{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T00:47:25Z","timestamp":1760402845842,"version":"build-2065373602"},"reference-count":37,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2020,4,17]],"date-time":"2020-04-17T00:00:00Z","timestamp":1587081600000},"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>Type 1 Diabetes Mellitus (T1D) is associated with accelerated atherosclerosis that is responsible for high morbidity and mortality. Endothelial hyperpermeability, a feature of endothelial dysfunction, is an early step of atherogenesis since it favours intimal lipid uptake. Therefore, we tested endothelial leakage by loading the sera from T1D patients onto cultured human endothelial cells and found it increased by hyperglycaemic sera. These results were phenocopied in endothelial cells cultured in a medium containing high concentrations of glucose, which activates inducible nitric oxide synthase with a consequent increase of nitric oxide. Inhibition of the enzyme prevented high glucose-induced hyperpermeability, thus pointing to nitric oxide as the mediator involved in altering the endothelial barrier function. Since nitric oxide is much higher in sera from hyperglycaemic than normoglycaemic T1D patients, and the inhibition of inducible nitric oxide synthase prevents sera-dependent increased endothelial permeability, this enzyme might represent a promising biochemical marker to be monitored in T1D patients to predict alterations of the vascular wall, eventually promoting intimal lipid accumulation.<\/jats:p>","DOI":"10.3390\/ijms21082798","type":"journal-article","created":{"date-parts":[[2020,4,21]],"date-time":"2020-04-21T04:49:38Z","timestamp":1587444578000},"page":"2798","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Endothelial Hyper-Permeability Induced by T1D Sera Can be Reversed by iNOS Inactivation"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7153-2256","authenticated-orcid":false,"given":"Alessandra","family":"Cazzaniga","sequence":"first","affiliation":[{"name":"Department of Biomedical and Clinical Sciences \u201cLuigi Sacco\u201d, Universit\u00e0 di Milano, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8888-2594","authenticated-orcid":false,"given":"Roberta","family":"Scrimieri","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Clinical Sciences \u201cLuigi Sacco\u201d, Universit\u00e0 di Milano, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa","family":"Giani","sequence":"additional","affiliation":[{"name":"Humanitas Clinical and Research Center, 20089 Rozzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gian Vincenzo","family":"Zuccotti","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Clinical Sciences \u201cLuigi Sacco\u201d, Universit\u00e0 di Milano, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeanette A. M.","family":"Maier","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Clinical Sciences \u201cLuigi Sacco\u201d, Universit\u00e0 di Milano, 20157 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mcna.2014.08.015","article-title":"Update on Diabetes Classification","volume":"99","author":"Thomas","year":"2015","journal-title":"Med Clin. N. Am."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1002\/dmrr.1195","article-title":"Macrovascular angiopathy in children and adolescents with type 1 diabetes","volume":"27","author":"Giannini","year":"2011","journal-title":"Diabetes\/Metab. Res. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"201","DOI":"10.2174\/1573399810666140613124326","article-title":"Vascular endothelial dysfunction and nutritional compounds in early type 1 diabetes","volume":"10","author":"Hoffman","year":"2014","journal-title":"Curr. Diabetes Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"620","DOI":"10.1161\/CIRCRESAHA.115.306301","article-title":"Endothelial Cell Dysfunction and the Pathobiology of Atherosclerosis","volume":"118","author":"Gimbrone","year":"2016","journal-title":"Circ. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1285","DOI":"10.1161\/CIRCULATIONAHA.106.652859","article-title":"Endothelial Function and Dysfunction","volume":"115","author":"Deanfield","year":"2007","journal-title":"Circulation"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1007\/s00441-008-0685-6","article-title":"Endothelial dysfunction and diabetes: Roles of hyperglycemia, impaired insulin signaling and obesity","volume":"335","author":"Bakker","year":"2008","journal-title":"Cell Tissue Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1111\/apa.12877","article-title":"V Adolescents and young adults with type 1 diabetes display a high prevalence of endothelial dysfunction","volume":"104","author":"Scaramuzza","year":"2015","journal-title":"Acta Paediatr."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1172\/JCI66843","article-title":"Linking endothelial dysfunction with endothelial cell activation","volume":"123","author":"Liao","year":"2013","journal-title":"J. Clin. Investig."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.ijdm.2010.09.002","article-title":"Endothelial cell dysfunction in hyperglycemia: Phenotypic change, intracellular signaling modification, ultrastructural alteration, and potential clinical outcomes","volume":"2","author":"Popov","year":"2010","journal-title":"Int. J. Diabetes Mellit."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1007\/s10787-007-0013-x","article-title":"Role of nitric oxide in inflammatory diseases","volume":"15","author":"Sharma","year":"2007","journal-title":"Inflammopharmacology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.dsx.2012.09.012","article-title":"Evaluation of nitric oxide metabolites in a group of subjects with metabolic syndrome","volume":"6","author":"Caimi","year":"2012","journal-title":"Diabetes Metab. Syndr. Clin. Res. Rev."},{"key":"ref_12","first-page":"5541","article-title":"eNOS-derived nitric oxide regulates endothelial barrier function through VE-cadherin and Rho GTPases","volume":"126","author":"Lin","year":"2013","journal-title":"J. Cell Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1620","DOI":"10.1111\/bph.12832","article-title":"Measurement of NO in biological samples","volume":"172","author":"Csonka","year":"2014","journal-title":"Br. J. Pharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.2337\/diabetes.49.7.1258","article-title":"Increased circulating nitric oxide in young patients with type 1 diabetes and persistent microalbuminuria: Relation to glomerular hyperfiltration","volume":"49","author":"Chiarelli","year":"2000","journal-title":"Diabetes"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Gonz\u00e1lez, M., Rojas, S., Avila, P., Cabrera, L., Villalobos, R., Palma, C., Aguayo, C., Pe\u00f1a, E., Gallardo, V., and Guzm\u00e1n-Guti\u00e9rrez, E. (2015). Insulin Reverses D-Glucose\u2013Increased Nitric Oxide and Reactive Oxygen Species Generation in Human Umbilical Vein Endothelial Cells. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0122398"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Adela, R., Nethi, S.K., Bagul, P.K., Barui, A.K., Mattapally, S., Kuncha, M., Patra, C.R., Reddy, P.N.C., and Banerjee, S.K. (2015). Hyperglycaemia Enhances Nitric Oxide Production in Diabetes: A Study from South Indian Patients. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0125270"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.niox.2016.09.009","article-title":"Nitric oxide levels in patients with diabetes mellitus: A systematic review and meta-analysis","volume":"61","author":"Assmann","year":"2016","journal-title":"Nitric Oxide"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.tem.2019.10.001","article-title":"Role of Nitric Oxide in Insulin Secretion and Glucose Metabolism","volume":"31","author":"Bahadoran","year":"2019","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1006\/niox.2000.0319","article-title":"A Rapid, Simple Spectrophotometric Method for Simultaneous Detection of Nitrate and Nitrite","volume":"5","author":"Miranda","year":"2001","journal-title":"Nitric Oxide"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.cmet.2012.11.012","article-title":"Vascular complications of diabetes: Mechanisms of injury and protective factors","volume":"17","author":"King","year":"2013","journal-title":"Cell Metab."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1186\/s13098-015-0098-0","article-title":"Effects of high glucose on human umbilical vein endothelial cell permeability and myosin light chain phosphorylation","volume":"7","author":"Zhao","year":"2015","journal-title":"Diabetol. Metab. Syndr."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Yang, B., Cai, B., Deng, P., Wu, X., Guan, Y., Zhang, B., Cai, W., Schaper, J., and Schaper, W. (2015). Nitric Oxide Increases Arterial Endotheial Permeability through Mediating VE-Cadherin Expression during Arteriogenesis. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127931"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Testa, I., Bonfigli, A.R., Prattichizzo, F., La Sala, L., De Nigris, V., and Ceriello, A. (2017). The \u201cMetabolic Memory\u201d Theory and the Early Treatment of Hyperglycemia in Prevention of Diabetic Complications. Nutrients, 9.","DOI":"10.3390\/nu9050437"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1007\/s00125-011-2049-0","article-title":"Glucose oscillations, more than constant high glucose, induce p53 activation and a metabolic memory in human endothelial cells","volume":"54","author":"Schisano","year":"2011","journal-title":"Diabetology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/j.clinbiochem.2009.09.011","article-title":"Reference values for serum nitric oxide metabolites in an adult population","volume":"43","author":"Ghasemi","year":"2010","journal-title":"Clin. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1118","DOI":"10.1111\/pedi.12913","article-title":"Associations between circulating inflammatory markers, diabetes type and complications in youth","volume":"20","author":"Aulich","year":"2019","journal-title":"Pediatr. Diabetes"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Fatima, N., Faisal, S.M., Zubair, S., Ajmal, M., Siddiqui, S.S., Moin, S., and Owais, M. (2016). Role of Pro-Inflammatory Cytokines and Biochemical Markers in the Pathogenesis of Type 1 Diabetes: Correlation with Age and Glycemic Condition in Diabetic Human Subjects. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0161548"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"361","DOI":"10.3892\/etm.2013.1166","article-title":"High glucose enhances LPS-stimulated human PMVEC hyperpermeability via the NO pathway","volume":"6","author":"Liu","year":"2013","journal-title":"Exp. Ther. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.mvr.2005.11.003","article-title":"Albumin leak across human pulmonary microvascular vs. umbilical vein endothelial cells under septic conditions","volume":"71","author":"Shelton","year":"2006","journal-title":"Microvasc. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/j.lfs.2018.11.030","article-title":"Involvement of inducible nitric oxide synthase and estrogen receptor ESR2 (ERbeta) in the vascular dysfunction in female type 1 diabetic rats","volume":"216","author":"Sartoretto","year":"2019","journal-title":"Life Sci."},{"key":"ref_31","first-page":"4205","article-title":"Effects of aerobic exercise on the expressions and activities of nitric oxide synthases in the blood vessel endothelium in prediabetes mellitus","volume":"17","author":"Wang","year":"2019","journal-title":"Exp. Ther. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2036","DOI":"10.1080\/10408398.2013.811212","article-title":"Dietary Nitrate, Nitric Oxide, and Cardiovascular Health","volume":"56","author":"Bondonno","year":"2015","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"McCarthy, O., Moser, O., Eckstein, M.L., Bain, S., Pitt, J., and Bracken, R.M. (2019). Supplementary Nitric Oxide Donors and Exercise as Potential Means to Improve Vascular Health in People with Type 1 Diabetes: Yes to NO?. Nutrients, 11.","DOI":"10.3390\/nu11071571"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5957","DOI":"10.1096\/fj.201801586RR","article-title":"The dynamic adaptation of primary human endothelial cells to simulated microgravity","volume":"33","author":"Cazzaniga","year":"2019","journal-title":"FASEB J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.1016\/j.jnutbio.2011.06.016","article-title":"Nitric oxide mediates low magnesium inhibition of osteoblast-like cell proliferation","volume":"23","author":"Leidi","year":"2012","journal-title":"J. Nutr. Biochem."},{"key":"ref_36","first-page":"16","article-title":"Magnesium and the blood-brain barrier in vitro: Effects on permeability and magnesium transport","volume":"32","author":"Romeo","year":"2019","journal-title":"Magnes. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"654","DOI":"10.1016\/j.ejcb.2010.05.001","article-title":"EDF-1 contributes to the regulation of nitric oxide release in VEGF-treated human endothelial cells","volume":"89","author":"Leidi","year":"2010","journal-title":"Eur. J. Cell Boil."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/8\/2798\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:08:57Z","timestamp":1760364537000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/8\/2798"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,17]]},"references-count":37,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["ijms21082798"],"URL":"https:\/\/doi.org\/10.3390\/ijms21082798","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2020,4,17]]}}}