{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T03:46:08Z","timestamp":1791431168387,"version":"4.3.3"},"reference-count":50,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T00:00:00Z","timestamp":1758240000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Modern Agroindustry Technology Research System of China","award":["CARS-34"],"award-info":[{"award-number":["CARS-34"]}]},{"name":"Modern Agroindustry Technology Research System of China","award":["22IRTSTHN022"],"award-info":[{"award-number":["22IRTSTHN022"]}]},{"name":"Science and Technology Innovation Team of Henan Province High Quality Forage and Animal Health","award":["CARS-34"],"award-info":[{"award-number":["CARS-34"]}]},{"name":"Science and Technology Innovation Team of Henan Province High Quality Forage and Animal Health","award":["22IRTSTHN022"],"award-info":[{"award-number":["22IRTSTHN022"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Background\/Objectives: Ulcerative colitis (UC) pathogenesis involves gut barrier dysfunction, dysregulated immune responses, and gut microbiota imbalance. Alfalfa polysaccharide (APS), a bioactive compound with immunomodulatory potential, remains underexplored in intestinal inflammation. While APS exhibits anti-inflammatory properties in vitro, its in vivo efficacy, mechanisms, and ability to restore gut microbiota and barrier integrity in UC are unclear. This study aims to investigate the treatment effect of APS on dextran sulfate sodium (DSS)-induced colitis in mice and confirm its prebiotic potential. Methods: A mouse model of ulcerative colitis was induced by DSS. RNA sequencing, Western blotting, the terminal deoxynucleotidyl transferase dUTP nick end labeling technique, and an immuno-histochemical technique were used to study the mechanism of action by which APS at different dosages relieves DSS-induced colitis. Results: The findings show that APS alleviated the symptoms of colitis in mice given DSS, improved the gut morphology, heightened goblet cells production, increased the levels of IL-10 and IL-22, decreased the levels of TNF-\u03b1, IL-1\u03b2, and IL-6, and prevented the activation of the TLR4\/MyD88\/NF-\u03baB pathways. Additionally, they maintained the integrity of the intestine by enhancing the expression of the mucins MUC2 and MUC5AC and by increasing the amounts of ZO-1, Occludin, and Claudin-1 proteins. Moreover, APS supported the growth of probiotic bacteria, including unclassified_f_lachnospiraceae, Parabacteroides, Alistipes, and Mucispirillum, and in particular, Parabacteroides distasonis, which is strongly associated with decreased pro-inflammatory cytokine through the inhibition of the TLR4-MyD88-NF\u03baB pathways. Conclusions: APS can be used as a new type of prebiotic to improve UC by regulating intestinal flora and enhancing intestinal barrier function against the TLR4-MyD88-NF\u03baB pathway.<\/jats:p>","DOI":"10.3390\/nu17183001","type":"journal-article","created":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T12:33:32Z","timestamp":1758285212000},"page":"3001","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Alfalfa Polysaccharide Alleviates Colitis by Regulating Intestinal Microbiota and the Intestinal Barrier Against the TLR4\/MyD88\/NF-\u03baB Pathway"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9308-2292","authenticated-orcid":false,"given":"Shaokai","family":"La","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad","family":"Abaidullah","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yalei","family":"Cui","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"},{"name":"Henan Key Laboratory of Innovation and Utilization of Grassland Resources, Zhengzhou 450046, China"},{"name":"Henan Forage Engineering Technology Research Center, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boshuai","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"},{"name":"Henan Key Laboratory of Innovation and Utilization of Grassland Resources, Zhengzhou 450046, China"},{"name":"Henan Forage Engineering Technology Research Center, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinghua","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"},{"name":"Henan Key Laboratory of Innovation and Utilization of Grassland Resources, Zhengzhou 450046, China"},{"name":"Henan Forage Engineering Technology Research Center, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.jnutbio.2017.12.002","article-title":"Dietary antioxidant micronutrients alter mucosal inflammatory risk in a murine model of genetic and microbial susceptibility","volume":"54","author":"Pierre","year":"2018","journal-title":"J. Nutr. Biochem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"17004","DOI":"10.1038\/nmicrobiol.2017.4","article-title":"Dynamics of the human gut microbiome in inflammatory bowel disease","volume":"2","author":"Halfvarson","year":"2017","journal-title":"Nat. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11947","DOI":"10.1080\/21655979.2021.2009322","article-title":"Improvement of intestinal barrier function, gut microbiota, and metabolic endotoxemia in type 2 diabetes rats by curcumin","volume":"12","author":"Huang","year":"2021","journal-title":"Bioengineered"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1097\/MIB.0b013e31827fec6d","article-title":"Phylogenetic analysis of dysbiosis in ulcerative colitis during remission","volume":"19","author":"Shanahan","year":"2013","journal-title":"Inflamm. Bowel Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"112010","DOI":"10.1016\/j.foodres.2022.112010","article-title":"Targeting the gut microbiota to investigate the mechanism of Lactiplantibacillus plantarum 1201 in negating colitis aggravated by a high-salt diet","volume":"162","author":"Chen","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1080\/19490976.2016.1190073","article-title":"Composition and function of the pediatric colonic mucosal microbiome in untreated patients with ulcerative colitis","volume":"7","author":"Shah","year":"2016","journal-title":"Gut Microbes"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s12276-018-0126-x","article-title":"Mechanisms regulating intestinal barrier integrity and its pathological implications","volume":"50","author":"Chelakkot","year":"2018","journal-title":"Exp. Mol. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"67","DOI":"10.2147\/CEG.S181590","article-title":"Beneficial effects of Saccharomyces boulardii CNCM I-745 on clinical disorders associated with intestinal barrier disruption","volume":"12","author":"Terciolo","year":"2019","journal-title":"Clin. Exp. Gastroenter"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"13282","DOI":"10.1021\/acs.jafc.9b05744","article-title":"Orally Administered CLA Ameliorates DSS-Induced Colitis in Mice via Intestinal Barrier Improvement, Oxidative Stress Reduction, and Inflammatory Cytokine and Gut Microbiota Modulation","volume":"67","author":"Chen","year":"2019","journal-title":"J. Agric. Food Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1016\/j.ijbiomac.2021.03.083","article-title":"Tremella fuciformis polysaccharides ameliorated ulcerative colitis via inhibiting inflammation and enhancing intestinal epithelial barrier function","volume":"180","author":"Xiao","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"113280","DOI":"10.1016\/j.jep.2020.113280","article-title":"Combination of polysaccharides from Astragalus membranaceus and Codonopsis pilosula ameliorated mice colitis and underlying mechanisms","volume":"264","author":"Tang","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2290213","DOI":"10.1080\/07853890.2023.2290213","article-title":"Lycium barbarum polysaccharide alleviates DSS-induced chronic ulcerative colitis by restoring intestinal barrier function and modulating gut microbiota","volume":"55","author":"Li","year":"2023","journal-title":"Ann. Med."},{"key":"ref_13","first-page":"868","article-title":"The proliferative effects of alfalfa polysaccharides on the mouse immune cells","volume":"10","author":"Li","year":"2013","journal-title":"Life Sci. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1016\/j.ijbiomac.2018.12.227","article-title":"Polysaccharide from alfalfa activates RAW 264.7 macrophages through MAPK and NF-\u03baB signaling pathways","volume":"126","author":"Xie","year":"2019","journal-title":"Int. Biol. Macromol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.ijbiomac.2020.01.239","article-title":"Structural features, selenization modification, antioxidant and anti-tumor effects of polysaccharides from alfalfa roots","volume":"149","author":"Gao","year":"2020","journal-title":"Int. Biol. Macromol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"130334","DOI":"10.1016\/j.ijbiomac.2024.130334","article-title":"Microbiological and metabolic pathways analysing the mechanisms of alfalfa polysaccharide and sulfated alfalfa polysaccharide in alleviating obesity","volume":"263","author":"Li","year":"2024","journal-title":"Int. Biol. Macromol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1016\/j.ijbiomac.2019.04.144","article-title":"Immunomodulatory, antioxidant and intestinal morphology-regulating activities of alfalfa polysaccharides in mice","volume":"133","author":"Xie","year":"2019","journal-title":"Int. Biol. Macromol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1093\/carcin\/23.6.993","article-title":"Inhibition of chronic ulcerative colitis-associated colorectal adenocarcinoma development in a murine model by N-acetylcysteine","volume":"23","author":"Seril","year":"2002","journal-title":"Carcinogenesis"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1002\/ibd.20489","article-title":"5-aminosalicylic acid inhibits colitis-associated colorectal dysplasias in the mouse model of azoxymethane\/dextran sulfate sodium-induced colitis","volume":"14","author":"Clapper","year":"2008","journal-title":"Inflamm. Bowel Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1002\/ibd.21403","article-title":"Importance of disrupted intestinal barrier in inflammatory bowel diseases","volume":"17","author":"Salim","year":"2011","journal-title":"Inflamm. Bowel Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1016\/j.ijbiomac.2022.12.155","article-title":"Ameliorative effect of buckwheat polysaccharides on colitis via regulation of the gut microbiota","volume":"227","author":"Yang","year":"2023","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"124689","DOI":"10.1016\/j.ijbiomac.2023.124689","article-title":"Angelica sinensis aboveground part polysaccharide and its metabolite 5-MT ameliorate colitis via modulating gut microbiota and TLR4\/MyD88\/NF-kappaB pathway","volume":"242","author":"Zou","year":"2023","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.livsci.2019.01.027","article-title":"Effects of dietary supplementation of alfalfa polysaccharides on growth performance, small intestinal enzyme activities, morphology, and large intestinal selected microbiota of piglets","volume":"223","author":"Zhang","year":"2019","journal-title":"Livest. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.17219\/acem\/64883","article-title":"Intestinal epithelial barrier: The target for pathogenic Escherichia coli","volume":"26","author":"Pawlowska","year":"2017","journal-title":"Adv. Clin. Exp. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"105231","DOI":"10.1016\/j.jff.2022.105231","article-title":"Dendrobium huoshanense polysaccharides ameliorate ulcerative colitis by improving intestinal mucosal barrier and regulating gut microbiota","volume":"96","author":"Yu","year":"2022","journal-title":"J. Funct. Foods"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"112630","DOI":"10.1016\/j.foodres.2023.112630","article-title":"Alleviative effects of natural plant polysaccharides against DSS-induced ulcerative colitis via inhibiting inflammation and modulating gut microbiota","volume":"167","author":"Lu","year":"2023","journal-title":"Food Res. Int."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.bbrc.2020.04.103","article-title":"Heat shock transcription factor 2 inhibits intestinal epithelial cell apoptosis through the mitochondrial pathway in ulcerative colitis","volume":"527","author":"Wang","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10690","DOI":"10.1039\/D0FO02260H","article-title":"A Ganoderma atrum polysaccharide alleviated DSS-induced ulcerative colitis by protecting the apoptosis\/autophagy-regulated physical barrier and the DC-related immune barrier","volume":"11","author":"Zheng","year":"2020","journal-title":"Food Funct."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Li, Z., Wang, H., Wang, Z., and Geng, Y. (2022). Pine Pollen Polysaccharides\u2019 and Sulfated Polysaccharides\u2019 Effects on UC Mice through Modulation of Cell Tight Junctions and RIPK3-Dependent Necroptosis Pathways. Molecules, 27.","DOI":"10.3390\/molecules27227682"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1038\/s41467-021-21277-2","article-title":"Pancreatic glycoprotein 2 is a first line of defense for mucosal protection in intestinal inflammation","volume":"12","author":"Kurashima","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"114108","DOI":"10.1016\/j.jep.2021.114108","article-title":"Aloe vera mitigates dextran sulfate sodium-induced rat ulcerative colitis by potentiating colon mucus barrier","volume":"279","author":"Shi","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1002\/ptr.6866","article-title":"The role of medicinal products in the treatment of inflammatory bowel diseases (IBD) through inhibition of TLR4\/NF-kappaB pathway","volume":"35","author":"Dejban","year":"2021","journal-title":"Phytother. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2211","DOI":"10.1016\/j.fshw.2023.03.041","article-title":"Stachyose modulates gut microbiota and alleviates DSS-induced ulcerative colitis in mice","volume":"12","author":"Wang","year":"2023","journal-title":"Food Sci. Hum. Wellness"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/1471-2180-11-7","article-title":"High-throughput clone library analysis of the mucosa-associated microbiota reveals dysbiosis and differences between inflamed and non-inflamed regions of the intestine in inflammatory bowel disease","volume":"11","author":"Walker","year":"2011","journal-title":"BMC Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1053\/j.gastro.2011.04.011","article-title":"Twin Study Indicates Loss of Interaction Between Microbiota and Mucosa of Patients with Ulcerative Colitis","volume":"141","author":"Lepage","year":"2011","journal-title":"Gastroenterology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"104489","DOI":"10.1016\/j.phrs.2019.104489","article-title":"Phloretin ameliorates dextran sulfate sodium-induced ulcerative colitis in mice by regulating the gut microbiota","volume":"150","author":"Wu","year":"2019","journal-title":"Pharmacol. Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Abaidullah, M., La, S., Liu, M.Q., Liu, B.S., Cui, Y.L., Wang, Z.C., Sun, H., Ma, S., and Shi, Y.H. (2023). Polysaccharide from Roxb Mitigates Intestinal Mucosal Damage by Therapeutically Restoring the Interactions between Gut Microbiota and Innate Immune Functions. Nutrients, 15.","DOI":"10.3390\/nu15194102"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1038\/s41564-021-01050-3","article-title":"Multi-omics analyses of the ulcerative colitis gut microbiome link Bacteroides vulgatus proteases with disease severity","volume":"7","author":"Mills","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2112","DOI":"10.1136\/gutjnl-2023-330291","article-title":"Roseburia intestinalis generated butyrate boosts anti-PD-1 efficacy in colorectal cancer by activating cytotoxic CD8(+) T cells","volume":"72","author":"Kang","year":"2023","journal-title":"Gut"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1635","DOI":"10.1136\/gutjnl-2022-329386","article-title":"Parabacteroides distasonis: An emerging probiotic?","volume":"72","author":"Wu","year":"2023","journal-title":"Gut"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"118231","DOI":"10.1016\/j.carbpol.2021.118231","article-title":"Ganoderma lucidum polysaccharide modulates gut microbiota and immune cell function to inhibit inflammation and tumorigenesis in colon","volume":"267","author":"Guo","year":"2021","journal-title":"Carbohyd. Polym."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"118921","DOI":"10.1016\/j.carbpol.2021.118921","article-title":"Polysaccharide from edible alga Gloiopeltis furcata attenuates intestinal mucosal damage by therapeutically remodeling the interactions between gut microbiota and mucin O-glycans","volume":"278","author":"Pan","year":"2022","journal-title":"Carbohydr. Polym."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1136\/gutjnl-2012-302578","article-title":"Connecting dysbiosis, bile-acid dysmetabolism and gut inflammation in inflammatory bowel diseases","volume":"62","author":"Duboc","year":"2013","journal-title":"Gut"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"13780","DOI":"10.1073\/pnas.0706625104","article-title":"Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases","volume":"104","author":"Frank","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1016\/j.ijbiomac.2020.03.053","article-title":"Anti-inflammatory properties and gut microbiota modulation of an alkali-soluble polysaccharide from purple sweet potato in DSS-induced colitis mice","volume":"153","author":"Sun","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/j.jad.2021.06.006","article-title":"Ingestion of Faecalibaculum rodentium causes depression-like phenotypes in resilient Ephx2 knock-out mice: A role of brain-gut-microbiota axis via the subdiaphragmatic vagus nerve","volume":"292","author":"Wang","year":"2021","journal-title":"J. Affect. Disord."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e00102-16","DOI":"10.1128\/mSphere.00102-16","article-title":"Metabolic and Community Synergy of Oral Bacteria in Colorectal Cancer","volume":"1","author":"Flynn","year":"2016","journal-title":"Msphere"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"4322","DOI":"10.1038\/s41467-020-17956-1","article-title":"Integrated microbiota and metabolite profiles link Crohn\u2019s disease to sulfur metabolism","volume":"11","author":"Metwaly","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1002\/ijc.31559","article-title":"Parabacteroides distasonis attenuates toll-like receptor 4 signaling and Akt activation and blocks colon tumor formation in high-fat diet-fed azoxymethane-treated mice","volume":"143","author":"Koh","year":"2018","journal-title":"Int. J. Cancer"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1080\/14712598.2019.1666101","article-title":"Modern use of 5-aminosalicylic acid compounds for ulcerative colitis","volume":"20","author":"Roda","year":"2020","journal-title":"Expert Opin. Biol. Ther."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/18\/3001\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:49:07Z","timestamp":1760035747000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/18\/3001"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,19]]},"references-count":50,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["nu17183001"],"URL":"https:\/\/doi.org\/10.3390\/nu17183001","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,19]]}}}