{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T02:01:14Z","timestamp":1778810474169,"version":"3.51.4"},"reference-count":21,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,1,24]],"date-time":"2026-01-24T00:00:00Z","timestamp":1769212800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Natural Science Foundation of Zhejiang province of China","award":["LZ26C170002"],"award-info":[{"award-number":["LZ26C170002"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32472918"],"award-info":[{"award-number":["32472918"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2023YFD1301104"],"award-info":[{"award-number":["2023YFD1301104"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Zhejiang Shuren University Foundation","award":["2024R009"],"award-info":[{"award-number":["2024R009"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Objectives: Iron deficiency impairs intestinal mucosal structure and function, yet its impact on intestinal stem cells (ISCs) remains unclear. This study was therefore designed to examine how iron deficiency affects the proliferation and differentiation of ISCs. Methods: Iron-deficient mouse and enteroid models were established. Expression of key cell markers was analyzed using Western blot, qPCR, and immunofluorescence. Results: Iron deficiency led to structural impairment of the intestinal mucosa, characterized by decreased small intestinal villus height. In iron-deficient mice, expression of ChrA (enteroendocrine cell marker), Lyz (Paneth cell marker), and Muc2 (goblet cell marker) was significantly downregulated across duodenum, jejunum and ileum, whereas Vil1 (enterocyte marker) expression increased. Moreover, both Lgr5 (an ISC marker) expression and the number of Ki67-positive proliferating cells were significantly reduced, along with a decrease in Ki67 transcriptional levels under iron-deficient conditions. Similarly, deferoxamine (DFO)-treated enteroids showed fewer Lgr5-positive ISCs, downregulation of Lgr5, Lyz and Muc2, and upregulation of Vil1. RNA-seq further confirmed that iron deficiency skews ISC differentiation toward absorptive lineage. This shift was associated with modulation of the Notch signaling pathway: upregulation of the ligand Dll1, receptors Notch2 and Notch3, and the protease ADAM10, alongside downregulation of the negative regulator Atoh1. These findings indicate that Notch pathway activation promotes enterocyte differentiation under iron deprivation. Conclusions: Iron deficiency suppressed the proliferation of ISCs and induced their differentiation toward enterocytes, which is associated with the modulation of the Notch signaling pathway, providing a mechanistic insights for impaired intestinal repair and the potential for nutrient-targeted therapies.<\/jats:p>","DOI":"10.3390\/nu18030392","type":"journal-article","created":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T11:14:07Z","timestamp":1769426047000},"page":"392","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Iron Deficiency Inhibits the Proliferation of Intestinal Stem Cells and Induces Their Differentiation to Enterocytes"],"prefix":"10.3390","volume":"18","author":[{"given":"Yecheng","family":"Xu","sequence":"first","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Zhao","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shouchuan","family":"Jiang","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Han","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yi","family":"Zheng","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xi","family":"Qiao","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xin","family":"Wen","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanyuan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yunqin","family":"Li","sequence":"additional","affiliation":[{"name":"Analysis Center of Agrobiology and Environmental Science, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingxia","family":"Kong","sequence":"additional","affiliation":[{"name":"Shulan International Medical College, Zhejiang Shuren University, Hangzhou 310015, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9603-8366","authenticated-orcid":false,"given":"Huahua","family":"Du","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Nutrition and Breeding for High-Quality Animal Products, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bao, H., Wang, Y., Xiong, H., Xia, Y., Cui, Z., and Liu, L. (2024). Mechanism of iron ion homeostasis in intestinal immunity and gut microbiota remodeling. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25020727"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e455","DOI":"10.1016\/S2352-3026(22)00096-5","article-title":"The case for classical haematology: The impact of a name and the future of a field","volume":"9","author":"Loren","year":"2022","journal-title":"Lancet Haematol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Montoro-Huguet, M.A., Belloc, B., and Dom\u00ednguez-Cajal, M. (2021). Small and large intestine (I): Malabsorption of nutrients. Nutrients, 13.","DOI":"10.3390\/nu13041254"},{"key":"ref_4","first-page":"103323","article-title":"Breaking the cycle: Psychological and social dimensions of pediatric functional gastrointestinal disorders","volume":"14","author":"Saeed","year":"2025","journal-title":"World J. Clin. Pediatr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.jcmgh.2022.12.004","article-title":"Glycoprotein (GP) 96 is essential for maintaining intestinal epithelial architecture by supporting its self-renewal capacity","volume":"15","author":"Schwarzfischer","year":"2023","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1126\/science.abb6848","article-title":"Tissue regeneration: Reserve or reverse?","volume":"371","author":"Shivdasani","year":"2021","journal-title":"Science"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3056","DOI":"10.1016\/j.cell.2024.05.004","article-title":"Isthmus progenitor cells contribute to homeostatic cellular turnover and support regeneration following intestinal injury","volume":"187","author":"Malagola","year":"2024","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"101674","DOI":"10.1016\/j.redox.2020.101674","article-title":"Mitochondrial electron transport chain: Oxidative phosphorylation, oxidant production, and methods of measurement","volume":"37","author":"Braganza","year":"2020","journal-title":"Redox Biol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kropp, P.A., Wu, J., Reidy, M., Shrestha, S., Rhodehouse, K., Rogers, P., and Golden, A. (2021). Allele-specific mitochondrial stress induced by multiple mitochondrial dysfunctions syndrome 1 pathogenic mutations modeled in Caenorhabditis elegans. PLoS Genet., 17.","DOI":"10.1371\/journal.pgen.1009771"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Vogt, A.C.S., Arsiwala, T., Mohsen, M., Vogel, M., Manolova, V., and Bachmann, M.F. (2021). On iron metabolism and its regulation. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22094591"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"103160","DOI":"10.1016\/j.redox.2024.103160","article-title":"Differentiation of intestinal stem cells toward goblet cells under systemic iron overload stress are associated with inhibition of Notch signaling pathway and ferroptosis","volume":"72","author":"Zhao","year":"2024","journal-title":"Redox Biol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Yiannikourides, A., and Latunde-Dada, G.O. (2019). A short review of iron metabolism and pathophysiology of iron disorders. Medicines, 6.","DOI":"10.3390\/medicines6030085"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Rose, E.C., Odle, J., Blikslager, A.T., and Ziegler, A.L. (2021). Probiotics, prebiotics and epithelial tight junctions: A promising approach to modulate intestinal barrier function. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22136729"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1016\/j.devcel.2020.12.020","article-title":"Transit-amplifying cells coordinate changes in intestinal epithelial cell-type composition","volume":"56","author":"Sanman","year":"2021","journal-title":"Dev. Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"295","DOI":"10.4236\/ojgas.2021.1112029","article-title":"Expression and clinical significance of Ki67 in common gastrointestinal tumours","volume":"11","author":"Shi","year":"2021","journal-title":"Open J. Gastroenterol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Flores, S.R., Nelson, S., Woloshun, R.R., Wang, X., Ha, J.H., Lee, J.K., and Collins, J.F. (2021). Intestinal iron absorption is appropriately modulated to match physiological demand for iron in wild-type and iron-loaded Hamp (hepcidin) knockout rats during acute colitis. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0252998"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1016\/j.tjnut.2024.01.013","article-title":"Dietary iron is necessary to support proliferative regeneration after intestinal injury","volume":"154","author":"Huang","year":"2024","journal-title":"J. Nutr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"skab258","DOI":"10.1093\/jas\/skab258","article-title":"Effects of iron, vitamin A, and the interaction between the two nutrients on intestinal development and cell differentiation in piglets","volume":"99","author":"Zhou","year":"2021","journal-title":"J. Anim. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Qin, Y.C., Jin, C.L., Hu, T.C., Zhou, J.Y., Wang, X.F., Wang, X.Q., and Yan, H.C. (2024). Early weaning inhibits intestinal stem cell expansion to disrupt the intestinal integrity of duroc piglets via regulating the Keap1\/Nrf2 signaling. Antioxidants, 13.","DOI":"10.3390\/antiox13101188"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1038\/s41575-022-00675-x","article-title":"The role of goblet cells and mucus in intestinal homeostasis","volume":"19","author":"Gustafsson","year":"2022","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e00689-23","DOI":"10.1128\/jvi.00689-23","article-title":"Porcine deltacoronavirus infection disrupts the intestinal mucosal barrier and inhibits intestinal stem cell differentiation to goblet cells via the notch signaling pathway","volume":"97","author":"Zhang","year":"2023","journal-title":"J. Virol."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/18\/3\/392\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T13:09:54Z","timestamp":1769778594000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/18\/3\/392"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,24]]},"references-count":21,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["nu18030392"],"URL":"https:\/\/doi.org\/10.3390\/nu18030392","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,24]]}}}