{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T18:00:34Z","timestamp":1785261634179,"version":"3.55.0"},"reference-count":29,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Scientific Research Project of Tianjin Municipal Education Commission","award":["2022KJ278"],"award-info":[{"award-number":["2022KJ278"]}]},{"name":"Tianjin Key Medical Discipline Construction Project","award":["No.TJYXZDXK-3-014B"],"award-info":[{"award-number":["No.TJYXZDXK-3-014B"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>This study aims to examine whether oral glutamate administration is associated with accelerated atherosclerosis progression in high-fat diet-fed ApoE\u2009\u207b\u2009\/\u2009\u207b\u2009mice and to identify candidate protein associated with this process.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Atherosclerosis (AS) was induced in ApoE\u2009\u207b\u2009\/\u2009\u207b\u2009mice by feeding a high-fat diet (HFD) for three months. The mice were randomly divided into four groups: (1) the AS-GLU group, receiving glutamate with HFD; (2) the AS group, receiving HFD with placebo; (3) the Glu group, receiving glutamate with normal chow; and (4) the Control group, receiving normal chow with a placebo. Atherosclerotic lesions were assessed using\u00a0Lillie's\u00a0oil red O staining and histological analysis. LC-MS\/MS based proteomics profiling was employed to identify differentially expressed proteins, with subsequent validation by Western blotting and qPCR.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Glutamate administration alone (Glu group) induced minimal plaque formation. The combination of HFD and glutamate (AS-GLU) was associated with a greater atherosclerotic plaque area compared to HFD alone (AS group). Proteomics identified RAPGEF1, OPN, and MYL7 as proteins whose expression was increased in the AS-GLU group compared to AS group. These associations were confirmed by Western blot and qPCR.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This study shows that oral glutamate administration, in the context of a high-fat diet in ApoE\u2009\u207b\u2009\/\u2009\u207b\u2009mice, is associated with exacerbated atherosclerosis and increased expression of RAPGEF1, OPN, and MYL7. These proteins represent candidate molecules associated with glutamate exposure, but their causal role remains to be determined. Serum glutamate levels were not significantly elevated, and lipid profile changes may also contribute to the observed associations.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0354719","type":"journal-article","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T17:24:06Z","timestamp":1785259446000},"page":"e0354719","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Glutamate administration is associated with aggravated atherosclerosis and altered expression of RAPGEF1, OPN, and MYL7 in High-Fat Diet-Fed ApoE\u207b\/\u207b Mice"],"prefix":"10.1371","volume":"21","author":[{"given":"Xiuli","family":"Cheng","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-0661-4886","authenticated-orcid":true,"given":"Fanshu","family":"Dai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dilraba","family":"Mahmut","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xingya","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9738-1123","authenticated-orcid":true,"given":"Biao","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"issue":"7855","key":"pone.0354719.ref001","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1038\/s41586-021-03392-8","article-title":"The changing landscape of atherosclerosis","volume":"592","author":"P Libby","year":"2021","journal-title":"Nature"},{"issue":"8","key":"pone.0354719.ref002","doi-asserted-by":"crossref","DOI":"10.1161\/CIR.0000000000000950","article-title":"Heart disease and stroke statistics-2021 update: a report from the American Heart Association","volume":"143","author":"SS Virani","year":"2021","journal-title":"Circulation"},{"issue":"2","key":"pone.0354719.ref003","doi-asserted-by":"crossref","DOI":"10.1210\/clinem\/dgaa898","article-title":"Elevated Circulating Glutamate Is Associated With Subclinical Atherosclerosis Independently of Established Risk Markers: A Cross-Sectional Study","volume":"106","author":"A Lehn-Stefan","year":"2021","journal-title":"J Clin Endocrinol Metab"},{"issue":"5","key":"pone.0354719.ref004","doi-asserted-by":"crossref","DOI":"10.1152\/ajpcell.00386.2005","article-title":"Glutamate induces oxidative stress and apoptosis in cerebral vascular endothelial cells: contributions of HO-1 and HO-2 to cytoprotection","volume":"290","author":"H Parfenova","year":"2006","journal-title":"Am J Physiol Cell Physiol"},{"issue":"9","key":"pone.0354719.ref005","doi-asserted-by":"crossref","DOI":"10.1161\/JAHA.116.003755","article-title":"Metabolites of glutamate metabolism are associated with incident cardiovascular events in the PREDIMED PREvenci\u00f3n con DIeta MEDiterr\u00e1nea (PREDIMED) Trial","volume":"5","author":"Y Zheng","year":"2016","journal-title":"J Am Heart Assoc"},{"issue":"10","key":"pone.0354719.ref006","doi-asserted-by":"crossref","first-page":"2206","DOI":"10.2337\/db20-0475","article-title":"Association of the 1q25 Diabetes-Specific Coronary Heart Disease Locus With Alterations of the \u03b3-Glutamyl Cycle and Increased Methylglyoxal Levels in Endothelial Cells","volume":"69","author":"C Pipino","year":"2020","journal-title":"Diabetes"},{"issue":"6","key":"pone.0354719.ref007","doi-asserted-by":"crossref","first-page":"1006","DOI":"10.1161\/01.ATV.0000128849.12617.f4","article-title":"Atherosclerosis in the apolipoprotein-E-deficient mouse: a decade of progress","volume":"24","author":"KS Meir","year":"2004","journal-title":"Arterioscler Thromb Vasc Biol"},{"issue":"1","key":"pone.0354719.ref008","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1002\/path.1749","article-title":"Proteomic analysis of differential protein expression in human atherosclerotic plaque progression","volume":"206","author":"MMPC Donners","year":"2005","journal-title":"J Pathol"},{"key":"pone.0354719.ref009","doi-asserted-by":"crossref","DOI":"10.1093\/nar\/gkae909","article-title":"KEGG: biological systems database as a model of the real world","volume":"53","author":"M Kanehisa","year":"2025","journal-title":"Nucleic Acids Res"},{"issue":"11","key":"pone.0354719.ref010","doi-asserted-by":"crossref","first-page":"1947","DOI":"10.1002\/pro.3715","article-title":"Toward understanding the origin and evolution of cellular organisms","volume":"28","author":"M Kanehisa","year":"2019","journal-title":"Protein Sci"},{"issue":"1","key":"pone.0354719.ref011","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1093\/nar\/28.1.27","article-title":"KEGG: kyoto encyclopedia of genes and genomes","volume":"28","author":"M Kanehisa","year":"2000","journal-title":"Nucleic Acids Res"},{"key":"pone.0354719.ref012","doi-asserted-by":"crossref","DOI":"10.1093\/nar\/gkq537","article-title":"The GeneMANIA prediction server: biological network integration for gene prioritization and predicting gene function","volume":"38","author":"D Warde-Farley","year":"2010","journal-title":"Nucleic Acids Res"},{"issue":"11","key":"pone.0354719.ref013","doi-asserted-by":"crossref","first-page":"2498","DOI":"10.1101\/gr.1239303","article-title":"Cytoscape: a software environment for integrated models of biomolecular interaction networks","volume":"13","author":"P Shannon","year":"2003","journal-title":"Genome Res"},{"key":"pone.0354719.ref014","article-title":"cytoHubba: identifying hub objects and sub-networks from complex interactome","author":"C-H Chin","year":"2014","journal-title":"BMC Syst Biol"},{"issue":"21","key":"pone.0354719.ref015","article-title":"GLS1-Mediated Redundancy in Glutamate Accelerates Arterial Calcification via Activating NMDAR\/Ca2+\/\u03b2-Catenin Pathway","volume":"12","author":"Z Zhou","year":"2025","journal-title":"Adv Sci (Weinh)"},{"key":"pone.0354719.ref016","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.jnutbio.2017.02.023","article-title":"Atherogenicity of amino acids in the lipid-laden macrophage model system in vitro and in atherosclerotic mice: a key role for triglyceride metabolism","volume":"45","author":"O Rom","year":"2017","journal-title":"J Nutr Biochem"},{"issue":"10","key":"pone.0354719.ref017","doi-asserted-by":"crossref","first-page":"1648","DOI":"10.1161\/01.ATV.14.10.1648","article-title":"Osteopontin is synthesized by macrophage, smooth muscle, and endothelial cells in primary and restenotic human coronary atherosclerotic plaques","volume":"14","author":"ER O\u2019Brien","year":"1994","journal-title":"Arterioscler Thromb"},{"issue":"3","key":"pone.0354719.ref018","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1007\/s11883-009-0032-8","article-title":"Osteopontin: a multifunctional protein at the crossroads of inflammation, atherosclerosis, and vascular calcification","volume":"11","author":"H-J Cho","year":"2009","journal-title":"Curr Atheroscler Rep"},{"issue":"5","key":"pone.0354719.ref019","doi-asserted-by":"crossref","first-page":"1194","DOI":"10.1002\/jcb.24462","article-title":"Osteopontin mediates macrophage chemotaxis via \u03b14 and \u03b19 integrins and survival via the \u03b14 integrin","volume":"114","author":"SA Lund","year":"2013","journal-title":"J Cell Biochem"},{"issue":"2","key":"pone.0354719.ref020","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1002\/path.6074","article-title":"Steroid hormone imbalance drives macrophage infiltration and Spp1\/osteopontin+ foam cell differentiation in the prostate","volume":"260","author":"P Popovics","year":"2023","journal-title":"J Pathol"},{"issue":"12","key":"pone.0354719.ref021","doi-asserted-by":"crossref","first-page":"2358","DOI":"10.1038\/s41401-023-01123-5","article-title":"Regulation of toll-like receptor (TLR) signaling pathways in atherosclerosis: from mechanisms to targeted therapeutics","volume":"44","author":"M Jin","year":"2023","journal-title":"Acta Pharmacol Sin"},{"issue":"6311","key":"pone.0354719.ref022","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1126\/science.aaf6659","article-title":"Senescent intimal foam cells are deleterious at all stages of atherosclerosis","volume":"354","author":"BG Childs","year":"2016","journal-title":"Science"},{"issue":"11","key":"pone.0354719.ref023","doi-asserted-by":"crossref","first-page":"3713","DOI":"10.1182\/blood.V93.11.3713","article-title":"CrkL activates integrin-mediated hematopoietic cell adhesion through the guanine nucleotide exchange factor C3G","volume":"93","author":"A Arai","year":"1999","journal-title":"Blood"},{"issue":"67","key":"pone.0354719.ref024","doi-asserted-by":"crossref","first-page":"110994","DOI":"10.18632\/oncotarget.22339","article-title":"C3G promotes a selective release of angiogenic factors from activated mouse platelets to regulate angiogenesis and tumor metastasis","volume":"8","author":"V Mart\u00edn-Granado","year":"2017","journal-title":"Oncotarget"},{"issue":"17","key":"pone.0354719.ref025","doi-asserted-by":"crossref","first-page":"2678","DOI":"10.1091\/mbc.e13-02-0098","article-title":"Rap-afadin axis in control of Rho signaling and endothelial barrier recovery","volume":"24","author":"AA Birukova","year":"2013","journal-title":"Mol Biol Cell"},{"issue":"12","key":"pone.0354719.ref026","doi-asserted-by":"crossref","DOI":"10.1096\/fj.202300830RR","article-title":"Rap1 small GTPase is essential for maintaining pulmonary endothelial barrier function in mice","volume":"37","author":"K Yamamoto","year":"2023","journal-title":"FASEB J"},{"issue":"5","key":"pone.0354719.ref027","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1291\/hypres.31.987","article-title":"Angiotensin II-induced osteopontin expression in vascular smooth muscle cells involves Gq\/11, Ras, ERK, Src and Ets-1","volume":"31","author":"K Abe","year":"2008","journal-title":"Hypertens Res"},{"issue":"5","key":"pone.0354719.ref028","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1159\/000516044","article-title":"Adapter Protein RapGEF1 Is Required for ERK1\/2 Signaling in Response to Elevated Phosphate in Vascular Smooth Muscle Cells","volume":"58","author":"NW Chavkin","year":"2021","journal-title":"J Vasc Res"},{"issue":"24","key":"pone.0354719.ref029","doi-asserted-by":"crossref","first-page":"6111","DOI":"10.1242\/dev.00831","article-title":"Embryonic atrial function is essential for mouse embryogenesis, cardiac morphogenesis and angiogenesis","volume":"130","author":"C Huang","year":"2003","journal-title":"Development"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0354719","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T17:24:12Z","timestamp":1785259452000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0354719"}},"subtitle":[],"editor":[{"given":"Hideto","family":"Sano","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,7,28]]},"references-count":29,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7,28]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0354719","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,28]]}}}