mostly in the colon. Citrus (poly)phenol absorption can reach up to 100% in some individuals when the sum of the gut microbiota products are taken into account. This review emphasizes the importance of understanding citrus (poly)phenol absorption, metabolism, and bioavailability using evidence primarily derived from human studies in designing in vitro, animal, and further human clinical studies.
A large number of epidemiological studies have shown that consumption of fruits, vegetables, and beverages rich in (poly)phenols promote numerous health benefits from cardiovascular to neurological diseases. Evidence on (poly)phenols has been applied mainly to flavonoids, yet the role of phenolic acids has been largely overlooked. Such phenolics present in food combine with those resulting from gut microbiota catabolism of flavonoids and chlorogenic acids and those produced by endogenous pathways, resulting in large concentrations of low molecular weight phenolic metabolites in human circulation. Independently of the origin, in human intervention studies using diets rich in (poly)phenols, a total of 137 low molecular weight phenolic metabolites have been detected and quantified in human circulation with largely unknown biological function. In this review, we will pinpoint two main aspects of the low molecular weight phenolic metabolites: (i) the microbiota responsible for their generation, and (ii) the analysis (quali‐ and quantitative) in human circulation and their respective pharmacokinetics. In doing so, we aim to drive scientific advances regarding the ubiquitous roles of low molecular weight phenolic metabolites using physiologically relevant concentrations and under (patho)physiologically relevant conditions in humans.
Scope:
Epidemiological evidence suggests that a reduced risk of colorectal cancer (CRC) is correlated with high consumption of fruits and vegetables, which are major sources of fiber and phytochemicals, such as flavan-3-ols. However, it remains unknown how these phytochemicals and their specific gut-related metabolites may alter cancer cell behavior.
Methods and results:
A focused screening using native (poly)phenols and gut microbial metabolites (GMMs) on 3D HCT116 spheroids was carried out using a high-throughput imaging approach. Dose-responses, IC50, and long-term exposure were calculated for the most promising native (poly)phenols and GMMs. As a result, this research shows that (poly)phenol catabolites may play a key role in preventing cancer propagation. Indeed, μM concentration levels of (4R)-5-(3',4'-dihydroxyphenyl)-γ-valerolactone significantly decreased spheroid size at early stages of spheroid aggregation and gene expression of matrix metalloproteinases.
Conclusion:
A chronic exposure to (4R)-5-(3',4'-dihydroxyphenyl)-γ-valerolactone may lead to a reduced CRC risk. Daily intake of monomeric, oligomeric, and polymeric flavan-3-ols might increase the colonic concentrations of this metabolite, and, in turn, this compound may act locally interacting with intestinal epithelial cells, precancerous and cancer cells. This article is protected by copyright. All rights reserved.
Background
Although older adults commonly take multivitamin-multimineral (MVM) supplements to promote health, evidence on use of daily MVMs on invasive cancer is limited.
Objectives
The study objective was to determine if a daily MVM decreases total invasive cancer among older adults.
Methods
We performed a randomized, double-blind, placebo-controlled, two-by-two factorial trial of a daily MVM and cocoa extract for prevention of cancer and cardiovascular disease (CVD) among 21,442 U.S. adults (12,666 women aged ≥ 65 years and 8776 men aged ≥ 60 years) free of major CVD and recently diagnosed cancer. The intervention phase was from June 2015 through December 2020. This article reports on the MVM intervention.
Intervention
Participants were randomly assigned to daily MVM or placebo. The primary outcome was total invasive cancer, excluding non-melanoma skin cancer. Secondary outcomes included major site-specific cancers, total CVD, all-cause mortality, and total cancer risk among those with a baseline history of cancer.
Results
During a median follow-up of 3.6 years, invasive cancer occurred in 518 participants in the MVM group and 535 participants in the placebo group (hazard ratio [HR]=0.97, 95% confidence interval [CI]: 0.86, 1.09; P = 0.57). We observed no significant effect of a daily MVM on breast cancer (HR = 1.06, 95% CI: 0.79, 1.42) or colorectal cancer (HR = 1.30, 95% CI: 0.80, 2.12). We observed a protective effect of a daily MVM on lung cancer (HR = 0.62, 95% CI: 0.42, 0.92). The composite CVD outcome occurred in 429 participants in the MVM group and 437 participants in the placebo group (HR = 0.98, 95% CI: 0.86, 1.12). MVM did not significantly affect all-cause mortality (HR = 0.93; 95% CI: 0.81,1.08). There were no safety concerns.
Conclusion
A daily MVM supplement, compared with placebo, did not significantly reduce the incidence of total cancer among older men and women. Future studies are needed to determine the effects of MVMs on other aging-related outcomes among older adults.
Urolithins, metabolites produced by the gut microbiota from the polyphenols ellagitannins and ellagic acid, were discovered by our research group in humans almost twenty years ago. Pioneering research suggested urolithins as pleiotropic bioactive contributors to explain the health benefits after consuming ellagitannin‐rich sources (pomegranates, walnuts, strawberries, etc.). Here, we comprehensively update the knowledge on urolithins, emphasizing the review of the literature published during the last five years. To date, thirteen urolithins and their corresponding conjugated metabolites (glucuronides, sulfates, etc.) have been described and, depending on the urolithin, detected in different human fluids and tissues (urine, blood, feces, breastmilk, prostate, colon, and breast tissues). There has been a substantial advance in the research on microorganisms involved in urolithin production, along with the compositional and functional characterization of the gut microbiota associated with urolithins metabolism that gives rise to the so‐called urolithin metabotypes (UM‐A, UM‐B and UM‐0), relevant in human health. The design of in vitro studies using physiologically relevant assay conditions (molecular forms and concentrations) is still a pending subject, making some reported urolithin activities questionable. In contrast, remarkable progress has been made in the research on the safety, bioactivity, and associated mechanisms of urolithin A, including the first human interventions. This article is protected by copyright. All rights reserved
Scope:
The study evaluated the influence of flavan-3-ol structure on the production of phenolic catabolites, principally phenyl-γ-valerolactones (PVLs) and phenylvaleric acids (PVAs).
Methods and results:
A set of 12 monomeric flavan-3-ols and proanthocyanidins (degree of polymerisation (DP) of 2-5), were fermented in vitro for 24 h using human faecal microbiota, and catabolism was analysed by UHPLC-ESI-MS/MS. Up to 32 catabolites strictly related to microbial catabolism of parent compounds were detected. (+)-Catechin and (-)-epicatechin had the highest molar mass recoveries, expressed as a percentage with respect to the incubated concentration (75 μmol L-1 ) of the parent compound, for total PVLs and PVAs, both at 5 h (about 20%) and 24 h (about 40%) of faecal incubation. Only A-type dimer and B-type procyanidins underwent the ring fission step, and no differences were found in total PVL and PVA production (≃ 1.5% and 6.0% at 5 h and 24 h faecal incubation, respectively) despite the different DPs.
Conclusion:
The flavan-3-ol structure strongly affected the colonic catabolism of the native compounds, influencing the profile of PVLs and PVAs produced in vitro. This study opens new perspectives to further elucidate the colonic fate of oligomeric flavan-3-ols and their availability in producing bioactive catabolites. This article is protected by copyright. All rights reserved.
Cocoa flavanols and procyanidins (CFs), natural dietary bioactives, have been studied extensively over the past two decades for their potential health benefits. Research on their safety and efficacy is critically dependent upon on the ability to reliably characeterize the research materials that are utilized, and with growing consumer availability of CF-based products, reliable methods for the detection of potential adulteration are of increasing importance. This research focused on the development of a high performance liquid chromatography-tandem mass spectrometry method (HPLC-MS²) using primary standards and ¹³C-labelled procyanidins as internal standards. The ability of MS² detection to discriminate A- and B-type procyanidins was demonstrated. Method performances were validated for degrees of polymerization up to four in seven model food matrices. Accuracy ranged from 90.9 to 125.4% and precision was < 10% at lower concentrations. Finally, the method was applied to cocoa-based samples and compared to the AOAC 2020.05 analytical protocol, supporting the use of NIST 8403 as reference material for HPLC-MS² analysis.
With the world's population aging, age-related memory decline is an impending cognitive epidemic. Assessing the impact of diet on cognitive aging, we conducted a controlled, randomized, parallel-arm dietary intervention with 211 healthy adults (50–75 years) investigating effects of either a placebo or 260, 510 and 770 mg/day of cocoa flavanols for 12-weeks followed by 8-weeks washout. The primary outcome was a newly-developed object-recognition task localized to the hippocampus’ dentate gyrus. Secondary outcomes included a hippocampal-dependent list-learning task and a prefrontal cortex-dependent list-sorting task. The alternative Healthy Eating Index and a biomarker of flavanol intake (gVLM) were measured. In an MRI substudy, hippocampal cerebral blood volume was mapped. Object-recognition and list-sorting performance did not correlate with baseline diet quality and did not improve after flavanol intake. However, the hippocampal-dependent list-learning performance was directly associated with baseline diet quality and improved after flavanol intake, particularly in participants in the bottom tertile of baseline diet quality. In the imaging substudy, a region-of-interest analysis was negative but a voxel-based-analysis suggested that dietary flavanols target the dentate gyrus. While replication is needed, these findings suggest that diet in general, and dietary flavanols in particular, may be associated with memory function of the aging hippocampus and normal cognitive decline.
Diets rich in fruit and vegetables are associated with a decreased incidence of colorectal cancer (CRC) due, in part, to the bioactive (poly)phenolic components and their microbiota-mediated metabolites. This study investigated how such compounds, derived from ingested raspberries in the gastrointestinal tract, may exert protective effects by reducing DNA damage. Ileal fluids collected pre- and post-consumption of 300 g of raspberries by ileostomists (n = 11) were subjected to 24 h ex vivo fermentation with fecal inoculum to simulate interaction with colonic microbiota. The impact of fermentation on (poly)phenolics in ileal fluid was determined and the bioactivity of ileal fluids pre- and post fermentation investigated. (Poly)phenolic compounds including sanguiin H-6, sanguiin H-10 and cyanidin-3-O-sophoroside decreased significantly during fermentation while, in contrast, microbial catabolites, including 3-(3′-hydroxyphenyl)propanoic acid, 3-hydroxybenzoic acid and benzoic acid increased significantly. The post-raspberry ileal fermentate from 9 of the 11 ileostomates significantly decreased DNA damage (∼30 %) in the CCD 841 CoN normal cell line using an oxidative challenge COMET assay. The raspberry ileal fermentates also modulated gene expression of the nuclear factor 2–antioxidant responsive element (Nrf2-ARE) pathway involved in oxidative stress cytoprotection, namely Nrf2, NAD(P)H dehydrogenase, quinone-1 and heme oxygenase-1. Four of the phenolic catabolites were assessed individually, each significantly reducing DNA damage from an oxidative challenge over a physiologically relevant 10-100 μM range. They also induced a differential pattern of expression of key genes in the Nrf2-ARE pathway in CCD 841 CoN cells. The study indicates that the colon-available raspberry (poly)phenols and their microbial-derived catabolites may play a role in protection against CRC in vivo.
Urinary polyphenol metabolites are potential biomarkers of dietary polyphenol intake. The current study aims to evaluate associations between total diet, vegetable and fruit polyphenol intakes with urinary polyphenol metabolite concentrations in a sample of adults prescribed a diet rich in vegetables and fruit. Thirty-four participants completed a 10-week pre-post study. Participants were asked to consume Australian recommended daily vegetable and fruit serves and attend measurement sessions at baseline and at weeks 2 and 10. Two 24-h diet recalls were collected at each time-point and polyphenol intakes were calculated using the Phenol-Explorer database. Spot urine samples, collected at each time-point, were analyzed for 15 polyphenol metabolites using liquid chromatography-mass spectroscopy. Spearman’s correlation analyzes assessed the strength of relationships between urinary and dietary polyphenols. Linear mixed models were used to investigate relationships between polyphenol excretion and intake. Total urinary polyphenols were significantly correlated with total polyphenol intakes at week 10 (rs = 0.47) and fruit polyphenols at week 2 (rs = 0.38). Hippuric acid was significantly correlated with vegetable polyphenols at baseline (rs = 0.39). Relationships were identified between individual polyphenol metabolites and vegetable and fruit polyphenols. Linear mixed model analyzes identified that for every 1 mg increase in polyphenol intakes, urinary polyphenol excretion increased by 16.3 nmol/g creatinine. Although the majority of relationships were not sufficiently strong or consistent at different time-points, promising relationships were observed between total urinary polyphenols and total polyphenol intakes, and hippuric acid and vegetable polyphenols.
Flavan-3-ols are a group of bioactive compounds that have been shown to improve vascular function in intervention studies. They are therefore of great interest for the development of dietary recommendation for the prevention of cardio-vascular diseases. However, there are currently no reliable data from observational studies, as the high variability in the flavan-3-ol content of food makes it difficult to estimate actual intake without nutritional biomarkers. In this study, we investigated cross-sectional associations between biomarker-estimated flavan-3-ol intake and blood pressure and other CVD risk markers, as well as longitudinal associations with CVD risk in 25,618 participants of the European Prospective Investigation into Cancer (EPIC) Norfolk cohort. High flavan-3-ol intake, achievable as part of an habitual diet, was associated with a significantly lower systolic blood pressure (− 1.9 (− 2.7; − 1.1) mmHg in men and − 2.5 (− 3.3; − 1.8) mmHg in women; lowest vs highest decile of biomarker), comparable to adherence to a Mediterranean Diet or moderate salt reduction. Subgroup analyses showed that hypertensive participants had stronger inverse association between flavan-3-ol biomarker and systolic blood pressure when compared to normotensive participants. Flavanol intake could therefore have a role in the maintenance of cardiovascular health on a population scale.
There is a lack of focus on the protective health effects of phy-tochemicals in dietary guidelines. Although a number of chemical libraries and databases contain dietary phytochemicals belonging to the plant metabolome, they are not entirely relevant to human health because many constituents are extensively metabolized within the body following ingestion. This is especially apparent for the highly abundant dietary (poly)phenols, for which the situation is compounded by confusion regarding their bioavailability and metabolism, partially because of the variety of nomenclatures and trivial names used to describe compounds arising from microbial catabolism in the gastrointestinal tract. This confusion, which is perpetuated in online chemical/metabolite databases, will hinder future discovery of bioactivities and affect the establishment of future dietary guidelines if steps are not taken to overcome these issues. In order to resolve this situation, a nomenclature system for phenolic catabolites and their human phase II metabolites is proposed in this article and the basis of its format outlined. Previous names used in the literature are cited along with the recommended nomenclature,
Tyrosine, phenylalanine and tryptophan are the three aromatic amino acids (AAA) involved in protein synthesis. These amino acids and their metabolism are linked to the synthesis of a variety of secondary metabolites, a subset of which are involved in numerous anabolic pathways responsible for the synthesis of pigment compounds, plant hormones and biological polymers, to name a few. In addition, these metabolites derived from the AAA pathways mediate the transmission of nervous signals, quench reactive oxygen species in the brain, and are involved in the vast palette of animal coloration among others pathways. The AAA and metabolites derived from them also have integral roles in the health of both plants and animals. This review delineates the de novo biosynthesis of the AAA by microbes and plants, and the branching out of AAA metabolism into major secondary metabolic pathways in plants such as the phenylpropanoid pathway. Organisms that do not possess the enzymatic machinery for the de novo synthesis of AAA must obtain these primary metabolites from their diet. Therefore, the metabolism of AAA by the host animal and the resident microflora are important for the health of all animals. In addition, the AAA metabolite-mediated host-pathogen interactions in general, as well as potential beneficial and harmful AAA-derived compounds produced by gut bacteria are discussed. Apart from the AAA biosynthetic pathways in plants and microbes such as the shikimate pathway and the tryptophan pathway, this review also deals with AAA catabolism in plants, AAA degradation via the monoamine and kynurenine pathways in animals, and AAA catabolism via the 3-aryllactate and kynurenine pathways in animal-associated microbes. Emphasis will be placed on structural and functional aspects of several key AAA-related enzymes, such as shikimate synthase, chorismate mutase, anthranilate synthase, tryptophan synthase, tyrosine aminotransferase, dopachrome tautomerase, radical dehydratase, and type III CoA-transferase. The past development and current potential for interventions including the development of herbicides and antibiotics that target key enzymes in AAA-related pathways, as well as AAA-linked secondary metabolism leading to antimicrobials are also discussed.
Age-related complications such as neurodegenerative disorders are increasing and remain cureless. The possibility of altering the progression or the development of these multifactorial diseases through diet is an emerging and attractive approach with increasing experimental support. We examined the potential of known bioavailable phenolic sulfates, arising from colonic metabolism of berries, to influence hallmarks of neurodegenerative processes. In silico predictions and in vitro transport studies across blood-brain barrier (BBB) endothelial cells, at circulating concentrations, provided evidence for differential transport, likely related to chemical structure. Moreover, endothelial metabolism of these phenolic sulfates produced a plethora of novel chemical entities with further potential bioactivies. Pre-conditioning with phenolic sulfates improved cellular responses to oxidative, excitotoxicity and inflammatory injuries and this attenuation of neuroinflammation was achieved via modulation of NF-κB pathway. Our results support the hypothesis that these small molecules, derived from dietary (poly)phenols may cross the BBB, reach brain cells, modulate microglia-mediated inflammation and exert neuroprotective effects, with potential for alleviation of neurodegenerative diseases.
Data obtained with in vitro fecal incubations and a feeding study indicate black tea theaflavin and its galloyl derivatives are not absorbed in detectable amounts in either the upper or the lower gastrointestinal tract. The theaflavin skeleton is comparatively resistant to degradation by colonic bacteria with a 67% recovery being obtained after a 24 h incubation which yielded 21 phenolic and aromatic catabolites. The theaflavin galloyl moiety was removed by the microbiota and the released gallic acid further transformed to 3-O-and 4-O-methyl gallic acids, pyrogallol-1-sulfate and pyrogallol-2-sulfate, which were excreted in urine in amounts equivalent to 94% of intake. The main urinary product potentially derived from breakdown of the theaflavin skeleton was 3-(4′-hydroxyphenyl)propionic acid. A number of the colonic catabolites originating from gallic acid and theaflavins have been reported to be bioactive in ex vivo and in vitro models with a variety of potential modes of action.
PurposeOrange juice (OJ) flavanones undergo limited absorption in the upper gastrointestinal tract and reach the colon where they are transformed by the microbiota prior to absorption. This study investigated the ability of two probiotic bacteria, Bifidobacterium longum R0175 and Lactobacillus rhamnosus subsp. Rhamnosus NCTC 10302 to catabolise OJ flavanones. Methods
The bacteria were incubated with hesperetin-7-O-rutinoside, naringenin-7-O-rutinoside, hesperetin and naringenin, and the culture medium and intracellular cell extracts were collected at intervals over a 48 h of incubation period. The flavanones and their phenolic acid catabolites were identified and quantified by HPLC–HR–MS. ResultsBoth probiotics were able to subject hesperetin to ring fission yielding 3-(3′-hydroxy-4′-methoxyphenyl)propionic acid which was subsequently demethylated producing 3-(3′,4′-dihydroxyphenyl)propionic acid and then via successive dehydroxylations converted to 3-(3′-hydroxyphenyl)propionic acid and 3-(phenyl)propionic acid. Incubation of both bacteria with naringenin resulted in its conversion to 3-(4′-hydroxyphenyl)propionic acid which underwent dehydroxylation yielding 3-(phenyl)propionic acid. In addition, only L. rhamnosus exhibited rhamnosidase and glucosidase activity and unlike B. longum, which was able to convert hesperetin-7-O-rutinoside and naringenin-7-O-rutinoside to their respective aglycones. The aglycones were then subjected to ring fission and further catabolised in a similar manner to that described above. The flavanones and their catabolites were found in the culture medium but not accumulated in the bacterial cells. Conclusions
These findings demonstrate the enzymatic potential of single strains of bifidobacterium and lactobacillus which may be involved in the colonic catabolism of OJ flavanones in vivo.
Diet is a major life style factor affecting human health, thus emphasizing the need for evidence-based dietary guidelines for primary disease prevention. While current recommendations promote intake of fruit and vegetables, we have limited understanding of plant-derived bioactive food constituents other than those representing the small number of essential nutrients and minerals. This limited understanding can be attributed to some extent to a lack of fundamental data describing the absorption, distribution, metabolism and excretion (ADME) of bioactive compounds. Consequently, we selected the flavanol (−)-epicatechin (EC) as an example of a widely studied bioactive food constituent and investigated the ADME of [2-14C](−)-epicatechin (300 μCi, 60 mg) in humans (n = 8). We demonstrated that 82 ± 5% of ingested EC was absorbed. We also established pharmacokinetic profiles and identified and quantified >20 different metabolites. The gut microbiome proved to be a key driver of EC metabolism. Furthermore, we noted striking species-dependent differences in the metabolism of EC, an insight with significant consequences for investigating the mechanisms of action underlying the beneficial effects of EC. These differences need to be considered when assessing the safety of EC intake in humans. We also identified a potential biomarker for the objective assessment of EC intake that could help to strengthen epidemiological investigations.
Urolithins are dibenzo[b,d]pyran-6-one derivatives that are produced by the human gut microbiota from ellagitannins and ellagic acid. These metabolites are much better absorbed than their precursors and have been suggested to be responsible for the health effects attributed to ellagitannins and ellagic acid that occur in food products as berries and nuts. In the present review, the role and potential of urolithins in human health are critically reviewed, and a perspective of the research approach needed to demonstrate these health effects is presented, based on the existing knowledge. The analytical methods available for urolithin analysis, their occurrence in different tissues and biological fluids and their metabolism by human gut microbiota are considered. In addition, the inter-individual variability observed for the production of urolithins (metabotypes), and its relationship with health status and dysbiosis are also reviewed. The potential mechanisms of action of urolithins are also critically discussed paying attention to the concentration and the type of metabolites used in the in vitro and in vivo assays and the physiological significance of the results obtained. The gut microbiota metabolism of ellagic acid to urolithins and that of daidzein to equol, their individual variations and the effects on health are also compared. This article is protected by copyright. All rights reserved.
Background: Flavonoids have been implicated in the prevention of cardiovascular disease; however, their mechanisms of action have yet to be elucidated, possibly because most previous in vitro studies have used supraphysiological concentrations of unmetabolized flavonoids, overlooking their more bioavailable phenolic metabolites.
Objective: We aimed to explore the effects of phenolic metabolites and their precursor flavonoids at physiologically achievable concentrations, in isolation and combination, on soluble vascular cellular adhesion molecule-1 (sVCAM-1).
Method: Fourteen phenolic acid metabolites and 6 flavonoids were screened at 1 μM for their relative effects on sVCAM-1 secretion by human umbilical vein endothelial cells stimulated with tumor necrosis factor alpha (TNF-α). The active metabolites were further studied for their response at different concentrations (0.01 μM–100 μM), structure-activity relationships, and effect on vascular cellular adhesion molecule (VCAM)-1 mRNA expression. In addition, the additive activity of the metabolites and flavonoids was investigated by screening 25 unique mixtures at cumulative equimolar concentrations of 1 μM.
Results: Of the 20 compounds screened at 1 μM, inhibition of sVCAM-1 secretion was elicited by 4 phenolic metabolites, of which protocatechuic acid (PCA) was the most active (−17.2%, P = 0.05). Investigations into their responses at different concentrations showed that PCA significantly reduced sVCAM-1 15.2–36.5% between 1 and 100 μM, protocatechuic acid-3-sulfate and isovanillic acid reduced sVCAM-1 levels 12.2–54.7% between 10 and 100 μM, and protocatechuic acid-4-sulfate and isovanillic acid-3-glucuronide reduced sVCAM-1 secretion 27.6% and 42.8%, respectively, only at 100 μM. PCA demonstrated the strongest protein response and was therefore explored for its effect on VCAM-1 mRNA, where 78.4% inhibition was observed only after treatment with 100 μM PCA. Mixtures of the metabolites showed no activity toward sVCAM-1, suggesting no additive activity at 1 μM.
Conclusions: The present findings suggest that metabolism of flavonoids increases their vascular efficacy, resulting in a diversity of structures of varying bioactivity in human endothelial cells.
After an acute intake of 300 g of mango purée by 10 subjects, 0 and 24 h urine and plasma samples were analyzed by high-performance liquid chromatography-high-resolution mass spectrometry. The method was first validated for 44 reference polyphenols in terms of linearity, specificity, limits of detection and quantification, intra-day and inter-day precision, recovery, and matrix effects in two biological matrices. After method validation, a total of 94 microbial-derived phenolic catabolites, including 15 cinnamic acids, 3 phenylhydracrylic acids, 14 phenylpropanoic acids, 12 phenylacetic acids, 28 benzoic acids, 2 mandelic acids, 15 hydroxybenzenes, and 5 hippuric acid derivatives, were identified or tentatively identified in urine and/or plasma. These results establish the value of the UHPLC-HRMS protocol and the use of authentic standards to obtain a detailed and accurate picture of mango polyphenol metabolites, together with their phase II conjugated metabolites, in human bioavailability studies.
Phenolic catabolites excreted by fasting subjects with a functioning colon and ileostomists on a low (poly)phenol diet have been investigated. Urine was collected over a 12 h fasting period after adherence to a low (poly)phenol diet for 36 h. UHPLC-HR-MS quantified 77 phenolics. Some were present in the urine of both groups in similar trace amounts and others were excreted in higher amounts by participants with a colon indicating the involvement of the microbiota. Most were present in sub- or low-µmol amounts, but hippuric acid dominated accounting on average for 60% of the total for both volunteer categories indicating significant production from sources other than non-nutrient dietary (poly)phenols. The potential origins of the phenolics associated with the low (poly)phenol diet, include endogenous catecholamines, surplus tyrosine and phenylalanine, and washout of catabolites derived from pre-study intakes of non-nutrient dietary (poly)phenols.
The impact of β-glucan-rich oat bran on the bioavailability of orange juice (OJ) flavanones was investigated. Volunteers consumed 500 mL of OJ with and without 22 g of oat bran containing 6 g of β-glucan (OB-6). Urine collected 12 h prior to and over a 0-24 h period post-supplementation was analysed by UHPLC-HRMS. Sixteen flavanone metabolites and thirty-nine colon-derived phenolic catabolites were identified and quantified. The major compounds were hesperetin-3'-glucuronide, along with hippuric acids and the C6-C3 phenolic acids 3-(3'-hydroxy-4'-methoxyphenyl)hydracrylic acid and 3-(4'-hydroxy-3'-methoxyphenyl)propanoic acid. A marked reduction in the 0-24 h excretion of flavanone metabolites from 29.7 μmol (9.3% recovery) to 9.3 μmol (2.9% recovery), occurred following consumption of OB-6 compared to OJ. This appeared not to be an effect of fiber on the rate of transport in the upper gut. After consumption of OJ there was a 163 ± 15 μmol excretion of colon-derived phenolic catabolites, equivalent to 43% of (poly)phenol intake and following OB-6 intake there was a further significant 30% increase. The β-oat bran in OB-6 contained 5.8 μmol of free and 52 μmol of bound phenolic derivatives compared to 371 μmol of OJ (poly)phenolics. The elevated excretion of phenolics after OB-6 consumption appears not to be due to bound phenolics in the bran, rather it is consequence, principally, of a bran-mediated increase in the quantities of flavanones passing from the upper to the lower bowel where they were subjected to microbiota-mediated catabolism. CLINICAL TRIAL REGISTRATION NUMBER: . This trial was registered at clinicaltrials.gov as NCT04867655.
A R T I C L E I N F O Keywords: Flavan-3-ols Colonic catabolic and phase II metabolism [2-14 C](−)-Epicatechin Procyanidins Theaflavins A B S T R A C T Understanding the fate of ingested polyphenols is crucial in elucidating the molecular mechanisms underlying the beneficial effects of a fruit and vegetable-based diet. This review focuses on the colon microbiota-mediated transformation of the flavan-3-ols and the structurally related procyanidins found in dietary plant foods and beverages, plus the flavan-3-ol-derived theaflavins of black tea, and the post-absorption phase II metabolism of the gut microbiota catabolites. Despite significant advances in the last decade major analytical challenges remain. Strategies to address them are presented.
Dysfunction of blood-brain barrier formed by endothelial cells of cerebral blood vessels, plays a key role in development of neurodegenerative disorders. Epicatechin exerts vasculo-protective effects through genomic modifications, however molecular mechanisms of action, particularly on brain endothelial cells, are largely unknow. This study aimed to use a multi-omic approach (transcriptomics of mRNA, miRNAs and lncRNAs, and proteomics), to provide novel in-depth insights into molecular mechanisms of how metabolites affect brain endothelial cells under lipid-stressed (as a model of BBB dysfunction) at physiological concentrations. We showed that metabolites can simultaneously modulate expression of protein-coding, non-coding genes and proteins. Integrative analysis revealed interactions between different types of RNAs and form functional groups of genes involved in regulation of processing like VEGF-related functions, cell signaling, cell adhesion and permeability. Molecular modeling of genomics data predicted that metabolites decrease endothelial cell permeability, increased by lipotoxic stress. Correlation analysis between genomic modifications observed and genomic signature of patients with vascular dementia and Alzheimer's diseases showed opposite gene expression changes. Taken together, this study describes for the first time a multi-omic mechanism of action by which (−)-epicatechin metabolites could preserve brain vascular endothelial cell integrity and reduce the risk of neurodegenerative diseases.
Significance
Dysfunction of the blood-brain barrier (BBB), characterized by dysfunction of endothelial cells of cerebral blood vessels, result in an increase in permeability and neuroinflammation which constitute a key factor in the development neurodegenerative disorders. Even though it is suggested that polyphenols can prevent or delay the development of these disorders, their impact on brain endothelial cells and underlying mechanisms of actions are unknow. This study aimed to use a multi-omic approach including analysis of expression of mRNA, microRNA, long non-coding RNAs, and proteins to provide novel global in-depth insights into molecular mechanisms of how (−)-epicatechin metabolites affect brain microvascular endothelial cells under lipid-stressed (as a model of BBB dysfunction) at physiological relevant conditions. The results provide basis of knowledge on the capacity of polyphenols to prevent brain endothelial dysfunction and consequently neurodegenerative disorders.
This review focusses on the LC–MS characterisation and quantification of dietary (poly)phenols and their metabolites. It draws attention to errors, omissions, and misunderstandings that appear frequently in published papers, and suggests strategies for their avoidance. Aspects covered include the use of authentic standards and surrogate reference materials, the importance of collecting and archiving Total Ion Current MS data, the limitations of using on‐line compilations of accurate mass MS data to assign unknown components when multiple isomers are possible, and the often understated magnitude of person‐to‐person variation that may significantly impact at population level any potential health benefit. This article is protected by copyright. All rights reserved
The practitioner's dilemma in metabolite assignment can be described as follows: For compound and metabolite identification, should we follow strict guidelines using authentic standards only, or should we accept uncertainties in structure assignment of compounds with the certainty of consequential errors. These uncertainties arise due to limitation of software and databases in combination with the complexity of the human body fluid samples. This article is protected by copyright. All rights reserved
Scope
Wild strawberries (Fragaria vesca) are richer in (poly)phenols than common commercial strawberry varieties, e.g., Fragaria × ananassa. (Poly)phenols and their microbiota-derived metabolites are hypothesised to exert bioactivity within the human gut mucosa. To address this, the effects of wild strawberries were investigated with respect to their bioactivity and microbiota-modulating capacity using both in vitro and ex vivo approaches.
Methods and results
Ileal fluids collected pre- (0h) and post-consumption (8h) of 225 g wild strawberries by ileostomates (n = 5) and also in vitro digested strawberry varieties (Fragaria vesca and Fragaria × ananassa Duchesne) supernatants were collected. Subsequent fermentation of these supernatants using an in vitro batch culture proximal colon model revealed significant treatment-specific changes in microbiome community structure in terms of alpha but not beta diversity at 24 h. Nutri-kinetic analysis revealed a significant increase in the concentration of gut microbiota catabolites, including 3-(4hydroxyphenyl)propionic acid, 3-(3-hydroxyphenyl)propanoic acid and benzoic acid. Furthermore, post-berry ileal fermentates (24 h) significantly (p<0.01) decreased DNA damage (% Tail DNA, COMET assay) in both HT29 cells (∼45%) and CCD 841 CoN cells (∼25%) compared to untreated controls.
Conclusions
Post berry consumption fermentates exhibited increased overall levels of (poly)phenolic metabolites which retained their bioactivity, reducing DNA damage in colonocytes.
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p-Hydroxybenzaldehyde (HD), the main active component of Nostoc commune, has been shown to attenuate dextran sulfate sodium (DSS)-induced experimental colitis. However, it is still unclear whether HD or its metabolites exert a protective role in ulcerative colitis (UC). In this study, we showed that p-hydroxybenzoic acid (HA) is the active metabolite of HD and demonstrated the effects of HA in alleviating colitis and its potential mechanism. Mice with colitis induced by DSS were orally administered or intraperitoneally injected with therapeutic drugs (including HD, HA, mesalazine and PHTPP) for 10 days. Caco-2 cells were treated with TNF-α (10 ng/ml) and therapeutic drugs (including HA, mesalazine and PHTPP) for 24 h. The results showed that oral supplementation, but not intraperitoneal injection, of HD (40 mg/kg) significantly ameliorated DSS-induced colitis. Oral administration of HA (10-40 mg/kg) dose-dependently attenuated various colitis phenotypes in DSS-treated mice. Treatment with HA also decreased the expression of proinflammatory cytokines and increased the expression of tight junction proteins in mice and Caco-2 cells. However, combined treatment of HA with PHTPP, a highly selective antagonist of the estrogen receptor β (ERβ), did not show anti-colitis effects. Collectively, our results demonstrate that both HD and HA are active substances capable of inhibiting inflammatory responses and improving intestinal mucosal damage via the activation of ERβ signaling in DSS-induced colitis.
Artichokes are a rich source of (poly)phenols, mainly caffeoylquinic acids, but little is known about their bioavailability from this source. This study investigated the absorption, metabolism and excretion of (poly)phenols after sous-vide artichoke consumption (5776 µmol of (poly)phenols) by healthy volunteers. Seventy-six (poly)phenol metabolites were identified by UHPLC-MS/MS using authentic standards, including acyl-quinic acids plus C6–C3, C6–C1, C6–C2, C6–C2–N, C6–C0 metabolites, and their phase-II conjugates. The major metabolites were 3ʹ-methoxy-4ʹ-hydroxycinnamic acid, 3ʹ-methoxycinnamic acid-4ʹ-sulfate, and 4ʹ-hydroxycinnamic acid-3ʹ-sulfate, which appeared early in plasma (Tmax < 4 h); plus 3-(3ʹ-methoxy-4ʹ-hydroxyphenyl)propanoic acid, 3-(4ʹ-methoxyphenyl)propanoic acid-3ʹ-glucuronide, 3-(3ʹ-hydroxyphenyl) propanoic acid and hippuric acids, which appeared later (Tmax >6 h). The 24 h urinary recovery averaged 8.9% (molar basis) of the (poly)phenols consumed. Hepatic beta-oxidation of 3ʹ,4ʹ-dihydroxycinnamic acid and methylated conjugates occurred, but was limited (<0.04%). 3ʹ-Methylation exceeded 4ʹ-methylation and interindividual variability was high, especially for gut microbial metabolites (up to 168-fold).
A single oral dose of tyrosine (100 or 150 mg/kg) caused significant increases in urinary levels of norepinephrine (NE), epinephrine (E), dopamine (DA), 3-methoxy-4-hydroxyphenylglycol (MHPG), vanilmandelic acid (VMA), and homovanillic acid (HVA) during the first 2 hr after its ingestion; water administration failed to produce such changes. The temporal patterns of these increases paralleled those of previously described increases in plasma tyrosine. Since urinary catecholamines derive from peripheral sources, while the catecholamine metabolites in urine may reflect both CNS and peripheral catecholamine turnover, these findings indicate that tyrosine administration may accelerate catecholamine synthesis in and release from cells throughout the human body. Tyrosine may thus constitute a useful agent for treating central or peripheral disorders associated with insufficient release of catecholamines.
The hydrolyzable tannins ellagitannins (ETs) and ellagic acid (EA) are polyphenols present in food sources such as pomegranates, berries, and walnuts. However, they are poorly absorbed on consumption, but the gut microbiota metabolizes them. In recent decades, an extensive literature has attributed a wide range of beneficial effects to these natural compounds based on their biological activities, including antioxidant, chemopreventive, antiinflammatory, cardioprotective, etc. However, in the last decade, knowledge of their bioavailability and metabolism has prompted research into the physiological role of their in vivo metabolites generated by gut microbiota action, the urolithins, which have received recognition across the world as possible candidates for health benefits. This chapter summarizes the latest developments and knowledge on the occurrence, dietary intake, bioavailability and metabolism, and biological effects of ellagitannins and ellagic acid supplementation, paying particular attention to the activity of their gut microbiota‐derived metabolites – urolithins.
Covering: 1958 to June 2018 Phenyl-γ-valerolactones (PVLs) and their related phenylvaleric acids (PVAs) are the main metabolites of flavan-3-ols, the major class of flavonoids in the human diet. Despite their presumed importance, these gut microbiota-derived compounds have, to date, in terms of biological activity, been considered subordinate to their parent dietary compounds, the flavan-3-ol monomers and proanthocyanidins. In this review, the role and prospects of PVLs and PVAs as key metabolites in the understanding of the health features of flavan-3-ols have been critically assessed. Among the topics covered, are proposals for a standardised nomenclature for PVLs and PVAs. The formation, bioavailability and pharmacokinetics of PVLs and PVAs from different types of flavan-3-ols are discussed, taking into account in vitro and animal studies, as well as inter-individual differences and the existence of putative flavan-3-ol metabotypes. Synthetic strategies used for the preparation of PVLs are considered and the methodologies for their identification and quantification assessed. Metabolomic approaches unravelling the role of PVLs and PVAs as biomarkers of intake are also described. Finally, the biological activity of these microbial catabolites in different experimental models is summarised. Knowledge gaps and future research are considered in this key area of dietary (poly)phenol research.
Bioavailability or pharmacokinetics of isoavones is based on data from absorption, metabolism, distribution, and excretion (ADME) studies conducted both in humans and animals. Following the consumption of pure compounds, isoavonerich extracts, or foods/beverages containing high levels of isoavones, the parent compounds and their metabolites can be detected in plasma and urine of human volunteers. Numerous studies attest to the fact that following ingestion, soy iso-avones attain maximal plasma concentrations (Cmax) within 4-8 h (Tmax) and are then eliminated from the body through the bile and kidneys with a terminal elimination half-life (t1/2) that is on average 8 h (Setchell et al. 2001, 2003a, b; Cassidy 2006). Available data suggest that they are more efciently absorbed than other subclasses of avonoids, withCmax levels of 2 µM and meanrelative urinary excretions of 42% for daidzein and 16% forgenistein, after a 50 mg isoavone intake (Manach et al. 2005).
This study developed, optimized and validated an ultra-high-performance liquid chromatography–high-resolution mass spectrometry (UHPLC-HRMS) method to identify and quantify metabolites and microbial-derived catabolites in urine, plasma and feces of rats following ingestion of 50 mg of a red wine proanthocyanidin-rich extract. The method was validated for specificity, linearity, limit of detection (LD) and quantification (LQ), intra-day and inter-day precision, recovery and matrix effects, which were determined for 34 compounds in the three biological matrices. After method validation, three parent flavan-3-ols, four 5-carbon side chain ring fission metabolites, and 27 phenolic acid and aromatic catabolites were quantified in plasma, urine and feces after red wine proanthocyanidin intake. These results establish the value of the UHPLC-HRMS protocol in obtaining a detailed picture of proanthocyanidin metabolites and their microbial-derived catabolites, along with their phase II metabolites, in biological fluids of rat, and potentially in human clinical studies designed to evaluate the bioavailability of dietary flavan-3-ols.
This paper reviews pioneering human studies, their limitations and recent investigations on the absorption, metabolism, distribution and excretion (aka bioavailability) of (-)-epicatechin. Progress has been made possible by improvements in mass spectrometric detection when coupled to high performance liquid chromatography and through the increasing availability of authentic reference compounds of in vivo metabolites of (-)-epicatechin. Studies have shown that [2-(14)C](-)-epicatechin is absorbed in the small intestine with the 12 structural-related (-)-epicatechin metabolites (SREMs), mainly in the form of (-)-epicatechin-3'-O-glucuronide, 3'-O-methyl-(-)-epicatechin-5-sulfate and (-)-epicatechin-3'-sulfate, attaining sub-μmol/L peak plasma concentrations (Cmax) ∼1 h after ingestion. SREMs were excreted in urine over a 24 h period in amounts corresponding to 20% of (-)-epicatechin intake. On reaching the colon the flavan-3-ol undergoes microbiota-mediated conversions yielding the 5C-ring fission metabolites (5C-RFMs) 5-(hydroxyphenyl)-γ-valerolactones and 5-(hydroxyphenyl)-γ-hydroxyvaleric acids which appear in plasma as phase II metabolites with a Cmax of 5.8 h after intake and are excreted in quantities equivalent to 42% of the ingested (-)-epicatechin. Other catabolites excreted in 0-24 h urine in amounts equivalent to 28% of intake included 3-(3'-hydroxyphenyl)hydracrylic acid, hippuric acid and 3'-hydroxyhippuric acid. Overall (-)-epicatechin is highly bioavailable with urinary excretion indicating that 95% is absorbed and passes through the circulatory systems as a diversity of phase II metabolites. Rats produce a very different profile of SREMs than that of humans. These findings demonstrate that ex vivo studies investigating the mechanisms underlying the protective effects of (-)-epicatechin on human health should make use of physiological concentrations human of SREMs and 5C-RFMs, and not the parent (-)-epicatechin, with model systems derived from human cells. In epidemiological studies 5-(4'-hydroxyphenyl)-γ-valerolactone-3'-sulfate and 5-(4'-hydroxyphenyl)-γ-valerolactone-3'-O-glucuronide, the principal 5C-RFMs in both plasma and urine, could serve as key biomarkers of (-)-epicatechin intake.
Polyphenols are beneficial for health, but are metabolised after consumption. We compared the vasorelaxant capacity of twenty-one physiologically relevant polyphenol metabolites in isolated mouse arteries. Hesperetin, urolithins and ferulic acid-4-O-sulfate...
(Poly)phenols are a large group of compounds, found in food, beverages, dietary supplements and herbal medicines. Owing to their biological activities, absorption and metabolism of the most abundant compounds in humans are well understood. Both the chemical structure of the phenolic moiety and any attached chemical groups define whether the polyphenol is absorbed in the small intestine, or reaches the colon and is subject to extensive catabolism by colonic microbiota. Untransformed substrates may be absorbed, appearing in plasma primarily as methylated, sulfated and glucuronidated derivatives, with in some cases the unchanged substrate. Many of the catabolites are well absorbed from the colon and appear in the plasma either similarly conjugated, or as glycine conjugates, or in some cases unchanged.
Background & aims:
Urolithins are microbial metabolites produced after consumption of ellagitannin-containing foods such as pomegranates and walnuts. Parallel to isoflavone-metabolizing phenotypes, ellagitannin-metabolizing phenotypes (urolithin metabotypes A, B and 0; UM-A, UM-B and UM-0, respectively) can vary among individuals depending on their body mass index (BMI), but correlations between urolithin metabotypes (UMs) and cardiometabolic risk (CMR) factors are unexplored. We investigated the association between UMs and CMR factors in individuals with different BMI and health status.
Methods:
UM was identified using UPLC-ESI-qToF-MS in individuals consuming pomegranate or nuts. The associations between basal CMR factors and the urine urolithin metabolomic signature were explored in 20 healthy normoweight individuals consuming walnuts (30 g/d), 49 healthy overweight-obese individuals ingesting pomegranate extract (450 mg/d) and 25 metabolic syndrome (MetS) patients consuming nuts (15 g-walnuts, 7.5 g-hazelnuts and 7.5 g-almonds/d).
Results:
Correlations between CMR factors and urolithins were found in overweight-obese individuals. Urolithin-A (mostly present in UM-A) was positively correlated with apolipoprotein A-I (P ≤ 0.05) and intermediate-HDL-cholesterol (P ≤ 0.05) while urolithin-B and isourolithin-A (characteristic from UM-B) were positively correlated with total-cholesterol, LDL-cholesterol (P ≤ 0.001), apolipoprotein B (P ≤ 0.01), VLDL-cholesterol, IDL-cholesterol, oxidized-LDL and apolipoprotein B:apolipoprotein A-I ratio (P ≤ 0.05). In MetS patients, urolithin-A only correlated inversely with glucose (P ≤ 0.05). Statin-treated MetS patients with UM-A showed a lipid profile similar to that of healthy normoweight individuals while a poor response to lipid-lowering therapy was observed in MB patients.
Conclusions:
UMs are potential CMR biomarkers. Overweight-obese individuals with UM-B are at increased risk of cardiometabolic disease, whereas urolithin-A production could protect against CMR factors. Further research is warranted to explore these associations in larger cohorts and whether the effect of lipid-lowering drugs or ellagitannin-consumption on CMR biomarkers depends on individuals' UM.
Clinical trial registry numbers and websites:
NCT01916239 (https://clinicaltrials.gov/ct2/show/NCT01916239) and ISRCTN36468613 (http://www.isrctn.com/ISRCTN36468613).
Scope:
Consumption of products rich in flavan-3-ols, such as tea and cocoa, has been associated with decreased obesity, partially dependent on their capacity to enhance energy expenditure. Despite these phenolics having been reported to increase the thermogenic program in brown and white adipose tissue, flavan-3-ols are vastly metabolised in vivo to phenyl-γ-valerolactones. Therefore, we hypothesize that phenyl-γ-valerolactones may directly stimulate the differentiation and the activation of brown adipocytes.
Methods and results:
Immortalized brown pre-adipocytes were differentiated in presence of (R)-5-(3',4'-dihydroxyphenyl)-γ-valerolactone (VL1), (R)-5-(3'-hydroxyphenyl)-γ-valerolactone-4'-O-sulphate (VL2), (R)-5-phenyl-γ-valerolactone-3',4'-di-O-sulphate (VL3), at concentrations of 2 or 10μM, whereas fully differentiated brown adipocyte were treated acutely (6-24h). None of the treatments regulated the expression levels of the uncouple protein 1, nor of the main transcription factors involved in brown adipogenesis. Similarly, mitochondrial content was unchanged after treatments. Moreover these compounds did not display peroxisome proliferator-activated receptor γ-agonist activity, as evaluated by luciferase assay, and did not enhance norepinephrine-stimulated lipolysis in mature adipocytes. However, both VL1 and VL2 prevented oxidative stress caused by H2 O2 .
Conclusion:
Phenyl-γ-valerolactones and their sulphated forms do not influence brown adipocyte development or function at physiological or supraphysiological doses in vitro, but they are active protecting brown adipocytes from increased reactive oxygen species production. This article is protected by copyright. All rights reserved.
Orange juice is a rich source of (poly)phenols, in particular the flavanones hesperetin-7-O-rutinoside and naringenin-7-O-rutinoside. Following the acute consumption of 500 mL of orange juice containing 398 µmol of (poly)phenols by 12 volunteers, 0-24 h plasma and urine samples were analysed by targeted high-performance liquid chromatography-high resolution-mass spectrometry in order to identify flavanone metabolites and phenolic acid and aromatic catabolites. A total of 19 flavanone metabolites which comprised di-O-glucuronide, O-glucuronide, O-glucuronyl-sulfate, and sulfate derivatives of hesperetin, naringenin and eriodictyol, and 65 microbial-derived phenolic catabolites, such as phenylpropanoid, phenylpropionic, phenylacetic, benzoic and hydroxycarboxylic acids and benzenetriol and benzoylglycine derivatives, including free phenolics and phase II sulfate, glucuronide and methyl metabolites, were identified or partially identified in plasma and/or urine samples. The data obtained provide a detailed evaluation of the fate of orange juice (poly)phenols as they pass through the gastrointestinal tract, and are absorbed into circulatory systems prior to renal excretion. Potential pathways for these conversions are proposed.
Epidemiological studies show that consumption of plant foods is associated with lowered risk of chronic diseases such as cancer and cardiovascular disease (WHO 2003). Dietary recommendations are based partly on this epidemiological evidence. Plant foods consist of cereal grains, vegetables, legumes, fruits, and berries. These plant foods contain (poly)phenolic compounds, which have been discussed in previous chapters of this book.