{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:38:52Z","timestamp":1790642332918,"version":"4.1.0"},"reference-count":39,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,10,30]],"date-time":"2018-10-30T00:00:00Z","timestamp":1540857600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003141","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda","doi-asserted-by":"publisher","award":["261915"],"award-info":[{"award-number":["261915"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Reverse cholesterol transport (RCT) is considered as the most important antiatherogenic role of high-density lipoproteins (HDL), but interventions based on RCT have failed to reduce the risk of coronary heart disease. In contrast to RCT, important evidence suggests that HDL deliver lipids to peripheral cells. Therefore, in this paper, we investigated whether HDL could improve endothelial function by delivering lipids to the cells. Internalization kinetics using cholesterol and apolipoprotein (apo) AI fluorescent double-labeled reconstituted HDL (rHDL), and human dermal microvascular endothelial cells-1 (HMEC-1) showed a fast cholesterol influx (10 min) and a slower HDL protein internalization as determined by confocal microscopy and flow cytometry. Sphingomyelin kinetics overlapped that of apo AI, indicating that only cholesterol became dissociated from rHDL during internalization. rHDL apo AI internalization was scavenger receptor class B type I (SR-BI)-dependent, whereas HDL cholesterol influx was independent of SR-BI and was not completely inhibited by the presence of low-density lipoproteins (LDL). HDL sphingomyelin was fundamental for intercellular adhesion molecule-1 (ICAM-1) downregulation in HMEC-1. However, vascular cell adhesion protein-1 (VCAM-1) was not inhibited by rHDL, suggesting that components such as apolipoproteins other than apo AI participate in HDL\u2019s regulation of this adhesion molecule. rHDL also induced endothelial nitric oxide synthase eNOS S1177 phosphorylation in HMEC-1 but only when the particle contained sphingomyelin. In conclusion, the internalization of HDL implies the dissociation of lipoprotein components and a SR-BI-independent fast delivery of cholesterol to endothelial cells. HDL internalization had functional implications that were mainly dependent on sphingomyelin. These results suggest a new role of HDL as lipid vectors to the cells, which could be congruent with the antiatherogenic properties of these lipoproteins.<\/jats:p>","DOI":"10.3390\/ijms19113394","type":"journal-article","created":{"date-parts":[[2018,10,31]],"date-time":"2018-10-31T03:50:56Z","timestamp":1540957856000},"page":"3394","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["HDL-Mediated Lipid Influx to Endothelial Cells Contributes to Regulating Intercellular Adhesion Molecule (ICAM)-1 Expression and eNOS Phosphorylation"],"prefix":"10.3390","volume":"19","author":[{"given":"M\u00f3nica","family":"Mu\u00f1oz-Vega","sequence":"first","affiliation":[{"name":"Molecular Biology Department, Instituto Nacional de Cardiolog\u00eda \u201cIgnacio Ch\u00e1vez\u201d, 14080 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felipe","family":"Mass\u00f3","sequence":"additional","affiliation":[{"name":"Physiology Departments, Instituto Nacional de Cardiolog\u00eda \u201cIgnacio Ch\u00e1vez\u201d, 14080 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Araceli","family":"P\u00e1ez","sequence":"additional","affiliation":[{"name":"Physiology Departments, Instituto Nacional de Cardiolog\u00eda \u201cIgnacio Ch\u00e1vez\u201d, 14080 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7916-5163","authenticated-orcid":false,"given":"Gilberto","family":"Vargas-Alarc\u00f3n","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Instituto Nacional de Cardiolog\u00eda \u201cIgnacio Ch\u00e1vez\u201d, 14080 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ram\u00f3n","family":"Coral-V\u00e1zquez","sequence":"additional","affiliation":[{"name":"Graduate School and Research Division, Escuela Superior de Medicina, Instituto Polit\u00e9cnico Nacional, 11340 M\u00e9xico City, Mexico"},{"name":"Sub-Directorate of Research and Education, Centro M\u00e9dico Nacional \u201c20 de Noviembre\u201d, Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado, 03100 M\u00e9xico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaime","family":"Mas-Oliva","sequence":"additional","affiliation":[{"name":"Instituto de Fisiolog\u00eda Celular, Universidad Nacional Aut\u00f3noma de M\u00e9xico, 04510 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3600-249X","authenticated-orcid":false,"given":"Elizabeth","family":"Carre\u00f3n-Torres","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Instituto Nacional de Cardiolog\u00eda \u201cIgnacio Ch\u00e1vez\u201d, 14080 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00d3scar","family":"P\u00e9rez-M\u00e9ndez","sequence":"additional","affiliation":[{"name":"Molecular Biology Department, Instituto Nacional de Cardiolog\u00eda \u201cIgnacio Ch\u00e1vez\u201d, 14080 Mexico City, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,10,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.plipres.2017.12.001","article-title":"Determinants of cholesterol efflux capacity in humans","volume":"69","author":"Talbot","year":"2018","journal-title":"Prog. Lipid Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.ccl.2017.12.013","article-title":"High-Density Lipoproteins: Effects on Vascular Function and Role in the Immune Response","volume":"36","author":"Haghikia","year":"2018","journal-title":"Cardiol. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.atherosclerosis.2018.06.881","article-title":"Endocytosis of lipoproteins","volume":"275","author":"Zanoni","year":"2018","journal-title":"Atherosclerosis"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"37457","DOI":"10.1074\/jbc.M605823200","article-title":"Role of scavenger receptor class B type I and sphingosine 1-phosphate receptors in high density lipoprotein-induced inhibition of adhesion molecule expression in endothelial cells","volume":"281","author":"Kimura","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"131","DOI":"10.4331\/wjbc.v4.i4.131","article-title":"High-density lipoprotein endocytosis in endothelial cells","volume":"4","author":"Fruhwurth","year":"2013","journal-title":"World J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1142","DOI":"10.1161\/CIRCRESAHA.108.190587","article-title":"High-density lipoprotein transport through aortic endothelial cells involves scavenger receptor BI and ATP-binding cassette transporter G1","volume":"104","author":"Rohrer","year":"2009","journal-title":"Circ. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"15886","DOI":"10.1038\/s41598-017-15949-7","article-title":"HDL particles incorporate into lipid bilayers\u2014A combined AFM and single molecule fluorescence microscopy study","volume":"7","author":"Plochberger","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1038\/89986","article-title":"High-density lipoprotein binding to scavenger receptor-BI activates endothelial nitric oxide synthase","volume":"7","author":"Yuhanna","year":"2001","journal-title":"Nat. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/S0022-2275(20)39816-3","article-title":"Regulation of rates of cholesterol synthesis in vivo in the liver and carcass of the rat measured using [3H]water","volume":"21","author":"Jeske","year":"1980","journal-title":"J. Lipid Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1161\/CIRCRESAHA.112.280081","article-title":"Scavenger receptor class B type I is a plasma membrane cholesterol sensor","volume":"112","author":"Saddar","year":"2013","journal-title":"Circ. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.1194\/jlr.M200173-JLR200","article-title":"The fate of HDL particles in vivo after SR-BI-mediated selective lipid uptake","volume":"43","author":"Webb","year":"2002","journal-title":"J. Lipid Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2172","DOI":"10.1093\/eurheartj\/ehy169","article-title":"Rare SCARB1 mutations associate with high-density lipoprotein cholesterol but not with coronary artery disease","volume":"39","author":"Helgadottir","year":"2018","journal-title":"Eur. Heart J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"7716","DOI":"10.3390\/ijms14047716","article-title":"Sphingomyelin in high-density lipoproteins: Structural role and biological function","volume":"14","author":"Osada","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.cca.2015.12.039","article-title":"HDL-sphingomyelin reduction after weight loss by an energy-restricted diet is associated with the improvement of lipid profile, blood pressure, and decrease of insulin resistance in overweight\/obese patients","volume":"454","author":"Madero","year":"2016","journal-title":"Clin. Chim. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1016\/j.clinbiochem.2015.03.019","article-title":"Possible involvement of sphingomyelin in the regulation of the plasma sphingosine 1-phosphate level in human subjects","volume":"48","author":"Ohkawa","year":"2015","journal-title":"Clin. Biochem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/978-1-4757-5806-1_5","article-title":"Sphingomyelin and cholesterol: From membrane biophysics and rafts to potential medical applications","volume":"37","author":"Barenholz","year":"2004","journal-title":"Subcell. Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1186\/s12944-018-0695-7","article-title":"Characterization of immortalized human dermal microvascular endothelial cells (HMEC-1) for the study of HDL functionality","volume":"17","author":"Masso","year":"2018","journal-title":"Lipids Health Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.bbalip.2015.11.004","article-title":"Itinerary of high density lipoproteins in endothelial cells","volume":"1861","author":"Perisa","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2260","DOI":"10.1161\/ATVBAHA.117.310290","article-title":"ABCA1-Derived Nascent High-Density Lipoprotein-Apolipoprotein AI and Lipids Metabolically Segregate","volume":"37","author":"Xu","year":"2017","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"15422","DOI":"10.1073\/pnas.222421399","article-title":"Discovery of chemical inhibitors of the selective transfer of lipids mediated by the HDL receptor SR-BI","volume":"99","author":"Nieland","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1016\/j.bbalip.2012.12.006","article-title":"Chronic pharmacological activation of P2Y13 receptor in mice decreases HDL-cholesterol level by increasing hepatic HDL uptake and bile acid secretion","volume":"1831","author":"Serhan","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"832","DOI":"10.1016\/j.bbalip.2017.05.005","article-title":"Ectopic adenine nucleotide translocase activity controls extracellular ADP levels and regulates the F1-ATPase-mediated HDL endocytosis pathway on hepatocytes","volume":"1862","author":"Cardouat","year":"2017","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1161\/ATVBAHA.111.238063","article-title":"The beta-chain of cell surface F(0)F(1) ATPase modulates apoA-I and HDL transcytosis through aortic endothelial cells","volume":"32","author":"Cavelier","year":"2012","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.amjcard.2013.09.006","article-title":"High-density lipoprotein mediated cellular cholesterol efflux in acute coronary syndromes","volume":"113","author":"Hafiane","year":"2014","journal-title":"Am. J. Cardiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1126\/science.3513311","article-title":"A receptor-mediated pathway for cholesterol homeostasis","volume":"232","author":"Brown","year":"1986","journal-title":"Science"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.1177\/000331979004101207","article-title":"High density lipoprotein inhibits low density lipoprotein binding and uptake by bovine aortic endothelial cells","volume":"41","author":"Alexander","year":"1990","journal-title":"Angiology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0022-4804(90)90128-O","article-title":"Competitive inhibition of LDL binding and uptake by HDL in aortic endothelial cells","volume":"49","author":"Alexander","year":"1990","journal-title":"J. Surg. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1254","DOI":"10.1016\/S0021-9258(18)33186-7","article-title":"Reconstitution of apolipoprotein A-I from human high density lipoprotein with bovine brain sphingomyelin","volume":"258","author":"Swaney","year":"1983","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1843","DOI":"10.1161\/ATVBAHA.107.145672","article-title":"Preferential sphingosine-1-phosphate enrichment and sphingomyelin depletion are key features of small dense HDL3 particles: Relevance to antiapoptotic and antioxidative activities","volume":"27","author":"Kontush","year":"2007","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1161\/ATVBAHA.111.244186","article-title":"Intercellular adhesion molecule 1 engagement modulates sphingomyelinase and ceramide, supporting uptake of drug carriers by the vascular endothelium","volume":"32","author":"Serrano","year":"2012","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1161\/01.ATV.21.5.810","article-title":"Cross-linking of brain endothelial intercellular adhesion molecule (ICAM)-1 induces association of ICAM-1 with detergent-insoluble cytoskeletal fraction","volume":"21","author":"Amos","year":"2001","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1186\/1476-511X-10-224","article-title":"The apolipoprotein A-I mimetic peptide, ETC-642, reduces chronic vascular inflammation in the rabbit","volume":"10","author":"Vanags","year":"2011","journal-title":"Lipids Health Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1161\/ATVBAHA.117.309287","article-title":"Impairment of the Ability of HDL From Patients With Metabolic Syndrome but Without Diabetes Mellitus to Activate eNOS: Correction by S1P Enrichment","volume":"37","author":"Denimal","year":"2017","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1478","DOI":"10.1016\/S0006-3495(00)76399-5","article-title":"Different sphingolipids show differential partitioning into sphingolipid\/cholesterol-rich domains in lipid bilayers","volume":"79","author":"Wang","year":"2000","journal-title":"Biophys. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1038\/nrcardio.2015.124","article-title":"Dysfunctional HDL and atherosclerotic cardiovascular disease","volume":"13","author":"Rosenson","year":"2016","journal-title":"Nat. Rev. Cardiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1111\/1523-1747.ep12613748","article-title":"HMEC-1: Establishment of an immortalized human microvascular endothelial cell line","volume":"99","author":"Ades","year":"1992","journal-title":"J. Investig. Dermatol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"174","DOI":"10.2174\/187152807781696428","article-title":"Dehydroepiandrosterone delays LDL oxidation in vitro and attenuates several oxLDL-induced inflammatory responses in endothelial cells","volume":"6","author":"Zentella","year":"2007","journal-title":"Inflamm. Allergy Drug Targets"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1016\/0076-6879(86)28092-1","article-title":"Reconstitution of high-density lipoproteins","volume":"128","author":"Jonas","year":"1986","journal-title":"Methods Enzymol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1081\/ERC-200035433","article-title":"Contribution of cholesteryl ester transfer protein and lecithin:cholesterol acyltransferase to HDL size distribution","volume":"30","author":"Mejia","year":"2004","journal-title":"Endocr. Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/11\/3394\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:26:55Z","timestamp":1760196415000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/11\/3394"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10,30]]},"references-count":39,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2018,11]]}},"alternative-id":["ijms19113394"],"URL":"https:\/\/doi.org\/10.3390\/ijms19113394","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,10,30]]}}}