Butyrate-Rich Diets Improve Redox Status And Fibrin Lysis In Behcet'S Syndrome

HomeCirculation ResearchVol. 128, No. 2Butyrate-Rich Diets Improve Redox Status and Fibrin Lysis in Behçet’s Syndrome Free AccessLetterPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyRedditDiggEmail Jump toFree AccessLetterPDF/EPUBButyrate-Rich Diets Improve Redox Status and Fibrin Lysis in Behçet’s Syndrome Giacomo Emmi, Alessandra Bettiol, Elena Niccolai, Matteo Ramazzotti, Amedeo Amedei, Giuditta Pagliai, Niccolò Taddei, Francesco Sofi, Claudia Fiorillo, Domenico Prisco and Matteo Becatti Giacomo EmmiGiacomo Emmi https://orcid.org/0000-0001-9575-8321 Department of Experimental and Clinical Medicine (G.E., A.B., E.N., A.A., G.P., F.S., D.P.), University of Firenze, Italy. Search for more papers by this author , Alessandra BettiolAlessandra Bettiol https://orcid.org/0000-0002-2292-1412 Department of Experimental and Clinical Medicine (G.E., A.B., E.N., A.A., G.P., F.S., D.P.), University of Firenze, Italy. Department of Neurosciences, Psychology, Drug Research and Child Health (NEUROFARBA) (A.B.), University of Firenze, Italy. Search for more papers by this author , Elena NiccolaiElena Niccolai https://orcid.org/0000-0002-9205-8079 Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, (M.R., N.T., C.F., M.B.), University of Firenze, Italy. Search for more papers by this author , Matteo RamazzottiMatteo Ramazzotti https://orcid.org/0000-0002-6862-9409 Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, (M.R., N.T., C.F., M.B.), University of Firenze, Italy. Search for more papers by this author , Amedeo AmedeiAmedeo Amedei https://orcid.org/0000-0002-6797-9343 Department of Experimental and Clinical Medicine (G.E., A.B., E.N., A.A., G.P., F.S., D.P.), University of Firenze, Italy. Search for more papers by this author , Giuditta PagliaiGiuditta Pagliai https://orcid.org/0000-0002-2177-2857 Department of Experimental and Clinical Medicine (G.E., A.B., E.N., A.A., G.P., F.S., D.P.), University of Firenze, Italy. Search for more papers by this author , Niccolò TaddeiNiccolò Taddei https://orcid.org/0000-0003-2513-1018 Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, (M.R., N.T., C.F., M.B.), University of Firenze, Italy. Search for more papers by this author , Francesco SofiFrancesco Sofi https://orcid.org/0000-0001-7113-7424 Department of Experimental and Clinical Medicine (G.E., A.B., E.N., A.A., G.P., F.S., D.P.), University of Firenze, Italy. Don Carlo Gnocchi Foundation, Onlus IRCCS, Firenze, Italy (F.S.). Search for more papers by this author , Claudia FiorilloClaudia Fiorillo Correspondence to: Claudia Fiorillo, PhD, Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, University of Firenze, Viale Morgagni 50, 50134 Firenze, Italy. Email E-mail Address: [email protected] https://orcid.org/0000-0003-1165-1581 Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, (M.R., N.T., C.F., M.B.), University of Firenze, Italy. Search for more papers by this author , Domenico PriscoDomenico Prisco https://orcid.org/0000-0002-7787-9610 Department of Experimental and Clinical Medicine (G.E., A.B., E.N., A.A., G.P., F.S., D.P.), University of Firenze, Italy. Search for more papers by this author and Matteo BecattiMatteo Becatti https://orcid.org/0000-0002-2593-5908 Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, (M.R., N.T., C.F., M.B.), University of Firenze, Italy. Search for more papers by this author Originally published17 Nov 2020https://doi.org/10.1161/CIRCRESAHA.120.317789Circulation Research. 2021;128:278–280is related toMeet the First AuthorsOther version(s) of this articleYou are viewing the most recent version of this article. Previous versions: November 17, 2020: Ahead of Print Meet the First Author, see p 154Behçet’s syndrome (BS) is a rare neutrophilic vasculitis characterized by arterial and venous events, uniquely treated by immunosuppressants rather than anticoagulants. We demonstrated in BS an impaired fibrinogen structure and fibrin susceptibility to plasmin-induced lysis,1 mainly because of an enhanced neutrophils reactive oxygen species (ROS) production, supporting the notion that BS is a model of inflammation-induced thrombosis.1In BS, we showed a peculiar gut microbiota (GM) fingerprint, mainly represented by a depletion of Roseburia and Subdoligranulum (Clostridium cluster), involved in short-chain fatty acids (SCFAs) production and a consequent reduction in fecal SCFAs, especially butyrate.2 Preclinical evidence suggests that GM-derived products, particularly SCFAs, might be involved in cardiovascular prevention.3This proof-of-concept randomized trial assessed the effects of 2 butyrate-enriched diets on blood redox status and fibrin degradation in BS, also considering GM alterations and clinical modifications.Seventeen omnivore White BS patients (aged 45.6±10.8 years, 9 males) without traditional cardiovascular risk factors or events, on stable corticosteroids and colchicine treatment, and without infections, antibiotics and/or pre-/probiotics use in the previous 3 months, were enrolled. The sample size was based on paired test for continuous outcomes, assuming a change for intracellular ROS of 1000±750 relative fluorescence units, with 80% power.After ethical approval and informed consent, 8 patients were randomized to habitual diet supplemented with oral butyrate (2.4 g/day), whereas 9 to a lacto-ovo-vegetarian diet containing insulin and resistant starch-rich foods (ie, cold potatoes, stale bread, and chicory) whose fermentation increases butyrate production.4Before and after dietary interventions (T0–T3), we evaluated blood global redox status by assaying intracellular leukocytes ROS production, plasma lipid peroxidation, and plasma total antioxidant capacity.1 On purified fibrinogen fractions, fibrin susceptibility to plasmin-induced lysis was evaluated after 6 hours of plasmin digestion.1 Values were compared at T3 versus T0 using the Wilcoxon signed-rank test for paired data.GM composition and SCFAs production were evaluated on fecal samples at T0 and T3 (in 6 and 7 patients, respectively), using next-generation sequencing of 16S rRNA and agilent gas chromatography-mass spectrometry, respectively.Clinical response was assessed based on disease activity (by the Behçet’s Disease Current Activity Form), corticosteroid use, and blood inflammatory parameters.Baseline demographic, clinical, and laboratory characteristics were comparable between the 2 groups. Both butyrate-enriched interventions alike showed a significant reduction in leukocyte ROS production (Figure [A] and [B]), and in plasma lipid peroxidation (Figure [C]) and an increase in plasma total antioxidant capacity (Figure [D]; for all analysis: P=0.0117 and P=0.0077 for the butyrate supplementation and the lacto-ovo-vegetarian group, respectively). Notably, a significant improvement in fibrin susceptibility to plasmin-induced lysis was observed in both groups (Figure [E] and [F]).Download figureDownload PowerPointFigure. Effect of a tailored butyrate-enriched dietary intervention on oxidative stress markers, fibrinogen clot degradation, gut microbiota composition, and clinical parameters in Behçet’s syndrome (BS). Intracellular lymphocyte/monocyte/neutrophil reactive oxygen species (ROS) production (A and B), plasma lipid peroxidation in terms of MDA (malondialdehyde) content (C), and plasma total antioxidant capacity (D), in patients with BS with oral butyrate supplementation or lacto-ovo-vegetarian diet, before (T0) and after the intervention (T3). Intracellular leukocyte ROS production was assessed by flow cytometry. Specifically, 100 µL of EDTA-anticoagulated blood samples were re-suspended in 2 mL of BD FACS lysing solution (Becton-Dickinson), gently mixed and incubated at room temperature in the dark for 10 min. After centrifugation, cells were washed twice in PBS and incubated with H2DCF-DA (2.5 µmol/L; Invitrogen) in RPMI medium without serum and phenol red for 15 min at 37 °C. After labeling, cells were washed and re-suspended in PBS and immediately analyzed using FACS Canto flow cytometer (Becton-Dickinson). E, Representative gel of plasmin-dependent fibrin degradation and relative residual fibrin beta-chain intensity (F) of fibrinogen purified from patients with BS, at T0 and T3. Fecal gut microbiota composition at phylum level before (T0) and after (T3) oral butyrate supplementation (G) or lacto-ovo-vegetarian diet (H). I, BS current activity form (BDCAF), daily dose of prednisolone equivalents (mg/d; J), C-reactive protein levels (mg/dL; K), and erythrocyte sedimentation rate (ESR; L) assessed before (T0) and after (T3) oral butyrate supplementation or lacto-ovo-vegetarian diet. P were calculated by Wilcoxon signed-rank test.Differently, no significant changes in GM composition were observed (Figure [G] and [H]), although more members of the Clostridium XIVa, Roboutsia, and Eggerthella genera were detected after butyrate supplementation. Also, no significant differences in SCFA production were found (data not shown).Both interventions led to a significant reduction in BS disease activity and corticosteroids use (Figure [I] and [J]), although without significant changes in blood inflammatory parameters (Figure [K] and [L]).In conclusion, this proof-of-concept trial shows for the first time that 2 butyrate-enriched diets modulate blood redox status and promote fibrin degradation, which is impaired by a neutrophil-dependent mechanism (via ROS) in BS.1 Although disease activity significantly improved, a 3-month butyrate-enriched diet did not affect GM composition and SCFAs production,5 suggesting that longer nutritional interventions are needed for scratching microbial resilience.A randomized crossover 3-arms trial is ongoing to elucidate the benefits of long-term butyrate-enriched interventions in BS. Pending these results, our preliminary data suggest the benefits of tailored dietary interventions for cardiovascular prevention.Nonstandard Abbreviations and AcronymsBSBehçet’s syndromeGMgut microbiotaROSreactive oxygen speciesSCFAshort-chain fatty acidSources of FundingThis study was supported by the Associazione Italiana Sindrome e Malattia di Behçet (SIMBA).Disclosures The data that support the findings of this study are available from the corresponding author upon reasonable request.FootnotesRegistration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT03274648, NCT03962335.For Sources of Funding and Disclosures, see page 279.Correspondence to: Claudia Fiorillo, PhD, Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, University of Firenze, Viale Morgagni 50, 50134 Firenze, Italy. Email claudia.[email protected]itReferences1. Becatti M, Emmi G, Silvestri E, Bruschi G, Ciucciarelli L, Squatrito D, Vaglio A, Taddei N, Abbate R, Emmi L, et al.. Neutrophil activation promotes fibrinogen oxidation and thrombus formation in behçet disease.Circulation. 2016; 133:302–311. doi: 10.1161/CIRCULATIONAHA.115.017738LinkGoogle Scholar2. Consolandi C, Turroni S, Emmi G, Severgnini M, Fiori J, Peano C, Biagi E, Grassi A, Rampelli S, Silvestri E, et al.. Behçet’s syndrome patients exhibit specific microbiome signature.Autoimmun Rev. 2015; 14:269–276. doi: 10.1016/j.autrev.2014.11.009CrossrefMedlineGoogle Scholar3. Brown JM, Hazen SL. Microbial modulation of cardiovascular disease.Nat Rev Microbiol. 2018; 16:171–181. doi: 10.1038/nrmicro.2017.149CrossrefMedlineGoogle Scholar4. Jung TH, Jeon WM, Han KS. In vitro effects of dietary inulin on human fecal microbiota and butyrate production.J Microbiol Biotechnol. 2015; 25:1555–1558. doi: 10.4014/jmb.1505.05078CrossrefMedlineGoogle Scholar5. Pagliai G, Russo E, Niccolai E, Dinu M, Di Pilato V, Magrini A, Bartolucci G, Baldi S, Menicatti M, Giusti B, et al.. Influence of a 3-month low-calorie Mediterranean diet compared to the vegetarian diet on human gut microbiota and SCFA: the CARDIVEG Study.Eur J Nutr. 2020; 59:2011–2024. doi: 10.1007/s00394-019-02050-0CrossrefMedlineGoogle Scholar Previous Back to top Next FiguresReferencesRelatedDetailsCited By Wang G, Li X, Li N, Wang X, He S, Li W, Fan W, Li R, Liu J and Hou S (2022) Icariin alleviates uveitis by targeting peroxiredoxin 3 to modulate retinal microglia M1/M2 phenotypic polarization, Redox Biology, 10.1016/j.redox.2022.102297, 52, (102297), Online publication date: 1-Jun-2022. Emmi G, Bagni G, Lastraioli E, Di Patti F, Bettiol A, Fiorillo C, Becatti M, Silvestri E, Urban M, Emmi L, Prisco D and Arcangeli A (2021) A unique circulating miRNA profile highlights thrombo-inflammation in Behçet’s syndrome, Annals of the Rheumatic Diseases, 10.1136/annrheumdis-2021-220859, 81:3, (386-397), Online publication date: 1-Mar-2022. Bettiol A, Becatti M, Silvestri E, Argento F, Fini E, Mannucci A, Galora S, Mattioli I, Urban M, Malandrino D, Palermo A, Taddei N, Emmi G, Prisco D and Fiorillo C (2021) Neutrophil-mediated mechanisms of damage and in-vitro protective effect of colchicine in non-vascular Behçet's syndrome , Clinical and Experimental Immunology, 10.1111/cei.13664, 206:3, (410-421), Online publication date: 2-Nov-2021. 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