Gut and blood microbiomics for studying the effect of a polyphenol-rich dietary pattern on intestinal permeability in the elderly

JPI HDHL “Intestinal Microbiomics” (IM2015)
Gut and blood microbiomics for studying the effect of a polyphenol-rich dietary pattern on intestinal permeability in the elderly
Patrizia Riso
University of Milan (UMIL)


Partner Organization Partner Country
Institute of Food Research (now Quadram Institute Bioscience - QIB)United Kingdom
University of Barcelona (UB)Spain

1. Overall project description

1.1 Summary

An increased intestinal permeability (IP), also known as leaky gut, could be responsible for activation of the immune system and inflammation, a common mechanism in many chronic age-related diseases. 

The MaPLE project tested the hypothesis that an increased intake of polyphenol-rich foods can reduce IP and the quantity of inflammogenic bacterial factors in the bloodstream, promoting a protective metabolic phenotype. A dietary protocol was developed to deliver three portions per day of specific polyphenol-rich foods/beverages (berries and derived products, blood oranges and juice, pomegranate juice, Renetta apple and purée, green tea and dark chocolate products) to older participants living in a controlled setting (i.e. nursing home and residencies).

The polyphenol-enriched diet caused reductions in IP (reduced serum zonulin) and the effect was greatest in participants with higher baseline IP and metabolic dysregulation. Moreover, a reduction of diastolic blood pressure was found.Food metabolome profiles were characterized by an increase of flavanols (present in cocoa), the flavanones hesperetin and naringenin (present in oranges) and specific products of microbial metabolism, such as coumaric acids and urolithins (derived from blueberries and pomegranate). After polyphenol-rich diet intervention, the reductions in IP (serum zonulin) were associated with increases in theobromine and two methylxanthine metabolites.  We were unable to detect significant changes in the overall faecal metabolites. However, it is noteworthy that, following the polyphenol-rich dietary pattern, we found a significant increase of microbial-diversity suggesting an improvement in the gut microbial ecosystem, a reduction of specific pro-inflammatory bacteria and an associated increase of protective microbial derived metabolites.

Mechanisms were investigated using cultured intestinal cell and animal models. Using very old mice, it was shown that supplementation with a mixture of polyphenols that were similar to those incorporated into the older human participant diets caused a significant reduction in IP. No decreases in IP were observed in the younger mice, suggesting that the polyphenol-mediated effects are restricted to the very old animals. The polyphenol-supplemented diet did not change the gut microbiota structure or faecal metabolite profiles and the gut immune response to a flagellin challenge. Using cultured intestinal cells, it was shown that specific flavonoids caused rapid and sustained increases in barrier function. In addition, specific polyphenols caused significant changes in the secretion of inflammatory cytokines in response to flagellin, a bacteria-derived inflammatory challenge. 

The MaPLE project will generate new data for the exploitation in terms of future dietary guidelines that can be easily applied in different contexts such as nursing homes or for the support in the management of different clinical conditions associated with increased IP.

1.2 Highlights

Aging is a societal challenge that need to be faced through multi target strategies including the important contribution of an adequate diet to healthy aging. Experimental evidence suggests that an alteration in human intestinal barrier properties can occur during aging. In this regard, the Maple project has produced several important findings and possibility of exploitation in different contexts as it is reported below.

- Intestinal permeability appeared to be increased in aged people and should be considered as a potential target for screening (e.g. of increased risk of inflammation, metabolic disorders and associated clinical conditions)

- Serum zonulin, a protein involved in the regulation of intercellular junctions, was correlated with circulating microbial DNA suggesting these as potential future evaluation as candidate biomarkers for age-related clinical conditions

- A polyphenol-rich diet had a positive impact on IP in a group of older subjects, and these findings were supported by similar findings in an animal model of the ageing gut

- We have generated evidence to support a plausible (but as yet unproven) mechanism of action, i.e. that the beneficial effects of the PR diet on IP were the result of decreases in a group of pro-inflammatory and IP-associated bacteria, and these changes were also associated with changes in important bacterial metabolites (SCFAs) associated with a healthy gut.

- In vitro, specific polyphenols were shown to (i) cause rapid and sustained increases in intestinal barrier function (flavonoid-induced increases in TEER) and (ii) reduce the expression of inflammatory cytokines such as IL-8 in flagellin-challenged CaCo2 cells (quercetin and genistein, but not epigallocatechin gallate).

- An increased polyphenol intake caused also a reduction in diastolic blood pressure and the effect was stronger in women.

- We generated data that boost the study of polyphenol metabolites derived from bacterial activity, in order to reflect the microbiota's footprint on nutritional biomarkers.

-The inhibition of the intestinal enzyme PARP1 by specific methylxanthine metabolites and microbial derived polyphenol metabolites could explain, at least in part, and at molecular scale, the positive effects of the MaPLE diet on the intestinal permeability.

-The development of an in-house polyphenol database will support estimations of the intakes of different classes of and individual polyphenols, and their food sources, in future cohort and RCT studies. It will also underpin efforts to develop new targeted approaches for dietary recommendations in older adults.

-Our study demonstrated that older subjects are available to include food products in their diet if this does not change dramatically their dietary habits. The results will impact on future dietary guidelines, the development of new food products and translational research including efforts to increase awareness of the general public

4. Impact

4.1 List of publications

AuthorsTitleYear, Issue, PPPartners NumberDoiPdf
Del Bo' C (UMIL), Bernardi S (UMIL), Marino M (UMIL), Porrini M (UMIL), Tucci M (UMIL), Guglielmetti S (UMIL), Cherubini A (INRCA linked to UMIL), Carrieri B (INRCA linked to UMIL), Kirkup B (QIB), Kroon PA (QIB), Zamora-Ros R (UB), Hidalgo Liberona N (UB), Andres-Lacueva C (UB), Riso P (UMIL)Systematic review on polyphenol intake and health outcomes: is there sufficient evidence to define a health-promoting polyphenol-rich dietary pattern?2019, 11(6), 135510.3390/nu11061355Download
Bernardi S (UMIL), Del Bo' C (UMIL), Marino M (UMIL), Gargari G (UMIL), Cherubini A (INRCA linked to UMIL), Andrés-Lacueva C (UB), Hidalgo-Liberona N (UB), Peron G (UB), González-Dominguez R (UB), Kroon P (QIB), Kirkup B (QIB), Porrini M (UMIL), Guglielmetti S (UMIL), Riso P (UMIL). Polyphenols and Intestinal Permeability: Rationale and Future Perspectives. 2020, 68(7), 1816-182910.1021/acs.jafc.9b02283Download
Peron G (UB), Hidalgo-Liberona N (UB), González-Domínguez R (UB), Garcia-Aloy M (UB), Guglielmetti S (UMIL), Bernardi S (UMIL), Kirkup B (QIB), Kroon PA (QIB), Cherubini A (INRCA linked to UMIL), Riso P (UMIL), Andrés-Lacueva C (UB). Exploring the Molecular Pathways Behind the Effects of Nutrients and Dietary Polyphenols on Gut Microbiota and Intestinal Permeability: A Perspective on the Potential of Metabolomics and Future Clinical Applications.2020, 68(7), 1780-178910.1021/acs.jafc.9b01687
Guglielmetti S (UMIL), Bernardi S (UMIL), Del Bo' C (UMIL), Cherubini A (INRCA linked to UMIL), Porrini M (UMIL), Gargari G (UMIL), Hidalgo-Liberona N (UB), Gonzalez-Dominguez R (UB), Peron G (UB), Zamora-Ros R (UB), Winterbone MS (QIB), Kirkup B (QIB), Kroon PA (QIB), Andres-Lacueva C (UB), Riso P(UMIL)Effect of a polyphenol-rich dietary pattern on intestinal permeability and gut and blood microbiomics in older subjects: study protocol of the MaPLE randomised controlled trial.2020, 20(1), 7710.1186/s12877-020-1472-9Download
Gonzalez-Dominguez R (UB), Urpi-Sarda M (UB), Jauregui O (UB), Needs PW (QIB), Kroon PA (QIB), Andres-Lacueva C (UB)Quantitative Dietary Fingerprinting (QDF)-A Novel Tool for Comprehensive Dietary Assessment Based on Urinary Nutrimetabolomics. 2019, 68, 1851-186110.1021/acs.jafc.8b07023
Gargari G (UMIL), Taverniti V (UMIL), Del Bo’C (UMIL), Bernardi S (UMIL), Andres-Lacueva C (UB), González-Domínguez R (UB), Kroon PA (QIB), Winterbone MS (QIB), Cherubini A (INRCA linked to UMIL), Riso P (UMIL), Guglielmetti S (UMIL).Bacterial DNAemia is associated with serum zonulin levels in older subjects. 202010.1101/2020.04.10.035519Download
Bernardi S (UMIL), Del Bo’ C (UMIL), Guglielmetti S (UMIL), A Cherubini (INRCA linked to UMIL), Kroon PA (QIB), Kirkup B (QIB), Hidalgo Liberona N (UB), Peron G (UB), González-Domíinguez R (UB), Andrés-Lacueva C (UB), Riso P (UB).Role of a Polyphenol-Rich Dietary Pattern in the Modulation of Intestinal Permeability in Older Subjects: The MaPLE Study.2019, 11(1), 810.3390/proceedings2019011008
Castellano-Escuder, P (UB); González-Domínguez, R (UB); Wishart, DS (UB); Andrés-Lacueva, C (UB), Sánchez-Pla, A (UB).FOBI: An ontology to represent food intake data and associate it with metabolomic data.2020
Martini D (UMIL), Marino M (UMIL), Angelino D, Del Bo' C (UMIL), Del Rio D, Riso P (UMIL), Porrini M (UMIL). Role of berries in vascular function: a systematic review of human intervention studies. 2020, 78(3), 189-20610.1093/nutrit/nuz053Download
Marino M (UMIL), Del Bo' C (UMIL), Tucci M (UMIL), Klimis-Zacas D, Riso P (UMIL), Porrini M (UMIL).Modulation of Adhesion Process, E-Selectin and VEGF Production by Anthocyanins and Their Metabolites in an in vitro Model of Atherosclerosis. 2020, 12(3)10.3390/nu12030655Download
Saha S (QIB), Kroon PA (QIB).A simple and rapid LC-MS/MS method for quantification of total daidzein, genistein and equol in urine. 2020, Article No. 235939710.1155/2020/2359397Download
Madrid-Gambin F (UB), Garcia-Aloy M (UB), Vázquez-Fresno R (UB), Vegas-Lozano E (UB), Sánchez-Pla A, Misawa K, Hase T, Shimotoyodome A, Andres-Lacueva C (UB). Metabolic signature of a functional high-catechin tea after acute and sustained consumption in healthy volunteers through 1H NMR-based metabolomics analysis of urine.2019, 67(11), 3118-312410.1021/acs.jafc.8b04198
Dalla Via A (UMIL), Gargari G (UMIL), Taverniti V (UMIL), Rondini G, Velardi I, Gambaro V, Visconti GL, De Vitis V, Gardana C (UMIL), Ragg E, Pinto A, Riso P (UMIL), Guglielmetti S (UMIL). , Ragg E, Pinto A, Riso P (UMIL), Guglielmetti S (UMIL). Urinary TMAO Levels Are Associated with the Taxonomic Composition of the Gut Microbiota and with the Choline TMA-Lyase Gene (cutC) Harbored by Enterobacteriaceae. 2019 12(1) 10.3390/nu12010062Download
Cattaneo C, Riso P (UMIL), Laureati M, Gargari G (UMIL), Pagliarini E. Exploring Associations between Interindividual Differences in Taste Perception, Oral Microbiota Composition, and Reported Food Intake. 2019, 11(5) 10.3390/nu11051167Download
Del Bo' C (UMIL), Marino M (UMIL), Martini D (UMIL), Tucci M (UMIL), Ciappellano S (UMIL), Riso P (UMIL), Porrini M (UMIL). Overview of Human Intervention Studies Evaluating the Impact of the Mediterranean Diet on Markers of DNA Damage.2019, 11(2)10.3390/nu11020391Download
Del Bo' C (UMIL), Marino M (UMIL), Riso P (UMIL), Møller P, Porrini M (UMIL). Anthocyanins and metabolites resolve TNF-α-mediated production of E-selectin and adhesion of monocytes to endothelial cells. 2019, 300, 49-55. 10.1016/j.cbi.2019.01.002Download
Casas-Agustench P (UB), Cherubini A (INRCA linked to UMIL), Andrés-Lacueva C (UB). Lipids and physical function in older adults. 2017 1, 16-25.10.1097/MCO.0000000000000333
Garcia-Aloy (UB) M, Hulshof PJM, Estruel-Amades S, Osté, MCJ, Lankinen M, Geleijnse JM, de Goede J, Ulaszewska M, Mattivi F, Bakker SJL, Schwab U, Andres-Lacueva C (UB). Biomarkers of food intake for nuts and vegetable oils: an extensive literature search. 2019, 14 10.1186/s12263-019-0628-8

4.2 Presentation of the project

Target groupAuthorsMeans of communicationHyperlinkPdf

4.3 List of submitted patents and other outputs

Patent licencePartners involvedYearInternational eu or national patentCommentPdf

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This project has received funding from the European Union’s
H2020 Research and Innovation Programme under grant agreement n.696300

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