{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T05:02:24Z","timestamp":1789448544807,"version":"build-2803163510"},"reference-count":39,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32502957"],"award-info":[{"award-number":["32502957"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32502957"],"award-info":[{"award-number":["32502957"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Jiangsu Provincial Academy of Environmental Science","award":["BK20251517"],"award-info":[{"award-number":["BK20251517"]}]},{"award":["BK20251517"],"award-info":[{"award-number":["BK20251517"]}],"id":[{"id":"https:\/\/ror.org\/03cvc1094","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Acute inflammatory challenge can impair intestinal barrier function and bile acid homeostasis in weaned piglets. Glutamate serves as an important energy substrate and functional amino acid for intestinal epithelial cells, yet the epitranscriptomic mechanisms underlying its potential protective effects remain poorly defined. This study investigated whether dietary glutamate supplementation is associated with m6A-dependent regulation of SLC10A2 and bile acid signaling in weaned piglets subjected to an acute LPS challenge, with supporting validation in IPEC-J2 cells. LPS challenge (100 \u03bcg\/kg body weight) impaired ileal morphological structure, activated the TLR4\/NF-\u03baB signaling pathway, and aggravated intestinal inflammatory responses in piglets. In contrast, glutamate supplementation was associated with restored ileal mucosal morphology, up-regulated expression of intestinal tight junction proteins, and suppressed TLR4\/NF-\u03baB pathway activation. Further metabolomic analysis indicated that LPS stimulation and glutamate intervention altered ileal bile acid metabolism: LPS decreased total bile acid (TBA) content in ileal tissue while increasing TBA accumulation in ileal chyme, whereas glutamate treatment partially reversed this pattern, by elevating ileal tissue TBA and reducing chyme TBA, suggesting a facilitatory effect of glutamate on intestinal bile acid reabsorption. Glutamate supplementation was also associated with increased ileal ASBT (SLC10A2) and FXR (NR1H4) expression. The SLC10A2 transcript exhibited detectable m6A modification; glutamate supplementation rescued the LPS-induced downregulation of m6A demethylases FTO and ALKBH5, which was associated with reduced global and SLC10A2-specific m6A modification. In vitro experiments in IPEC-J2 cells suggested that inhibition of FTO or glutamate dehydrogenase attenuated the effects of glutamate on m6A modification and SLC10A2 expression, and that YTHDF2 may contribute to the degradation of hyper-m6A-modified SLC10A2 mRNA. Glutamate-derived \u03b1-KG may serve as a cofactor supporting demethylase expression and activity after LPS stimulation. Collectively, these findings suggest that glutamate may alleviate LPS-induced intestinal inflammation in weaned piglets through a pathway involving \u03b1-KG-dependent maintenance of m6A demethylase expression, reduced m6A modification of SLC10A2, and preserved ASBT\/FXR signaling. This study provides evidence for a nutritional\u2013epitranscriptomic axis that may contribute to intestinal protection under acute inflammatory stress.<\/jats:p>","DOI":"10.3390\/ani16182854","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T14:11:22Z","timestamp":1789049482000},"page":"2854","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Dietary Supplementation of Glutamate Alleviates LPS-Induced Intestinal Injury Associated with m6A Modification of SLC10A2 in Weaned Piglets"],"prefix":"10.3390","volume":"16","author":[{"given":"Zhending","family":"Gan","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiawei","family":"He","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiyue","family":"Jin","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiuqin","family":"Ma","sequence":"additional","affiliation":[{"name":"Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Crop and Livestock Integration, Ministry of Agriculture and Rural Affairs, Nanjing 210014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanlong","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Yangzhou University, Yangzhou 225000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8622-8078","authenticated-orcid":false,"given":"Xiang","family":"Zhong","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"Institute of Animal Nutrition and Health Industry, Anyou Biotechnology, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"Natural Plant and Animal Health Innovation Institute, NJAU-Cohoo Biotechnology, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"State Key Laboratory of Meat Quality Control and Cultured Meat Development, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-3-030-45328-2_1","article-title":"Amino Acids in Intestinal Physiology and Health","volume":"1265","author":"Beaumont","year":"2020","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.aninu.2023.07.009","article-title":"Amino acids in piglet diarrhea: Effects, mechanisms and insights","volume":"16","author":"Zhou","year":"2024","journal-title":"Anim. Nutr."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.molcel.2019.07.007","article-title":"\u03b1-Ketoglutarate-Activated NF-\u03baB Signaling Promotes Compensatory Glucose Uptake and Brain Tumor Development","volume":"76","author":"Wang","year":"2019","journal-title":"Mol. Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1146\/annurev-animal-022513-114113","article-title":"Amino acid nutrition in animals: Protein synthesis and beyond","volume":"2","author":"Wu","year":"2014","journal-title":"Annu. Rev. Anim. Biosci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2727","DOI":"10.1017\/S1751731119001800","article-title":"Review: The roles and functions of glutamine on intestinal health and performance of weaning pigs","volume":"13","author":"Ji","year":"2019","journal-title":"Anim. Int. J. Anim. Biosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2405","DOI":"10.1016\/j.scib.2024.03.055","article-title":"Excitatory amino acid transporter supports inflammatory macrophage responses","volume":"69","author":"Gan","year":"2024","journal-title":"Sci. Bull."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1038\/s41423-025-01375-7","article-title":"Excitatory amino acid transporters support mast cell degranulation via \u03b1-KG-mediated methylation of Spp1","volume":"23","author":"Gan","year":"2026","journal-title":"Cell. Mol. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"112597","DOI":"10.1016\/j.foodres.2023.112597","article-title":"Dietary glutamate enhances intestinal immunity by modulating microbiota and Th17\/Treg balance-related immune signaling in piglets after lipopolysaccharide challenge","volume":"166","author":"Liu","year":"2023","journal-title":"Food Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.aninu.2022.03.001","article-title":"Glutamine in suppression of lipopolysaccharide-induced piglet intestinal inflammation: The crosstalk between AMPK activation and mitochondrial function","volume":"10","author":"He","year":"2022","journal-title":"Anim. Nutr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1016\/j.tig.2016.09.006","article-title":"Update: Mechanisms Underlying N6-Methyladenosine Modification of Eukaryotic mRNA","volume":"32","author":"Wang","year":"2016","journal-title":"Trends Genet. TIG"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1038\/nrm3785","article-title":"The dynamic epitranscriptome: N6-methyladenosine and gene expression control","volume":"15","author":"Meyer","year":"2014","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1111\/imm.13574","article-title":"Ectopic endometriotic stromal cells-derived lactate induces M2 macrophage polarization via Mettl3\/Trib1\/ERK\/STAT3 signalling pathway in endometriosis","volume":"168","author":"Gou","year":"2023","journal-title":"Immunology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"C762","DOI":"10.1152\/ajpcell.00212.2019","article-title":"The N6-methyladenosine (m6A)-forming enzyme METTL3 facilitates M1 macrophage polarization through the methylation of STAT1 mRNA","volume":"317","author":"Liu","year":"2019","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"101338","DOI":"10.1016\/j.mam.2025.101338","article-title":"Metabolic reprogramming shapes post-translational modification in macrophages","volume":"102","author":"Han","year":"2025","journal-title":"Mol. Asp. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"17438","DOI":"10.1021\/acsomega.9b02236","article-title":"Resveratrol and Curcumin Improve Intestinal Mucosal Integrity and Decrease m6A RNA Methylation in the Intestine of Weaning Piglets","volume":"4","author":"Gan","year":"2019","journal-title":"ACS Omega"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1002\/lipd.12023","article-title":"Curcumin Attenuates Lipopolysaccharide-Induced Hepatic Lipid Metabolism Disorder by Modification of m6 A RNA Methylation in Piglets","volume":"53","author":"Lu","year":"2018","journal-title":"Lipids"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Xu, M., Zhuo, R., Tao, S., Liang, Y., Liu, C., Liu, Q., Wang, T., and Zhong, X. (2022). M6A RNA Methylation Mediates NOD1\/NF-kB Signaling Activation in the Liver of Piglets Challenged with Lipopolysaccharide. Antioxidants, 11.","DOI":"10.3390\/antiox11101954"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4747","DOI":"10.1007\/s11033-021-06421-x","article-title":"Demethyltransferase AlkBH1 substrate diversity and relationship to human diseases","volume":"48","author":"Zhang","year":"2021","journal-title":"Mol. Biol. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113163","DOI":"10.1016\/j.celrep.2023.113163","article-title":"ALKBH5 modulates hematopoietic stem and progenitor cell energy metabolism through m6A modification-mediated RNA stability control","volume":"42","author":"Gao","year":"2023","journal-title":"Cell Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.immuni.2015.02.005","article-title":"Network integration of parallel metabolic and transcriptional data reveals metabolic modules that regulate macrophage polarization","volume":"42","author":"Jha","year":"2015","journal-title":"Immunity"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1038\/s41423-024-01149-7","article-title":"Targeting macrophage metabolism to enhance tumor immunotherapy","volume":"21","author":"Liu","year":"2024","journal-title":"Cell. Mol. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6016","DOI":"10.1111\/febs.70119","article-title":"Glutamine limits NLRP3 inflammasome activation and pyroptosis in macrophages by sustaining the IRG1\/itaconate axis","volume":"292","author":"Chen","year":"2025","journal-title":"FEBS J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1038\/nprot.2012.148","article-title":"Transcriptome-wide mapping of N6-methyladenosine by m6A-seq based on immunocapturing and massively parallel sequencing","volume":"8","author":"Dominissini","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1816","DOI":"10.1016\/j.celrep.2018.10.068","article-title":"Circadian Clock Regulation of Hepatic Lipid Metabolism by Modulation of m6A mRNA Methylation","volume":"25","author":"Zhong","year":"2018","journal-title":"Cell Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e2215921119","DOI":"10.1073\/pnas.2215921119","article-title":"GABA signaling enforces intestinal germinal center B cell differentiation","volume":"119","author":"Liao","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"111650","DOI":"10.1016\/j.mce.2022.111650","article-title":"Immunomodulatory functions of FXR","volume":"551","author":"Fiorucci","year":"2022","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1111\/nyas.70014","article-title":"Bile acid modulation of host immunity","volume":"1551","author":"Yang","year":"2025","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1038\/nature12730","article-title":"N6-methyladenosine-dependent regulation of messenger RNA stability","volume":"505","author":"Wang","year":"2014","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"eabe9274","DOI":"10.1126\/sciadv.abe9274","article-title":"GABA transporter sustains IL-1\u03b2 production in macrophages","volume":"7","author":"Xia","year":"2021","journal-title":"Sci. Adv."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"111770","DOI":"10.1016\/j.celrep.2022.111770","article-title":"GABA regulates IL-1\u03b2 production in macrophages","volume":"41","author":"Fu","year":"2022","journal-title":"Cell Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2467213","DOI":"10.1080\/19490976.2025.2467213","article-title":"Interplay of m6A RNA methylation and gut microbiota in modulating gut injury","volume":"17","author":"Wang","year":"2025","journal-title":"Gut Microbes"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"eabl5723","DOI":"10.1126\/sciadv.abl5723","article-title":"m6A mRNA modification maintains colonic epithelial cell homeostasis via NF-\u03baB\u2013mediated antiapoptotic pathway","volume":"8","author":"Zhang","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"194402","DOI":"10.1016\/j.bbagrm.2019.07.003","article-title":"Small changes, big implications: The impact of m6A RNA methylation on gene expression in pluripotency and development","volume":"1862","author":"Heck","year":"2019","journal-title":"Biochim. Biophys. Acta Gene Regul. Mech."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"533","DOI":"10.5534\/wjmh.250125","article-title":"N6-Methyladenosine RNA Methylation: A Key Regulator in Male Reproductive Disorders and Urological Cancers","volume":"44","author":"Huang","year":"2026","journal-title":"World J. Men\u2019s Health"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e70078","DOI":"10.1002\/biof.70078","article-title":"Bile Acids as Key Mediators of the Gut Microbiota\u2013Immune Axis: Potential Biomarker and Therapeutic Perspectives","volume":"52","author":"Baldi","year":"2026","journal-title":"Biofactors"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Yang, Y., Ren, L., Zhang, Y., Wang, X., Shang, J., and Zhang, L. (2026). Microbiota in cholestatic diseases: Crosstalk among bile composition, the biliary microbiome, and host immunity. Front. Immunol., 17.","DOI":"10.3389\/fimmu.2026.1884030"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Du, J., Xiang, X., Xu, D., Zhang, J., Fang, W., Xu, W., Mai, K., and Ai, Q. (2021). FXR, a Key Regulator of Lipid Metabolism, Is Inhibited by ER Stress-Mediated Activation of JNK and p38 MAPK in Large Yellow Croakers (Larimichthys crocea) Fed High Fat Diets. Nutrients, 13.","DOI":"10.3390\/nu13124343"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1521","DOI":"10.1007\/s10695-022-01130-z","article-title":"Farnesoid X receptor regulates PI3K\/AKT\/mTOR signaling pathway, lipid metabolism, and immune response in hybrid grouper","volume":"48","author":"Xu","year":"2022","journal-title":"Fish Physiol. Biochem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"G829","DOI":"10.1152\/ajpgi.00155.2013","article-title":"Mechanisms of STAT3 activation in the liver of FXR knockout mice","volume":"305","author":"Li","year":"2013","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/18\/2854\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T04:13:13Z","timestamp":1789445593000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/18\/2854"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,10]]},"references-count":39,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ani16182854"],"URL":"https:\/\/doi.org\/10.3390\/ani16182854","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,10]]}}}