{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T14:45:40Z","timestamp":1790952340127,"version":"4.1.0"},"reference-count":51,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2024,10,21]],"date-time":"2024-10-21T00:00:00Z","timestamp":1729468800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Endothelial dysfunction, which is marked by a reduction in nitric oxide (NO) production or an imbalance in relaxing and contracting factor levels, exacerbates atherosclerosis by promoting the production of cell adhesion molecules and cytokines. This study aimed to investigate the effects of Limosilactobacillus reuteri HY7503, a novel probiotic isolated from raw milk, on endothelial dysfunction. Five lactic acid bacterial strains were screened for their antioxidant, anti-inflammatory, and endothelium-protective properties; L. reuteri HY7503 had the most potent effect. In a mouse model of angiotensin II-induced endothelial dysfunction, L. reuteri HY7503 reduced vascular thickening (19.78%), increased serum NO levels (226.70%), upregulated endothelial NO synthase (eNOS) expression in the aortic tissue, and decreased levels of cell adhesion molecules (intercellular adhesion molecule-1 [ICAM-1] and vascular cell adhesion molecule-1 [VCAM-1]) and serum cytokines (tumor necrosis factor-alpha [TNF-\u03b1] and interleukin-6 [IL-6]). In TNF-\u03b1-treated human umbilical vein endothelial cells (HUVECs), L. reuteri HY7503 enhanced NO production and reduced cell adhesion molecule levels. In HUVECs, surface-layer proteins (SLPs) were more effective than extracellular vesicles (exosomes) in increasing NO production and decreasing cell adhesion molecule levels. These findings suggested that L. reuteri HY7503 may serve as a functional probiotic that alleviates endothelial dysfunction.<\/jats:p>","DOI":"10.3390\/ijms252011326","type":"journal-article","created":{"date-parts":[[2024,10,21]],"date-time":"2024-10-21T10:49:22Z","timestamp":1729507762000},"page":"11326","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Limosilactobacillus reuteri HY7503 and Its Cellular Proteins Alleviate Endothelial Dysfunction by Increasing Nitric Oxide Production and Regulating Cell Adhesion Molecule Levels"],"prefix":"10.3390","volume":"25","author":[{"given":"Hyejin","family":"Jeon","sequence":"first","affiliation":[{"name":"R&BD Center, Hy Co., Ltd., 22 Giheungdanji-ro 24 Beon-gil, Giheung-gu, Yongin-si 17086, Gyeonggi-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8258-119X","authenticated-orcid":false,"given":"Daehyeop","family":"Lee","sequence":"additional","affiliation":[{"name":"R&BD Center, Hy Co., Ltd., 22 Giheungdanji-ro 24 Beon-gil, Giheung-gu, Yongin-si 17086, Gyeonggi-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5534-2871","authenticated-orcid":false,"given":"Joo-Yun","family":"Kim","sequence":"additional","affiliation":[{"name":"R&BD Center, Hy Co., Ltd., 22 Giheungdanji-ro 24 Beon-gil, Giheung-gu, Yongin-si 17086, Gyeonggi-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jae-Jung","family":"Shim","sequence":"additional","affiliation":[{"name":"R&BD Center, Hy Co., Ltd., 22 Giheungdanji-ro 24 Beon-gil, Giheung-gu, Yongin-si 17086, Gyeonggi-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jae-Hwan","family":"Lee","sequence":"additional","affiliation":[{"name":"R&BD Center, Hy Co., Ltd., 22 Giheungdanji-ro 24 Beon-gil, Giheung-gu, Yongin-si 17086, Gyeonggi-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/j.ccc.2019.12.009","article-title":"Nitric Oxide and Endothelial Dysfunction","volume":"36","author":"Cyr","year":"2020","journal-title":"Crit. Care Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1007\/s11906-003-0055-4","article-title":"Endothelial nitric oxide synthase and endothelial dysfunction","volume":"5","author":"Huang","year":"2003","journal-title":"Curr. Hypertens. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.tice.2018.09.002","article-title":"Alterations in intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) in human endothelial cells","volume":"54","author":"Habas","year":"2018","journal-title":"Tissue Cell"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Gomolak, J.R., and Didion, S.P. (2014). Angiotensin II-induced endothelial dysfunction is temporally linked with increases in interleukin-6 and vascular macrophage accumulation. Front. Physiol., 5.","DOI":"10.3389\/fphys.2014.00396"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"100084","DOI":"10.1016\/j.hsr.2023.100084","article-title":"Endothelial dysfunction and its relation in different disorders: Recent update","volume":"7","author":"Ray","year":"2023","journal-title":"Health Sci. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.3346\/jkms.2015.30.9.1213","article-title":"Endothelial dysfunction: Clinical implications in cardiovascular disease and therapeutic approaches","volume":"30","author":"Park","year":"2015","journal-title":"J. Korean Med. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1642","DOI":"10.1016\/j.foodchem.2016.10.124","article-title":"Molecular mechanisms and in vitro antioxidant effects of Lactobacillus plantarum MA2","volume":"221","author":"Tang","year":"2017","journal-title":"Food Chem."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Lin, X., Xia, Y., Wang, G., Yang, Y., Xiong, Z., Lv, F., Zhou, W., and Ai, L. (2018). Lactic acid bacteria with antioxidant activities alleviating oxidized oil induced hepatic injury in mice. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02684"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Mathur, H., Beresford, T.P., and Cotter, P.D. (2020). Health Benefits of Lactic Acid Bacteria (LAB) Fermentates. Nutrients, 12.","DOI":"10.3390\/nu12061679"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1223","DOI":"10.3168\/jds.2019-17092","article-title":"Characterization and evaluation of lactic acid bacteria from indigenous raw milk for potential probiotic properties","volume":"103","author":"Reuben","year":"2020","journal-title":"J. Dairy Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1186\/s12967-015-0759-7","article-title":"Chronic administration of the probiotic kefir improves the endothelial function in spontaneously hypertensive rats","volume":"13","author":"Friques","year":"2015","journal-title":"J. Transl. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1186\/s12934-020-1289-4","article-title":"Surface components and metabolites of probiotics for regulation of intestinal epithelial barrier","volume":"19","author":"Liu","year":"2020","journal-title":"Microb. Cell Factories"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Bajic, S.S., Ca\u00f1as, M.-A., Tolinacki, M., Badia, J., S\u00e1nchez, B., Golic, N., Margolles, A., Baldom\u00e1, L., and Ruas-Madiedo, P. (2020). Proteomic profile of extracellular vesicles released by Lactiplantibacillus plantarum BGAN8 and their internalization by non-polarized HT29 cell line. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-78920-z"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kaur, N., and Dey, P. (2023). Bacterial exopolysaccharides as emerging bioactive macromolecules: From fundamentals to applications. Res. Microbiol., 174.","DOI":"10.1016\/j.resmic.2022.104024"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5225","DOI":"10.1007\/s00253-013-4962-2","article-title":"Lactobacillus surface layer proteins: Structure, function and applications","volume":"97","author":"Palva","year":"2013","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/s00281-018-0680-2","article-title":"Extracellular vesicles and their immunomodulatory functions in pregnancy","volume":"40","author":"Nair","year":"2018","journal-title":"Semin. Immunopathol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Mata Forsberg, M., Bj\u00f6rkander, S., Pang, Y., Lundqvist, L., Ndi, M., Ott, M., Escrib\u00e1, I.B., Jaeger, M.C., Roos, S., and Sverremark-Ekstr\u00f6m, E. (2019). Extracellular membrane vesicles from lactobacilli dampen IFN-\u03b3 responses in a monocyte-dependent manner. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-53576-6"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"B\u00e4uerl, C., Coll-Marqu\u00e9s, J.M., Tarazona-Gonz\u00e1lez, C., and P\u00e9rez-Mart\u00ednez, G. (2020). Lactobacillus casei extracellular vesicles stimulate EGFR pathway likely due to the presence of proteins P40 and P75 bound to their surface. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-75930-9"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1136\/heart.85.3.342","article-title":"Endothelial function and nitric oxide: Clinical relevance","volume":"85","author":"Vallance","year":"2001","journal-title":"Heart"},{"key":"ref_20","first-page":"183","article-title":"Endothelial dysfunction: Cardiovascular risk factors, therapy, and outcome","volume":"1","author":"Hadi","year":"2005","journal-title":"Vasc. Health Risk Manag."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"719","DOI":"10.4330\/wjc.v7.i11.719","article-title":"Vascular endothelial dysfunction and pharmacological treatment","volume":"7","author":"Su","year":"2015","journal-title":"World J. Cardiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.ijcard.2019.11.004","article-title":"Improvement in endothelial function in cardiovascular disease\u2014Is arginase the target?","volume":"301","author":"Mahdi","year":"2020","journal-title":"Int. J. Cardiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1016\/j.hlc.2021.09.006","article-title":"The Effects of Probiotics on Inflammation, Endothelial Dysfunction, and Atherosclerosis Progression: A Mechanistic Overview","volume":"31","author":"Salari","year":"2022","journal-title":"Heart Lung Circ."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"10792","DOI":"10.3168\/jds.2018-14989","article-title":"Assessing and comparing antioxidant activities of lactobacilli strains by using different chemical and cellular antioxidant methods","volume":"101","author":"Mu","year":"2018","journal-title":"J. Dairy Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1038\/nature07201","article-title":"Origin and physiological roles of inflammation","volume":"454","author":"Medzhitov","year":"2008","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7121","DOI":"10.4049\/jimmunol.179.10.7121","article-title":"Apigenin blocks lipopolysaccharide-induced lethality in vivo and proinflammatory cytokines expression by inactivating NF-\u03baB through the suppression of p65 phosphorylation","volume":"179","author":"Nicholas","year":"2007","journal-title":"J. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.jss.2007.12.758","article-title":"Lipopolysaccharide-Stimulated RAW 264.7 Macrophage Inducible Nitric Oxide Synthase and Nitric Oxide Production Is Decreased by an Omega-3 Fatty Acid Lipid Emulsion","volume":"149","author":"Aldridge","year":"2008","journal-title":"J. Surg. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1420","DOI":"10.1038\/cdd.2010.19","article-title":"TNF-\u03b1-induced ROS production triggering apoptosis is directly linked to Romo1 and Bcl-XL","volume":"17","author":"Kim","year":"2010","journal-title":"Cell Death Differ."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1002\/jbm.a.35233","article-title":"Nitric oxide secretion by endothelial cells in response to fluid shear stress, aspirin, and temperature","volume":"103","author":"Kabirian","year":"2014","journal-title":"J. Biomed. Mater. Res. Part A"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1146\/annurev.pharmtox.44.101802.121844","article-title":"The regulation and pharmacology of endothelial nitric oxide synthase","volume":"46","author":"Dudzinski","year":"2006","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"C1181","DOI":"10.1152\/ajpcell.00328.2010","article-title":"Angiotensin II-induced vascular endothelial dysfunction through RhoA\/Rho kinase\/p38 mitogen-activated protein kinase\/arginase pathway","volume":"300","author":"Shatanawi","year":"2011","journal-title":"Am. J. Physiol. Physiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1016\/S0008-6363(96)00213-1","article-title":"The influence of angiotensin II-induced increase in aortic wall mass on compliance in rats in vivo","volume":"33","author":"Smits","year":"1997","journal-title":"Cardiovasc. Res."},{"key":"ref_33","first-page":"E867","article-title":"Mechanisms contributing to angiotensin II regulation of body weight","volume":"274","author":"Cassis","year":"1998","journal-title":"Am. J. Physiol. Metab."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"H1932","DOI":"10.1152\/ajpheart.00961.2009","article-title":"Weight loss in obese C57BL\/6 mice limits adventitial expansion of established angiotensin II-induced abdominal aortic aneurysms","volume":"298","author":"Police","year":"2010","journal-title":"Am. J. Physiol. Circ. Physiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.jpeds.2009.08.036","article-title":"Aortic Intima Media Thickness is an Early Marker of Atherosclerosis in Children with Type 1 Diabetes Mellitus","volume":"156","author":"Harrington","year":"2010","journal-title":"J. Pediatr."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"183","DOI":"10.5114\/amsad.2019.87002","article-title":"Aortic intima-media thickness can be used to determine target organ damage in adult patients with coronary artery disease risk factors","volume":"4","author":"Bulut","year":"2019","journal-title":"Arch. Med. Sci.-Atheroscler. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1298","DOI":"10.1016\/j.freeradbiomed.2009.02.011","article-title":"Antioxidant treatment reduces matrix metalloproteinase-2-induced vascular changes in renovascular hypertension","volume":"46","author":"Castro","year":"2009","journal-title":"Free. Radic. Biol. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1007\/s00210-012-0798-z","article-title":"Euterpe oleracea mart.-derived polyphenols prevent endothelial dysfunction and vascular structural changes in renovascular hypertensive rats: Role of oxidative stress","volume":"385","author":"Ognibene","year":"2012","journal-title":"Naunyn-Schmiedebergs Arch. Pharmacol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"155","DOI":"10.34297\/AJBSR.2022.15.002087","article-title":"Endothelial Nitric Oxide Synthase (eNOS) and the Cardiovascular System: In Physiology and in Disease States","volume":"15","author":"Tran","year":"2022","journal-title":"Am. J. Biomed. Sci. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.semnephrol.2004.04.008","article-title":"Nitric oxide, angiotensin II, and hypertension","volume":"24","author":"Zhou","year":"2004","journal-title":"Semin. Nephrol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1161\/01.ATV.0000125114.88079.96","article-title":"Dysfunction of Endothelial Nitric Oxide Synthase and Atherosclerosis","volume":"24","author":"Kawashima","year":"2004","journal-title":"Arter. Thromb. Vasc. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"104802","DOI":"10.1016\/j.amsu.2022.104802","article-title":"Novel insights on the role of VCAM-1 and ICAM-1: Potential biomarkers for cardiovascular diseases","volume":"84","author":"Kaur","year":"2022","journal-title":"Ann. Med. Surg."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Trejo-Moreno, C., Jim\u00e9nez-Ferrer, E., Castro-Mart\u00ednez, G., M\u00e9ndez-Mart\u00ednez, M., Santana, M.A., Arrell\u00edn-Rosas, G., Pedraza-Chaverri, J., Medina-Campos, O.N., Hern\u00e1ndez-T\u00e9llez, B., and Ram\u00edrez-Pliego, O. (2021). Characterization of a murine model of endothelial dysfunction induced by chronic intraperitoneal administration of angiotensin II. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-00676-x"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1007\/s00395-008-0733-0","article-title":"The role of inflammatory cytokines in endothelial dysfunction","volume":"103","author":"Zhang","year":"2008","journal-title":"Basic. Res. Cardiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1186\/s13045-014-0080-6","article-title":"Immunosuppressive\/anti-inflammatory cytokines directly and indirectly inhibit endothelial dysfunction\u2014A novel mechanism for maintaining vascular function","volume":"7","author":"Shao","year":"2014","journal-title":"J. Hematol. Oncol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1099\/mic.0.000766","article-title":"Comparative analysis of immunological properties of S-layer proteins isolated from Lactobacillus strains","volume":"165","author":"Suzuki","year":"2019","journal-title":"Microbiology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"8344","DOI":"10.1128\/AEM.71.12.8344-8351.2005","article-title":"Functional analysis of putative adhesion factors in Lactobacillus acidophilus NCFM","volume":"71","author":"Buck","year":"2005","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"R125","DOI":"10.1093\/hmg\/dds317","article-title":"Exosomes and microvesicles: Extracellular vesicles for genetic information transfer and gene therapy","volume":"21","author":"Lee","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1038\/1811199a0","article-title":"Antioxidant Determinations by the Use of a Stable Free Radical","volume":"181","author":"Blois","year":"1958","journal-title":"Nature"},{"key":"ref_50","unstructured":"Riss, T.L., Moravec, R.A., Niles, A.L., Duellman, S., Benink, H.A., Worzella, T.J., and Minor, L. (2024, October 01). Cell Viability Assays, Assay Guidance Manual [Internet], Available online: http:\/\/www.ncbi.nlm.nih.gov\/books\/NBK144065\/."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/S0076-6879(07)00823-3","article-title":"Nitrite and Nitrate Measurement by Griess Reagent in Human Plasma: Evaluation of Interferences and Standardization","volume":"440","author":"Giustarini","year":"2008","journal-title":"Methods Enzymol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/20\/11326\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:17:40Z","timestamp":1760113060000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/20\/11326"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,21]]},"references-count":51,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2024,10]]}},"alternative-id":["ijms252011326"],"URL":"https:\/\/doi.org\/10.3390\/ijms252011326","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,21]]}}}